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Huang P, Yang Y, Lian J, Yu T, Li G, Zhang Y. Neutrophils disrupt the intestinal barrier via IL-22/TGF-β/Mmp9 axis in the zebrafish model of inflammatory bowel disease. J Genet Genomics 2025:S1673-8527(25)00121-3. [PMID: 40288520 DOI: 10.1016/j.jgg.2025.04.012] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2024] [Revised: 04/17/2025] [Accepted: 04/17/2025] [Indexed: 04/29/2025]
Affiliation(s)
- Peixian Huang
- Innovation Centre of Ministry of Education for Development and Diseases, School of Medicine, South China University of Technology, Guangzhou 510006, China; Department of Intensive Care Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), School of Medicine, South China University of Technology, Guangzhou 510080, China
| | - Yiqing Yang
- Innovation Centre of Ministry of Education for Development and Diseases, School of Medicine, South China University of Technology, Guangzhou 510006, China
| | - Junwei Lian
- The Sixth Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510006, China
| | - Tao Yu
- Biomedical Research Institute, Shenzhen Peking University-the Hong Kong University of Science and Technology Medical Center, Shenzhen 518036, China; Greater Bay Biomedical Innocenter, Shenzhen Bay Laboratory, Shenzhen 518055, China.
| | - Gaofei Li
- Innovation Centre of Ministry of Education for Development and Diseases, School of Medicine, South China University of Technology, Guangzhou 510006, China; Greater Bay Biomedical Innocenter, Shenzhen Bay Laboratory, Shenzhen 518055, China.
| | - Yiyue Zhang
- Innovation Centre of Ministry of Education for Development and Diseases, School of Medicine, South China University of Technology, Guangzhou 510006, China; The Sixth Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510006, China.
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Huang X, Cai M, Yan L, Xiao R, Mu Y, Ren Y. Assessment of air pollutant O 3 pulmonary exposure using a bronchus-on-chip model coupling with atmospheric simulation chamber. JOURNAL OF HAZARDOUS MATERIALS 2025; 486:137106. [PMID: 39764952 DOI: 10.1016/j.jhazmat.2025.137106] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/30/2024] [Revised: 12/26/2024] [Accepted: 01/01/2025] [Indexed: 03/12/2025]
Abstract
Heavy air pollution is now a serious public health issue. Many studies have shown strong connections between ozone (O3) with the occurrence and development of various respiratory diseases. However, the exact mechanism is still a matter of debate. In this work, we developed a human bronchial epithelial cells (HBECs) chip that differentiates different functional cell groups of ciliated, goblet, and club cells to model the pulmonary bronchial barrier function. Concurrently, we designed an Atmospheric-Biochemical-Chip reactor (ABC-reactor), a system that could simulate different levels of O3 and particle matter. Coupling the HBECs-on-chip model with ABC-reactor, we investigated the effects of O3 at 400 ppbv and 200 ppbv on the pulmonary bronchial barrier. Our results showed that O3 at 400 ppbv severely disrupted the bronchial barrier and upregulated the expression of pro-inflammatory cytokines. However, 200 ppbv of O3 did not cause severe barrier impairment but induced cellular dysfunction, apoptosis, and reduced immune response. These suggest that bronchial trauma does exist at 200 ppbv of O3 but is not easily detected by the body due to the reduced inflammatory response. However, more research is needed to understand if the trauma induced by 200 ppbv of O3 is reversible and the interaction mechanism between O3 and PM2.5.
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Affiliation(s)
- Xuanming Huang
- Laboratory of Atmospheric Environment and Pollution Control (LAEPC), Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049, China
| | - Min Cai
- Laboratory of Atmospheric Environment and Pollution Control (LAEPC), Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China
| | - Lei Yan
- Beijing Daxiang Biotech Co., Ltd, Beijing 100191, China
| | - Rongrong Xiao
- Beijing Daxiang Biotech Co., Ltd, Beijing 100191, China
| | - Yujing Mu
- Laboratory of Atmospheric Environment and Pollution Control (LAEPC), Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049, China
| | - Yangang Ren
- Laboratory of Atmospheric Environment and Pollution Control (LAEPC), Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China; College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049, China.
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Hirano S, Aoki K, Ueno N. Dynamic behavior of cell-cell adhesion factors in collective cell migration. Cells Dev 2025:203995. [PMID: 39862903 DOI: 10.1016/j.cdev.2025.203995] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/08/2024] [Revised: 01/20/2025] [Accepted: 01/21/2025] [Indexed: 01/27/2025]
Abstract
Collective cell migration is a fundamental process underlying various biological phenomena, including embryonic development and cancer cell invasion. The cohesive yet flexible movement of cell collectives largely depends on the coordinated regulation of cell-cell and cell-substrate adhesions. In this review, we summarize the regulation of key cell-cell junction components, such as cadherins and zonula occludens proteins during collective cell migration, with a particular focus on the recently discovered multifaceted roles of ZO-1 in both cell-cell and cell-substrate interactions.
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Affiliation(s)
- Sayuki Hirano
- Laboratory of Cell Cycle Regulation, Department of Gene Mechanisms, Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Japan; Quantitative Biology Research Group, Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Japan; Division of Quantitative Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, Japan; Quantitative and Imaging Biology, International Research Collaboration Center (IRCC), National Institutes of Natural Sciences (NINS), Japan.
| | - Kazuhiro Aoki
- Laboratory of Cell Cycle Regulation, Department of Gene Mechanisms, Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Japan; Quantitative Biology Research Group, Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Japan; Division of Quantitative Biology, National Institute for Basic Biology, National Institutes of Natural Sciences, Japan; Quantitative and Imaging Biology, International Research Collaboration Center (IRCC), National Institutes of Natural Sciences (NINS), Japan; Center for Living Systems Information Science, Graduate School of Biostudies, Kyoto University, Japan
| | - Naoto Ueno
- Quantitative and Imaging Biology, International Research Collaboration Center (IRCC), National Institutes of Natural Sciences (NINS), Japan; Trans-Scale Biology Center, National Institute for Basic Biology (NIBB), National Institutes of Natural Sciences (NINS), Japan.
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4
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Ruan Y, Huang X, Sun P, Yu X, Tan X, Song Y, Chen H, Liu Z. ZO-1 boosts the in vitro self-renewal of pre-haematopoietic stem cells from OCT4-reprogrammed human hair follicle mesenchymal stem cells through cytoskeleton remodeling. Stem Cell Res Ther 2024; 15:480. [PMID: 39696518 PMCID: PMC11658245 DOI: 10.1186/s13287-024-04080-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/12/2024] [Accepted: 11/25/2024] [Indexed: 12/20/2024] Open
Abstract
BACKGROUND The challenge of expanding haematopoietic stem/progenitor cells (HSPCs) in vitro has limited their clinical application. Human hair follicle mesenchymal stem cells (hHFMSCs) can be reprogrammed to generate intermediate stem cells by transducing OCT4 (hHFMSCsOCT4) and pre-inducing with FLT3LG/SCF, and differentiated into erythrocytes. These intermediate cells exhibit gene expression patterns similar to pre-HSCs, making them promising for artificial haematopoiesis. However, further investigation is required to elucidate the in vitro proliferation ability and mechanism underlying the self-renewal of pre-HSCs derived from hHFMSCs. METHODS hHFMSCsOCT4 were pre-treated with FLT3LG and SCF cytokines, followed by characterization and isolation of the floating cell subsets for erythroid differentiation through stimulation with hematopoietic cytokines and nutritional factors. Cell adhesion was assessed through disassociation and adhesion assays. OCT4 expression levels were measured using immunofluorescence staining, RT-qPCR, and Western blotting. RNA sequencing and Gene Ontology (GO) enrichment analysis were then conducted to identify proliferation-related biological processes. Proliferative capacity was evaluated through CCK-8, colony formation assays, Ki67 index, and cell cycle analysis. Cytoskeleton was observed through Wright‒Giemsa, Coomassie brilliant blue, and phalloidin staining. Expression of adherens junction (AJ) core members was confirmed through RT‒qPCR, Western blotting, and immunofluorescence staining before and after ZO-1 knockdown. A regulatory network was constructed to determine relationships among cytoskeleton, proliferation, and the AJ pathway. Student's t tests (GraphPad Prism 8.0.2) were used for group comparisons. The results were considered significant at P < 0.05. RESULTS Pre-treatment of hHFMSCsOCT4 with FLT3LG and SCF leads to the emergence of floating cell subsets exhibiting small, globoid morphology, suspended above adherent cells, forming colonies, and displaying minimal expression of CD45. Excessive OCT4 expression weakens adhesion in floating hHFMSCsOCT4. Floating cells moderately enhanced proliferation and undergo cytoskeleton remodelling, with increased contraction and aggregation of F-actin near the nucleus. The upregulation of ZO-1 could impact the expressions of F-actin, E-cadherin, and β-catenin genes, as well as the nuclear positioning of β-catenin, leading to variations in the cytoskeleton and cell cycle. Finally, a regulatory network revealed that the AJ pathway cored with ZO-1 critically bridges cytoskeletal remodelling and haematopoiesis-related proliferation in a β-catenin-dependent manner. CONCLUSIONS ZO-1 improved the self-renewal of pre-HSCs from OCT4-overexpressing hHFMSCs by remodeling the cytoskeleton via the ZO-1-regulated AJ pathway, suggesting floating hHFMSCsOCT4 as the promising seed cells for artificial hematopoiesis.
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Affiliation(s)
- Yingchun Ruan
- Department of Pathology, Qingdao Municipal Hospital Group, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China
| | - Xingang Huang
- Department of Pathology, Qingdao Municipal Hospital Group, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China
| | - Pengpeng Sun
- Department of Critical Care Medicine, Qingdao Central Hospital, University of Health and Rehabilitation Sciences (Qingdao Central Hospital), 127 Siliunan Road, Qingdao, 266042, Shandong, China
| | - Xiaozhen Yu
- Department of Pathology, Qingdao Municipal Hospital Group, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China
| | - Xiaohua Tan
- Department of Pathology, College of Basic Medical Sciences, Qingdao University, 308 Ningxia Road, Qingdao, 266071, Shandong, China
| | - Yaolin Song
- Department of Pathology, The Affiliated Hospital of Qingdao University, 16 Jiangsu Road, Qingdao, 266000, Shandong, China
| | - Hua Chen
- Department of Pathology, Qingdao Municipal Hospital Group, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China
| | - Zhijing Liu
- Department of Pathology, Qingdao Municipal Hospital Group, 1 Jiaozhou Road, Qingdao, 266011, Shandong, China.
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Oikonomou I, Papageorgiou A, de Lastic AL, Moulias A, Georgopoulou GA, Mouzaki A, Koufou EE, Tsigkas G, Gogos C, Davlouros P, Assimakopoulos SF. Gut barrier dysfunction, endotoxemia and inflammatory response in STEMI patients and effect of primary PCI. Am J Med Sci 2024; 368:485-493. [PMID: 38969287 DOI: 10.1016/j.amjms.2024.07.004] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/22/2023] [Revised: 06/05/2024] [Accepted: 07/01/2024] [Indexed: 07/07/2024]
Abstract
BACKGROUND Gut-derived bacterial and endotoxin translocation induce systemic inflammation, which exerts a pivotal pathogenetic role in all phases of atherosclerosis. OBJECTIVES To investigate prospectively the gut barrier function, endotoxin translocation and inflammatory response in ST-elevation myocardial infarction (STEMI) patients undergoing primary percutaneous coronary artery intervention (PPCI). METHODS Twenty-seven patients with STEMI that underwent successful PPCI were subjected to peripheral blood sampling at 3-time points; before PPCI (day0), 24 h (day1) and 96 h (day4) after PPCI and were compared with 20 chronic coronary syndrome (CCS) patients and 11 healthy controls. Serum ZO-1, I-FABP and endotoxin concentrations were determined by ELISA. Concentrations of cytokines IL-1β, -6, -8, -10 and TNF-α were determined by flow cytometry. RESULTS Patients with STEMI before PPCI (day0) had increased serum ZO-1 and endotoxin, both at significantly higher levels compared to CCS patients. STEMI induced also significant increases of the cytokines IL-6, -8 and -10. After PPCI, a significant improvement of gut barrier integrity (ZO-1) and endotoxemia was observed from the first day. At day4 post PPCI, systemic endotoxin and cytokines IL-6, -8 and -10 levels were reduced to control levels. Serum ZO-1 levels were positively correlated with systemic IL-10 concentrations (r = 0.471). CONCLUSION STEMI is associated with gut barrier dysfunction, systemic endotoxemia and inflammatory response, which improve rapidly following successful PPCI.
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Affiliation(s)
- Ioanna Oikonomou
- Department of Internal Medicine and Division of Infectious Diseases, University of Patras Medical School, Patras, Greece
| | - Angeliki Papageorgiou
- Division of Cardiology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Anne-Lise de Lastic
- Laboratory of Immunohematology, Division of Hematology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Athanasios Moulias
- Division of Cardiology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | | | - Athanasia Mouzaki
- Laboratory of Immunohematology, Division of Hematology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Eleni-Evangelia Koufou
- Division of Cardiology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Grigorios Tsigkas
- Division of Cardiology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Charalambos Gogos
- Department of Internal Medicine and Division of Infectious Diseases, University of Patras Medical School, Patras, Greece
| | - Periklis Davlouros
- Division of Cardiology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Stelios F Assimakopoulos
- Department of Internal Medicine and Division of Infectious Diseases, University of Patras Medical School, Patras, Greece.
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Yan Q, Wang Q, Zhang Y, Yuan L, Hu J, Zhao X. The Novel-m0230-3p miRNA Modulates the CSF1/CSF1R/Ras Pathway to Regulate the Cell Tight Junctions and Blood-Testis Barrier in Yak. Cells 2024; 13:1304. [PMID: 39120333 PMCID: PMC11311379 DOI: 10.3390/cells13151304] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/25/2024] [Revised: 08/02/2024] [Accepted: 08/02/2024] [Indexed: 08/10/2024] Open
Abstract
The yak (Bos grunniens) is a valuable livestock animal endemic to the Qinghai-Tibet Plateau in China with low reproductive rates. Cryptorchidism is one of the primary causes of infertility in male yaks. Compared with normal testes, the tight junctions (TJs) of Sertoli cells (SCs) and the integrity of the blood-testis barrier (BTB) in cryptorchidism are both disrupted. MicroRNAs are hairpin-derived RNAs of about 19-25 nucleotides in length and are involved in a variety of biological processes. Numerous studies have shown the involvement of microRNAs in the reproductive physiology of yak. In this study, we executed RNA sequencing (RNA-seq) to describe the expression profiles of mRNAs and microRNAs in yaks with normal testes and cryptorchidism to identify differentially expressed genes. GO and KEGG analyses were used to identify the biological processes and signaling pathways which the target genes of the differentially expressed microRNAs primarily engaged. It was found that novel-m0230-3p is an important miRNA that significantly differentiates between cryptorchidism and normal testes, and it is down-regulated in cryptorchidism with p < 0.05. Novel-m0230-3p and its target gene CSF1 both significantly contribute to the regulation of cell adhesion and tight junctions. The binding sites of novel-m0230-3p with CSF1 were validated by a dual luciferase reporter system. Then, mimics and inhibitors of novel-m0230-3p were transfected in vitro into SCs, respectively. A further analysis using qRT-PCR, immunofluorescence (IF), and Western blotting confirmed that the expression of cell adhesion and tight-junction-related proteins Occludin and ZO-1 both showed changes. Specifically, both the mRNA and protein expression levels of Occludin and ZO-1 in SCs decreased after transfection with the novel-m0230-3p mimics, while they increased after transfection with the inhibitors, with p < 0.05. These were achieved via the CSF1/CSF1R/Ras signaling pathway. In summary, our findings indicate a negative miRNA-mRNA regulatory network involving the CSF1/CSF1R/Ras signaling pathway in yak SCs. These results provide new insights into the molecular mechanisms of CSF1 and suggest that novel-m0230-3p and its target protein CSF1 could be used as potential therapeutic targets for yak cryptorchidism.
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Affiliation(s)
- Qiu Yan
- College of Veterinary Medicine, Gansu Agriculture University, Lanzhou 730070, China; (Y.Z.); (L.Y.); (J.H.); (X.Z.)
- Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
| | - Qi Wang
- College of Veterinary Medicine, Gansu Agriculture University, Lanzhou 730070, China; (Y.Z.); (L.Y.); (J.H.); (X.Z.)
- Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
| | - Yong Zhang
- College of Veterinary Medicine, Gansu Agriculture University, Lanzhou 730070, China; (Y.Z.); (L.Y.); (J.H.); (X.Z.)
- Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
| | - Ligang Yuan
- College of Veterinary Medicine, Gansu Agriculture University, Lanzhou 730070, China; (Y.Z.); (L.Y.); (J.H.); (X.Z.)
- Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
| | - Junjie Hu
- College of Veterinary Medicine, Gansu Agriculture University, Lanzhou 730070, China; (Y.Z.); (L.Y.); (J.H.); (X.Z.)
- Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
| | - Xingxu Zhao
- College of Veterinary Medicine, Gansu Agriculture University, Lanzhou 730070, China; (Y.Z.); (L.Y.); (J.H.); (X.Z.)
- Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou 730070, China
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Thomas D, Recabal-Beyer A, Senecal JMM, Serletis D, Lynn BD, Jackson MF, Nagy JI. Association of connexin36 with adherens junctions at mixed synapses and distinguishing electrophysiological features of those at mossy fiber terminals in rat ventral hippocampus. INTERNATIONAL JOURNAL OF PHYSIOLOGY, PATHOPHYSIOLOGY AND PHARMACOLOGY 2024; 16:28-54. [PMID: 39021415 PMCID: PMC11249852 DOI: 10.62347/rtmh4490] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/28/2024] [Accepted: 05/06/2024] [Indexed: 07/20/2024]
Abstract
BACKGROUND Granule cells in the hippocampus project axons to hippocampal CA3 pyramidal cells where they form large mossy fiber terminals. We have reported that these terminals contain the gap junction protein connexin36 (Cx36) specifically in the stratum lucidum of rat ventral hippocampus, thus creating morphologically mixed synapses that have the potential for dual chemical/electrical transmission. METHODOLOGY Here, we used various approaches to characterize molecular and electrophysiological relationships between the Cx36-containing gap junctions at mossy fiber terminals and their postsynaptic elements and to examine molecular relationships at mixed synapses in the brainstem. RESULTS In rat and human ventral hippocampus, many of these terminals, identified by their selective expression of vesicular zinc transporter-3 (ZnT3), displayed multiple, immunofluorescent Cx36-puncta representing gap junctions, which were absent at mossy fiber terminals in the dorsal hippocampus. In rat, these were found in close proximity to the protein constituents of adherens junctions (i.e., N-cadherin and nectin-1) that are structural hallmarks of mossy fiber terminals, linking these terminals to the dendritic shafts of CA3 pyramidal cells, thus indicating the loci of gap junctions at these contacts. Cx36-puncta were also associated with adherens junctions at mixed synapses in the brainstem, supporting emerging views of the structural organization of the adherens junction-neuronal gap junction complex. Electrophysiologically induced long-term potentiation (LTP) of field responses evoked by mossy fiber stimulation was greater in the ventral than dorsal hippocampus. CONCLUSIONS The electrical component of transmission at mossy fiber terminals may contribute to enhanced LTP responses in the ventral hippocampus.
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Affiliation(s)
- Deepthi Thomas
- Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of ManitobaWinnipeg, Manitoba, Canada
| | - Antonia Recabal-Beyer
- Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of ManitobaWinnipeg, Manitoba, Canada
- Departamento de Biología Celular, Facultad de Ciencias Biológicas, Universidad de ConcepciónVíctor Lamas 1290, Casilla 160, Concepción, Chile
| | - Joanne MM Senecal
- Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of ManitobaWinnipeg, Manitoba, Canada
| | - Demitre Serletis
- Epilepsy Center, Neurological Institute, Cleveland ClinicCleveland, Ohio, USA
| | - Bruce D Lynn
- Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of ManitobaWinnipeg, Manitoba, Canada
| | - Michael F Jackson
- Department of Pharmacology and Therapeutics, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of ManitobaWinnipeg, Manitoba, Canada
- PrairieNeuro Research Centre, Kleysen Institute for Advanced Medicine, Health Science CentreWinnipeg, Manitoba, Canada
| | - James I Nagy
- Department of Physiology and Pathophysiology, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of ManitobaWinnipeg, Manitoba, Canada
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8
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Nguyen TP, Otani T, Tsutsumi M, Kinoshita N, Fujiwara S, Nemoto T, Fujimori T, Furuse M. Tight junction membrane proteins regulate the mechanical resistance of the apical junctional complex. J Cell Biol 2024; 223:e202307104. [PMID: 38517380 PMCID: PMC10959758 DOI: 10.1083/jcb.202307104] [Citation(s) in RCA: 9] [Impact Index Per Article: 9.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/02/2023] [Revised: 01/29/2024] [Accepted: 02/16/2024] [Indexed: 03/23/2024] Open
Abstract
Epithelia must be able to resist mechanical force to preserve tissue integrity. While intercellular junctions are known to be important for the mechanical resistance of epithelia, the roles of tight junctions (TJs) remain to be established. We previously demonstrated that epithelial cells devoid of the TJ membrane proteins claudins and JAM-A completely lack TJs and exhibit focal breakages of their apical junctions. Here, we demonstrate that apical junctions fracture when claudin/JAM-A-deficient cells undergo spontaneous cell stretching. The junction fracture was accompanied by actin disorganization, and actin polymerization was required for apical junction integrity in the claudin/JAM-A-deficient cells. Further deletion of CAR resulted in the disruption of ZO-1 molecule ordering at cell junctions, accompanied by severe defects in apical junction integrity. These results demonstrate that TJ membrane proteins regulate the mechanical resistance of the apical junctional complex in epithelial cells.
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Affiliation(s)
- Thanh Phuong Nguyen
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Japan
- Physiological Sciences Program, Graduate Institute for Advanced Studies, SOKENDAI, Okazaki, Japan
| | - Tetsuhisa Otani
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Japan
- Physiological Sciences Program, Graduate Institute for Advanced Studies, SOKENDAI, Okazaki, Japan
- Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Kawaguchi, Japan
| | - Motosuke Tsutsumi
- Division of Biophotonics, National Institute for Physiological Sciences, Okazaki, Japan
- Biophotonics Research Group, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Okazaki, Japan
| | - Noriyuki Kinoshita
- Division of Embryology, National Institute for Basic Biology, Okazaki, Japan
- Basic Biology Program, Graduate Institute for Advanced Studies, SOKENDAI, Okazaki, Japan
| | - Sachiko Fujiwara
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Japan
- Physiological Sciences Program, Graduate Institute for Advanced Studies, SOKENDAI, Okazaki, Japan
| | - Tomomi Nemoto
- Physiological Sciences Program, Graduate Institute for Advanced Studies, SOKENDAI, Okazaki, Japan
- Division of Biophotonics, National Institute for Physiological Sciences, Okazaki, Japan
- Biophotonics Research Group, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Okazaki, Japan
| | - Toshihiko Fujimori
- Division of Embryology, National Institute for Basic Biology, Okazaki, Japan
- Basic Biology Program, Graduate Institute for Advanced Studies, SOKENDAI, Okazaki, Japan
| | - Mikio Furuse
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Japan
- Physiological Sciences Program, Graduate Institute for Advanced Studies, SOKENDAI, Okazaki, Japan
- Nagoya University Graduate School of Medicine, Nagoya, Japan
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9
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Casanova-Sepúlveda G, Sexton JA, Turk BE, Boggon TJ. Autoregulation of the LIM kinases by their PDZ domain. Nat Commun 2023; 14:8441. [PMID: 38114480 PMCID: PMC10730565 DOI: 10.1038/s41467-023-44148-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/15/2023] [Accepted: 12/01/2023] [Indexed: 12/21/2023] Open
Abstract
LIM domain kinases (LIMK) are important regulators of actin cytoskeletal remodeling. These protein kinases phosphorylate the actin depolymerizing factor cofilin to suppress filament severing, and are key nodes between Rho GTPase cascades and actin. The two mammalian LIMKs, LIMK1 and LIMK2, contain consecutive LIM domains and a PDZ domain upstream of the C-terminal kinase domain. The roles of the N-terminal regions are not fully understood, and the function of the PDZ domain remains elusive. Here, we determine the 2.0 Å crystal structure of the PDZ domain of LIMK2 and reveal features not previously observed in PDZ domains including a core-facing arginine residue located at the second position of the 'x-Φ-G-Φ' motif, and that the expected peptide binding cleft is shallow and poorly conserved. We find a distal extended surface to be highly conserved, and when LIMK1 was ectopically expressed in yeast we find targeted mutagenesis of this surface decreases growth, implying increased LIMK activity. PDZ domain LIMK1 mutants expressed in yeast are hyperphosphorylated and show elevated activity in vitro. This surface in both LIMK1 and LIMK2 is critical for autoregulation independent of activation loop phosphorylation. Overall, our study demonstrates the functional importance of the PDZ domain to autoregulation of LIMKs.
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Affiliation(s)
| | - Joel A Sexton
- Department of Pharmacology, Yale University, New Haven, CT, 06520, USA
| | - Benjamin E Turk
- Department of Pharmacology, Yale University, New Haven, CT, 06520, USA
| | - Titus J Boggon
- Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, 06520, USA.
- Department of Pharmacology, Yale University, New Haven, CT, 06520, USA.
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10
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Fan M, Deng F, Tang R, Cai Y, Zhang X, Li H, Xiang T, Pan J. Serum Zonula Occludens-1 and Claudin-5 Levels in Patients with Insomnia Disorder: A Pilot Study. Nat Sci Sleep 2023; 15:873-884. [PMID: 37928369 PMCID: PMC10625320 DOI: 10.2147/nss.s424756] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/07/2023] [Accepted: 10/18/2023] [Indexed: 11/07/2023] Open
Abstract
Purpose This research aimed to investigate serum Zonula occludens-1 (ZO-1) and Claudin-5 (CLDN5) levels to show whether or not their eventual changes in patients with insomnia disorder could have etiopathogenetic importance. There was no research investigating serum ZO-1 and CLDN5 concentrations in insomnia disorder. Patients and Methods This study included 60 insomnia disorder patients and 45 normal controls. None of the patients received drugs for insomnia. The patients completed Insomnia Severity Index (ISI) and Pittsburgh Sleep Quality Index (PSQI), and Polysomnography (PSG) to score the insomnia disorder symptoms. Venous blood samples were collected, and serum ZO-1 and claudin-5 levels were analyzed by enzyme-linked immunosorbent assay (ELISA). Results Serum ZO-1 level was significantly higher without a significant difference between age, sex, and body mass index, whereas the difference in serum claudin-5 level between the two groups was not statistically significant. In addition, ZO-1 levels were positively correlated with ISI and PSQI and negatively with N1 and N1_perc. We also demonstrated a positive correlation between the levels of CLDN5 and HAMA, and a negative correlation with total sleep time (TST), N1 and N1_perc. Conclusion Our findings suggest an association between these intestinal and brain endothelial permeability markers and insomnia disorders. However, these remain modest and preliminary and need more extensive studies, including long-term follow-up populations and involving gut microbes, to further validate and explore the mechanisms involved.
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Affiliation(s)
- Mei Fan
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
| | - Fangyi Deng
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
| | - Rui Tang
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
| | - Yixian Cai
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
| | - Xiaotao Zhang
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
| | - Hongyao Li
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
| | - Ting Xiang
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
| | - Jiyang Pan
- Department of Psychiatry, Sleep Medicine Centre, The First Affiliated Hospital, Jinan University, Guangzhou, People's Republic of China
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11
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Jiang X, Xu Z, Jiang S, Wang H, Xiao M, Shi Y, Wang K. PDZ and LIM Domain-Encoding Genes: Their Role in Cancer Development. Cancers (Basel) 2023; 15:5042. [PMID: 37894409 PMCID: PMC10605254 DOI: 10.3390/cancers15205042] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2023] [Revised: 10/13/2023] [Accepted: 10/15/2023] [Indexed: 10/29/2023] Open
Abstract
PDZ-LIM family proteins (PDLIMs) are a kind of scaffolding proteins that contain PDZ and LIM interaction domains. As protein-protein interacting molecules, PDZ and LIM domains function as scaffolds to bind to a variety of proteins. The PDLIMs are composed of evolutionarily conserved proteins found throughout different species. They can participate in cell signal transduction by mediating the interaction of signal molecules. They are involved in many important physiological processes, such as cell differentiation, proliferation, migration, and the maintenance of cellular structural integrity. Studies have shown that dysregulation of the PDLIMs leads to tumor formation and development. In this paper, we review and integrate the current knowledge on PDLIMs. The structure and function of the PDZ and LIM structural domains and the role of the PDLIMs in tumor development are described.
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Affiliation(s)
| | | | | | | | | | - Yueli Shi
- Department of Respiratory and Critical Care Medicine, The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu 322000, China; (X.J.); (Z.X.); (S.J.); (H.W.); (M.X.)
| | - Kai Wang
- Department of Respiratory and Critical Care Medicine, The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu 322000, China; (X.J.); (Z.X.); (S.J.); (H.W.); (M.X.)
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12
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Castro-Cruz M, Lembo F, Borg JP, Travé G, Vincentelli R, Zimmermann P. The Human PDZome 2.0: Characterization of a New Resource to Test for PDZ Interactions by Yeast Two-Hybrid. MEMBRANES 2023; 13:737. [PMID: 37623798 PMCID: PMC10456741 DOI: 10.3390/membranes13080737] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/04/2023] [Revised: 07/28/2023] [Accepted: 08/10/2023] [Indexed: 08/26/2023]
Abstract
PSD95-disc large-zonula occludens (PDZ) domains are globular modules of 80-90 amino acids that co-evolved with multicellularity. They commonly bind to carboxy-terminal sequences of a plethora of membrane-associated proteins and influence their trafficking and signaling. We previously built a PDZ resource (PDZome) allowing us to unveil human PDZ interactions by Yeast two-hybrid. Yet, this resource is incomplete according to the current knowledge on the human PDZ proteome. Here we built the PDZome 2.0 library for Yeast two-hybrid, based on a PDZ library manually curated from online resources. The PDZome2.0 contains 305 individual clones (266 PDZ domains in isolation and 39 tandems), for which all boundaries were designed based on available PDZ structures. Using as bait the E6 oncoprotein from HPV16, a known promiscuous PDZ interactor, we show that PDZome 2.0 outperforms the previous resource.
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Affiliation(s)
- Monica Castro-Cruz
- Department of Human Genetics, KU Leuven, 3000 Leuven, Belgium;
- Équipe Labellisée Ligue 2018, Centre de Recherche en Cancérologie de Marseille (CRCM), Aix-Marseille Université, 13009 Marseille, France;
| | - Frédérique Lembo
- Équipe Labellisée Ligue 2018, Centre de Recherche en Cancérologie de Marseille (CRCM), Aix-Marseille Université, 13009 Marseille, France;
| | - Jean-Paul Borg
- Marseille Proteomics Platform, CRCM, Institute Paoli-Calmettes, Aix-Marseille Université, Inserm, CNRS, 13009 Marseille, France;
| | - Gilles Travé
- Équipe Labellisée Ligue 2015, Department of Integrated Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), INSERM U1258/CNRS UMR 7104/Université de Strasbourg, 67404 Illkirch, France;
| | - Renaud Vincentelli
- Architecture et Fonction des Macromolécules Biologiques (AFMB), Unité Mixte de Recherche (UMR) 7257, Centre National de la Recherche Scientifique (CNRS), Aix-Marseille Université, 13009 Marseille, France;
| | - Pascale Zimmermann
- Department of Human Genetics, KU Leuven, 3000 Leuven, Belgium;
- Équipe Labellisée Ligue 2018, Centre de Recherche en Cancérologie de Marseille (CRCM), Aix-Marseille Université, 13009 Marseille, France;
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13
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Yu S, He J, Xie K. Zonula Occludens Proteins Signaling in Inflammation and Tumorigenesis. Int J Biol Sci 2023; 19:3804-3815. [PMID: 37564207 PMCID: PMC10411466 DOI: 10.7150/ijbs.85765] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/01/2023] [Accepted: 07/10/2023] [Indexed: 08/12/2023] Open
Abstract
Tight junction (TJ) is the barrier of epithelial and endothelial cells to maintain paracellular substrate transport and cell polarity. As one of the TJ cytoplasmic adaptor proteins adjacent to cell membrane, zonula occludens (ZO) proteins are responsible for connecting transmembrane TJ proteins and cytoplasmic cytoskeleton, providing a binding platform for transmembrane TJ proteins to maintain the barrier function. In addition to the basic structural function, ZO proteins play important roles in signal regulation such as cell proliferation and motility, the latter including cell migration, invasion and metastasis, to influence embryonic development, tissue homeostasis, damage repair, inflammation, tumorigenesis, and cancer progression. In this review, we will focus on the signal regulating function of ZO proteins in inflammation and tumorigenesis, and discuss the limitations of previous research and future challenges in ZO protein research.
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Affiliation(s)
- Sen Yu
- Center for Pancreatic Cancer Research, The South China University of Technology School of Medicine, Guangzhou, China
| | - Jie He
- The Second Affiliated Hospital and Guangzhou First People's Hospital, South China University of Technology School of Medicine, Guangdong, China
| | - Keping Xie
- Center for Pancreatic Cancer Research, The South China University of Technology School of Medicine, Guangzhou, China
- The Second Affiliated Hospital and Guangzhou First People's Hospital, South China University of Technology School of Medicine, Guangdong, China
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14
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Tsukita K, Kitamata M, Kashihara H, Yano T, Fujiwara I, Day TF, Katsuno T, Kim J, Takenaga F, Tanaka H, Park S, Miyata M, Watanabe H, Kondoh G, Takahashi R, Tamura A, Tsukita S. Phase separation of an actin nucleator by junctional microtubules regulates epithelial function. SCIENCE ADVANCES 2023; 9:eadf6358. [PMID: 36791197 PMCID: PMC9931218 DOI: 10.1126/sciadv.adf6358] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/03/2022] [Accepted: 01/18/2023] [Indexed: 06/18/2023]
Abstract
Liquid-liquid phase separation (LLPS) is involved in various dynamic biological phenomena. In epithelial cells, dynamic regulation of junctional actin filaments tethered to the apical junctional complex (AJC) is critical for maintaining internal homeostasis against external perturbations; however, the role of LLPS in this process remains unknown. Here, after identifying a multifunctional actin nucleator, cordon bleu (Cobl), as an AJC-enriched microtubule-associated protein, we conducted comprehensive in vitro and in vivo analyses. We found that apical microtubules promoted LLPS of Cobl at the AJC, and Cobl actin assembly activity increased upon LLPS. Thus, microtubules spatiotemporally regulated junctional actin assembly for epithelial morphogenesis and paracellular barriers. Collectively, these findings established that LLPS of the actin nucleator Cobl mediated dynamic microtubule-actin cross-talk in junctions, which fine-tuned the epithelial barrier.
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Affiliation(s)
- Kazuto Tsukita
- Advanced Comprehensive Research Organization, Teikyo University, Itabashi-ku, Tokyo 173-0003, Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
- Department of Neurology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
| | - Manabu Kitamata
- Advanced Comprehensive Research Organization, Teikyo University, Itabashi-ku, Tokyo 173-0003, Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
| | - Hiroka Kashihara
- Advanced Comprehensive Research Organization, Teikyo University, Itabashi-ku, Tokyo 173-0003, Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
| | - Tomoki Yano
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
- Department of Organoid Medicine, Sakaguchi Laboratory, Keio University School of Medicine, Shinjuku-ku, Tokyo 160-8582, Japan
| | - Ikuko Fujiwara
- Departments of Materials Science and Bioengineering, Nagaoka University of Technology, Nagaoka, Niigata 940-2188, Japan
- Graduate School of Science, Osaka Metropolitan University, Sumiyoshi-ku, Osaka 558-8585, Japan
| | - Timothy F. Day
- Advanced Comprehensive Research Organization, Teikyo University, Itabashi-ku, Tokyo 173-0003, Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
| | - Tatsuya Katsuno
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
- Center for Anatomical, Pathological and Forensic Medical Researches, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
| | - Jaewon Kim
- Graduate School of Mechanical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do 16419, Korea
| | - Fumiko Takenaga
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
| | - Hiroo Tanaka
- Advanced Comprehensive Research Organization, Teikyo University, Itabashi-ku, Tokyo 173-0003, Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
- Department of Pharmacology, Teikyo University School of Medicine, Itabashi-ku, Tokyo 173-8605, Japan
| | - Sungsu Park
- Graduate School of Mechanical Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do 16419, Korea
| | - Makoto Miyata
- Graduate School of Science, Osaka Metropolitan University, Sumiyoshi-ku, Osaka 558-8585, Japan
| | - Hitomi Watanabe
- Laboratory of Integrative Biological Science, Institute for Frontier Life and Medical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
| | - Gen Kondoh
- Laboratory of Integrative Biological Science, Institute for Frontier Life and Medical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
| | - Ryosuke Takahashi
- Department of Neurology, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan
| | - Atsushi Tamura
- Advanced Comprehensive Research Organization, Teikyo University, Itabashi-ku, Tokyo 173-0003, Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
- Department of Pharmacology, Teikyo University School of Medicine, Itabashi-ku, Tokyo 173-8605, Japan
| | - Sachiko Tsukita
- Advanced Comprehensive Research Organization, Teikyo University, Itabashi-ku, Tokyo 173-0003, Japan
- Laboratory of Barriology and Cell Biology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan
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15
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Chan CH, Lin P, Yang TY, Bao BY, Jhong JY, Weng YP, Lee TH, Cheng HF, Lu TL. Epithelial polarization in the 3D matrix requires MST3 signaling to regulate ZO-1 position. PLoS One 2023; 18:e0285217. [PMID: 37155619 PMCID: PMC10166550 DOI: 10.1371/journal.pone.0285217] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/09/2023] [Accepted: 04/17/2023] [Indexed: 05/10/2023] Open
Abstract
Apical-basal cell polarity must be tightly controlled for epithelial cyst and tubule formation, and these are important functional units in various epithelial organs. Polarization is achieved through the coordination of several molecules that divide cells into an apical domain and a basolateral domain, which are separated from tight and adherens junctions. Cdc42 regulates cytoskeletal organization and the tight junction protein ZO-1 at the apical margin of epithelial cell junctions. MST kinases control organ size through the regulation of cell proliferation and cell polarity. For example, MST1 relays the Rap1 signal to induce cell polarity and adhesion of lymphocytes. Our previous study showed that MST3 was involved in E-cadherin regulation and migration in MCF7 cells. In vivo, MST3 knockout mice exhibited higher ENaC expression at the apical site of renal tubules, resulting in hypertension. However, it was not clear whether MST3 was involved in cell polarity. Here, control MDCK cells, HA-MST3 and HA-MST3 kinase-dead (HA-MST3-KD) overexpressing MDCK cells were cultured in collagen or Matrigel. We found that the cysts of HA-MST3 cells were fewer and smaller than those of control MDCK cells; ZO-1 was delayed to the apical site of cysts and in cell-cell contact in the Ca2+ switch assay. However, HA-MST3-KD cells exhibited multilumen cysts. Intensive F-actin stress fibers were observed in HA-MST3 cells with higher Cdc42 activity; in contrast, HA-MST3-KD cells had lower Cdc42 activity and weaker F-actin staining. In this study, we identified a new MST3 function in the establishment of cell polarity through Cdc42 regulation.
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Affiliation(s)
- Chee-Hong Chan
- Department of Nephrology, Chang Bing Show Chwan Memorial Hospital, Lukang, Changhua, Taiwan
| | - Pei Lin
- Division of Cardiology, Department of Internal Medicine, An Nan Hospital, China Medical University, Tainan, Taiwan
| | - Tse-Yen Yang
- Molecular and Genomic Epidemiology Center, Department of Medical Research, China Medical University, Tainan, Taiwan
| | - Bo-Ying Bao
- College of School of Pharmacy, China Medical University, Tainan, Taiwan
| | - Jhen-Yang Jhong
- Department of Medical Laboratory Science and Biotechnology, Sin-Lau Hospital, Tainan, Taiwan
| | - Yui-Ping Weng
- Department of Acupressure Technology, Chung Hwa University of Medical Technology, Tainan, Taiwan
| | - Te-Hsiu Lee
- Department of Medical Laboratory Science and Biotechnology, Chung Hwa University of Medical Technology, Tainan, Taiwan
| | - Hui-Fen Cheng
- Department of Laboratory Medicine, Tainan Municipal Hospital (Managed by Show Chwan Medical Care Corporation), Tainan, Taiwan
| | - Te-Ling Lu
- College of School of Pharmacy, China Medical University, Tainan, Taiwan
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16
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Yao W, Zhang Y, Zhang W, Wen Y, Yang R, Dong J, Zhang X, Hua Y, Ji P, Wei Y. Pathological mechanism of intestinal mucosal barrier injury of large intestine dampness-heat syndrome rats and the protective effect of Yujin powder. Res Vet Sci 2022; 152:485-496. [PMID: 36156378 DOI: 10.1016/j.rvsc.2022.09.014] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/09/2021] [Revised: 06/12/2022] [Accepted: 09/16/2022] [Indexed: 11/29/2022]
Abstract
Large intestine dampness-heat syndrome (LIDHS) is frequently-occurring in the inflammatory intestinal disease of animals and human. Yujin powder (YJP) is a classical prescription for treating LIDHS. To explore the pathological mechanism of intestinal mucosal barrier injury of LIDHS and the protection of YJP, the LIDHS rat model was established through imitating the inducing conditions of LIDHS and treated with YJP. The integrity of ileal and colonic mucosa was detected through histopathological examination. The serum DAO, D-LA and ET levels were detected by ELISA. The mRNA and protein expression levels of Occludin, ZO-1 and MUC2 in ileum and colon were detected using RT-PCR and immunohistochemistry methods, respectively. The results showed that the ileal and colonic epithelium of LIDHS rats were destroyed; the serum DAO, D-LA and ET levels were significantly increased; the mRNA and protein expression levels of Occludin, ZO-1 and MUC2 in ileum and colon were all abnormally expressed. After treatment with YJP, the mucosal integrity was restored; the levels of serum DAO, D-LA and ET, mRNA and protein levels of Occludin and ZO-1 in ileum and colon and MUC2 in ileum were back-regulated; however, MUC2 level in colon was further increased. The results demonstrated that the intestinal mucosal barrier was damaged in LIDHS rats and Occludin, ZO-1 and MUC2 were abnormally expressed, and YJP could repair the intestinal mucosal barrier through up-regulating the expression of Occludin and ZO-1 in ileum and colon as well as MUC2 in colon and down-regulating MUC2 in ileum.
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Affiliation(s)
- Wanling Yao
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Yahui Zhang
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Wangdong Zhang
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Yanqiao Wen
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Rong Yang
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Jiaqi Dong
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Xiaosong Zhang
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Yongli Hua
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Peng Ji
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China
| | - Yanming Wei
- College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China.
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17
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Yamada T, Saitoh Y, Kametani K, Kamijo A, Sakamoto T, Terada N. Involvement of membrane palmitoylated protein 2 (MPP2) in the synaptic molecular complex at the mouse cerebellar glomerulus. Histochem Cell Biol 2022; 158:497-511. [PMID: 35854144 DOI: 10.1007/s00418-022-02137-6] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 06/28/2022] [Indexed: 11/30/2022]
Abstract
We previously reported that the membrane skeletal protein 4.1G in the peripheral nervous system transports membrane palmitoylated protein 6 (MPP6), which interacts with the synaptic scaffolding protein Lin7 and cell adhesion molecule 4 (CADM4) in Schwann cells that form myelin. In the present study, we investigated the localization of and proteins related to MPP2, a highly homologous family protein of MPP6, in the cerebellum of the mouse central nervous system, in which neurons are well organized. Immunostaining for MPP2 was observed at cerebellar glomeruli (CG) in the granular layer after postnatal day 14. Using the high-resolution Airyscan mode of a confocal laser-scanning microscope, MPP2 was detected as a dot pattern and colocalized with CADM1 and Lin7, recognized as small ring/line patterns, as well as with calcium/calmodulin-dependent serine protein kinase (CASK), NMDA glutamate receptor 1 (GluN1), and M-cadherin, recognized as dot patterns, indicating the localization of MPP2 in the excitatory postsynaptic region and adherens junctions of granule cells. An immunoprecipitation analysis revealed that MPP2 formed a molecular complex with CADM1, CASK, M-cadherin, and Lin7. Furthermore, the Lin7 staining pattern showed small rings surrounding mossy fibers in wild-type CG, while it changed to the dot/spot pattern inside small rings detected with CADM1 staining in MPP2-deficient CG. These results indicate that MPP2 influences the distribution of Lin7 to synaptic cell membranes at postsynaptic regions in granule cells at CG, at which electric signals enter the cerebellum.
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Affiliation(s)
- Tomoki Yamada
- Health Science Division, Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan
| | - Yurika Saitoh
- Health Science Division, Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan
- Center for Medical Education, Teikyo University of Science, Adachi-ku, Tokyo, Japan
| | - Kiyokazu Kametani
- Health Science Division, Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan
| | - Akio Kamijo
- Health Science Division, Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan
- Division of Basic and Clinical Medicine, Nagano College of Nursing, Komagane, Nagano, Japan
| | - Takeharu Sakamoto
- Department of Cancer Biology, Institute of Biomedical Science, Kansai Medical University, Hirakata, Osaka, Japan
| | - Nobuo Terada
- Health Science Division, Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan.
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18
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Excoffon KJDA, Avila CL, Alghamri MS, Kolawole AO. The magic of MAGI-1: A scaffolding protein with multi signalosomes and functional plasticity. Biol Cell 2022; 114:185-198. [PMID: 35389514 DOI: 10.1111/boc.202200014] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/21/2022] [Revised: 04/01/2022] [Accepted: 04/04/2022] [Indexed: 11/29/2022]
Abstract
MAGI-1 is a critical cellular scaffolding protein with over 110 different cellular and microbial protein interactors. Since the discovery of MAGI-1 in 1997, MAGI-1 has been implicated in diverse cellular functions such as polarity, cell-cell communication, neurological processes, kidney function, and a host of diseases including cancer and microbial infection. Additionally, MAGI-1 has undergone nomenclature changes in response to the discovery of an additional PDZ domain, leading to lack of continuity in the literature. We address the nomenclature of MAGI-1 as well as summarize many of the critical functions of the known interactions. Given the importance of many of the interactors, such as human papillomavirus E6, the Coxsackievirus and adenovirus receptor (CAR), and PTEN, the enhancement or disruption of MAGI-based interactions has the potential to affect cellular functions that can potentially be harnessed as a therapeutic strategy for a variety of diseases.
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Affiliation(s)
| | - Christina L Avila
- Department of Biological Sciences, Wright State University, Dayton, Ohio, USA
| | - Mahmoud S Alghamri
- Department of Biological Sciences, Wright State University, Dayton, Ohio, USA
| | - Abimbola O Kolawole
- Department of Biological Sciences, Wright State University, Dayton, Ohio, USA
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19
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Matsuda M, Chu CW, Sokol SY. Lmo7 recruits myosin II heavy chain to regulate actomyosin contractility and apical domain size in Xenopus ectoderm. Development 2022; 149:275389. [PMID: 35451459 PMCID: PMC9188752 DOI: 10.1242/dev.200236] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2021] [Accepted: 03/30/2022] [Indexed: 11/20/2022]
Abstract
ABSTRACT
Apical constriction, or a reduction in size of the apical domain, underlies many morphogenetic events during development. Actomyosin complexes play an essential role in apical constriction; however, the detailed analysis of molecular mechanisms is still pending. Here, we show that Lim domain only protein 7 (Lmo7), a multidomain adaptor at apical junctions, promotes apical constriction in the Xenopus superficial ectoderm, whereas apical domain size increases in Lmo7-depleted cells. Lmo7 is primarily localized at apical junctions and promotes the formation of the dense circumferential actomyosin belt. Strikingly, Lmo7 binds non-muscle myosin II (NMII) and recruits it to apical junctions and the apical cortex. This NMII recruitment is essential for Lmo7-mediated apical constriction. Lmo7 knockdown decreases NMIIA localization at apical junctions and delays neural tube closure in Xenopus embryos. Our findings suggest that Lmo7 serves as a scaffold that regulates actomyosin contractility and apical domain size.
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Affiliation(s)
- Miho Matsuda
- Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
| | - Chih-Wen Chu
- Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
| | - Sergei Y. Sokol
- Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
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Diagbouga MR, Morel S, Cayron AF, Haemmerli J, Georges M, Hierck BP, Allémann E, Lemeille S, Bijlenga P, Kwak BR. Primary cilia control endothelial permeability by regulating expression and location of junction proteins. Cardiovasc Res 2022; 118:1583-1596. [PMID: 33974072 PMCID: PMC9074981 DOI: 10.1093/cvr/cvab165] [Citation(s) in RCA: 11] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/18/2020] [Accepted: 05/09/2021] [Indexed: 12/12/2022] Open
Abstract
AIMS Wall shear stress (WSS) determines intracranial aneurysm (IA) development. Polycystic kidney disease (PKD) patients have a high IA incidence and risk of rupture. Dysfunction/absence of primary cilia in PKD endothelial cells (ECs) may impair mechano-transduction of WSS and favour vascular disorders. The molecular links between primary cilia dysfunction and IAs are unknown. METHODS AND RESULTS Wild-type and primary cilia-deficient Tg737orpk/orpk arterial ECs were submitted to physiological (30 dynes/cm2) or aneurysmal (2 dynes/cm2) WSS, and unbiased transcriptomics were performed. Tg737orpk/orpk ECs displayed a fivefold increase in the number of WSS-responsive genes compared to wild-type cells. Moreover, we observed a lower trans-endothelial resistance and a higher endothelial permeability, which correlated with disorganized intercellular junctions in Tg737orpk/orpk cells. We identified ZO-1 as a central regulator of primary cilia-dependent endothelial junction integrity. Finally, clinical and histological characteristics of IAs from non-PKD and PKD patients were analysed. IAs in PKD patients were more frequently located in the middle cerebral artery (MCA) territory than in non-PKD patients. IA domes from the MCA of PKD patients appeared thinner with less collagen and reduced endothelial ZO-1 compared with IA domes from non-PKD patients. CONCLUSION Primary cilia dampen the endothelial response to aneurysmal low WSS. In absence of primary cilia, ZO-1 expression levels are reduced, which disorganizes intercellular junctions resulting in increased endothelial permeability. This altered endothelial function may not only contribute to the severity of IA disease observed in PKD patients, but may also serve as a potential diagnostic tool to determine the vulnerability of IAs.
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Affiliation(s)
- Mannekomba R Diagbouga
- Department of Pathology and Immunology, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
| | - Sandrine Morel
- Department of Pathology and Immunology, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
- Neurosurgery Division, Department of Clinical Neurosciences, Geneva University Hospitals and University of Geneva, Rue Gabrielle-Perret-Gentil 4, CH-1211 Geneva, Switzerland
| | - Anne F Cayron
- Department of Pathology and Immunology, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
- School of Pharmaceutical Sciences, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
- Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
| | - Julien Haemmerli
- Neurosurgery Division, Department of Clinical Neurosciences, Geneva University Hospitals and University of Geneva, Rue Gabrielle-Perret-Gentil 4, CH-1211 Geneva, Switzerland
| | - Marc Georges
- Neurosurgery Division, Department of Clinical Neurosciences, Geneva University Hospitals and University of Geneva, Rue Gabrielle-Perret-Gentil 4, CH-1211 Geneva, Switzerland
| | - Beerend P Hierck
- Department of Anatomy and Embryology, Leiden University Medical Center, Eindhovenweg 20, 2333ZC Leiden, the Netherlands
| | - Eric Allémann
- School of Pharmaceutical Sciences, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
- Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
| | - Sylvain Lemeille
- Department of Pathology and Immunology, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
| | - Philippe Bijlenga
- Neurosurgery Division, Department of Clinical Neurosciences, Geneva University Hospitals and University of Geneva, Rue Gabrielle-Perret-Gentil 4, CH-1211 Geneva, Switzerland
| | - Brenda R Kwak
- Department of Pathology and Immunology, University of Geneva, Rue Michel-Servet 1, CH-1211 Geneva, Switzerland
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21
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Vaid S, Huttner WB. Progenitor-Based Cell Biological Aspects of Neocortex Development and Evolution. Front Cell Dev Biol 2022; 10:892922. [PMID: 35602606 PMCID: PMC9119302 DOI: 10.3389/fcell.2022.892922] [Citation(s) in RCA: 9] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/09/2022] [Accepted: 04/14/2022] [Indexed: 11/13/2022] Open
Abstract
During development, the decision of stem and progenitor cells to switch from proliferation to differentiation is of critical importance for the overall size of an organ. Too early a switch will deplete the stem/progenitor cell pool, and too late a switch will not generate the required differentiated cell types. With a focus on the developing neocortex, a six-layered structure constituting the major part of the cerebral cortex in mammals, we discuss here the cell biological features that are crucial to ensure the appropriate proliferation vs. differentiation decision in the neural progenitor cells. In the last two decades, the neural progenitor cells giving rise to the diverse types of neurons that function in the neocortex have been intensely investigated for their role in cortical expansion and gyrification. In this review, we will first describe these different progenitor types and their diversity. We will then review the various cell biological features associated with the cell fate decisions of these progenitor cells, with emphasis on the role of the radial processes emanating from these progenitor cells. We will also discuss the species-specific differences in these cell biological features that have allowed for the evolutionary expansion of the neocortex in humans. Finally, we will discuss the emerging role of cell cycle parameters in neocortical expansion.
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Affiliation(s)
- Samir Vaid
- Department of Basic Neurosciences, University of Geneva, Geneva, Switzerland
- *Correspondence: Samir Vaid, ; Wieland B. Huttner,
| | - Wieland B. Huttner
- Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
- *Correspondence: Samir Vaid, ; Wieland B. Huttner,
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22
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Beckmann A, Recktenwald J, Ferdinand A, Grißmer A, Meier C. First Responders to Hyperosmotic Stress in Murine Astrocytes: Connexin 43 Gap Junctions Are Subject to an Immediate Ultrastructural Reorganization. BIOLOGY 2021; 10:biology10121307. [PMID: 34943223 PMCID: PMC8698406 DOI: 10.3390/biology10121307] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/20/2021] [Revised: 12/04/2021] [Accepted: 12/06/2021] [Indexed: 11/16/2022]
Abstract
Simple Summary Gap junctions are intercellular channels that provide the means for direct transport of small molecules, ions, and water between connected cells. With these functions, gap junctions are essential for the maintenance of astrocytic homeostasis and of particular importance in the context of pathophysiological disbalances. These include the hyperosmolar hyperglycemic syndrome or the pathology after brain trauma. We demonstrate that short-term hyperosmolarity reduces intercellular communication via gap junctions. These functional changes coincide with the transformation of gap junction ultrastructure as evidenced by freeze-fracture replica immunolabeling and transmission electron microscopy. The hyperosmolarity-induced immediate changes in the ultrastructural assembly of connexons, the protein constituents of gap junction channels, have not been described in astrocytes before and are revealing the coherence of structure and function in gap junctions. Phosphorylation of Connexin 43, the main gap junction protein in astrocytes, at amino acid 368 (Serine) might link the two. Abstract In a short-term model of hyperosmotic stress, primary murine astrocytes were stimulated with a hyperosmolar sucrose solution for five minutes. Astrocytic gap junctions, which are mainly composed of Connexin (Cx) 43, displayed immediate ultrastructural changes, demonstrated by freeze–fracture replica immunogold labeling: their area, perimeter, and distance of intramembrane particles increased, whereas particle numbers per area decreased. Ultrastructural changes were, however, not accompanied by changes in Cx43 mRNA expression. In contrast, transcription of the gap junction regulator zonula occludens (ZO) protein 1 significantly increased, whereas its protein expression was unaffected. Phosphorylation of Serine (S) 368 of the Cx43 C–terminus has previously been associated with gap junction disassembly and reduction in gap junction communication. Hyperosmolar sucrose treatment led to enhanced phosphorylation of Cx43S368 and was accompanied by inhibition of gap junctional intercellular communication, demonstrated by a scrape loading-dye transfer assay. Taken together, Cx43 gap junctions are fast reacting elements in response to hyperosmolar challenges and can therefore be considered as one of the first responders to hyperosmolarity. In this process, phosphorylation of Cx43S368 was associated with disassembly of gap junctions and inhibition of their function. Thus, modulation of the gap junction assembly might represent a target in the treatment of brain edema or trauma.
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23
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Sugawara T, Furuse K, Otani T, Wakayama T, Furuse M. Angulin-1 seals tricellular contacts independently of tricellulin and claudins. J Cell Biol 2021; 220:e202005062. [PMID: 34269802 PMCID: PMC8289698 DOI: 10.1083/jcb.202005062] [Citation(s) in RCA: 31] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2020] [Revised: 04/24/2021] [Accepted: 06/21/2021] [Indexed: 12/24/2022] Open
Abstract
Tricellular tight junctions (tTJs) are specialized tight junctions (TJs) that seal the intercellular space at tricellular contacts (TCs), where the vertices of three epithelial cells meet. Tricellulin and angulin family membrane proteins are known constituents of tTJs, but the molecular mechanism of tTJ formation remains elusive. Here, we investigated the roles of angulin-1 and tricellulin in tTJ formation in MDCK II cells by genome editing. Angulin-1-deficient cells lost the plasma membrane contact at TCs with impaired epithelial barrier function. The C terminus of angulin-1 bound to the TJ scaffold protein ZO-1, and disruption of their interaction influenced the localization of claudins at TCs, but not the tricellular sealing. Strikingly, the plasma membrane contact at TCs was formed in tricellulin- or claudin-deficient cells. These findings demonstrate that angulin-1 is responsible for the plasma membrane seal at TCs independently of tricellulin and claudins.
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Affiliation(s)
- Taichi Sugawara
- Division of Cell Structure, National Institute for Physiological Sciences, National Institute of Natural Sciences, Okazaki, Aichi, Japan
- Department of Physiological Sciences, The Graduate University for Advanced Studies, SOKENDAI, Okazaki, Aichi, Japan
- Department of Histology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan
| | - Kyoko Furuse
- Division of Cell Structure, National Institute for Physiological Sciences, National Institute of Natural Sciences, Okazaki, Aichi, Japan
| | - Tetsuhisa Otani
- Division of Cell Structure, National Institute for Physiological Sciences, National Institute of Natural Sciences, Okazaki, Aichi, Japan
- Department of Physiological Sciences, The Graduate University for Advanced Studies, SOKENDAI, Okazaki, Aichi, Japan
| | - Tomohiko Wakayama
- Department of Histology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan
| | - Mikio Furuse
- Division of Cell Structure, National Institute for Physiological Sciences, National Institute of Natural Sciences, Okazaki, Aichi, Japan
- Department of Physiological Sciences, The Graduate University for Advanced Studies, SOKENDAI, Okazaki, Aichi, Japan
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24
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Assimakopoulos SF, Mastronikolis S, DE Lastic AL, Aretha D, Papageorgiou D, Chalkidi T, Oikonomou I, Triantos C, Mouzaki A, Marangos M. Intestinal Barrier Biomarker ZO1 and Endotoxin Are Increased in Blood of Patients With COVID-19-associated Pneumonia. In Vivo 2021; 35:2483-2488. [PMID: 34182534 DOI: 10.21873/invivo.12528] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/06/2021] [Revised: 05/21/2021] [Accepted: 05/25/2021] [Indexed: 02/07/2023]
Abstract
BACKGROUND/AIM The present study was undertaken to investigate (i) whether hospitalized patients with COVID-19 pneumonia present intestinal barrier dysfunction with consequent translocation of endotoxin into the systemic circulation and (ii) whether intestinal barrier biomarkers have any prognostic role in terms of progression to severe respiratory failure. PATIENTS AND METHODS In this prospective study, 22 patients with COVID-19-associated pneumonia and 19 patients with non-COVID-19-related community-acquired pneumonia (CAP group) were studied while 12 healthy persons comprised the control group. Blood samples were collected on admission and analysed for serum levels of endotoxin and zonula occludens-1 (ZO1). Clinical courses regarding progression to severe respiratory failure (SRF) requiring mechanical ventilation were recorded. RESULTS Patients with COVID-19-associated pneumonia and patients with CAP presented significantly higher serum endotoxin and ZO1 concentrations on admission as compared to healthy controls. There was no difference in endotoxin levels between patients with COVID-19-related pneumonia and patients with CAP. In patients with COVID-19-related pneumonia, serum endotoxin concentrations were positively correlated with C-reactive protein and ferritin values. There were no significant differences in serum endotoxin and ZO1 concentrations between patients with severe and not severe COVID-19-related pneumonia, nor between patients who developed SRF and those who did not Conclusion: Patients with COVID-19-related pneumonia present intestinal barrier dysfunction leading to systemic endotoxemia. Admission values of endotoxin and ZO1 do not have any prognostic role for progression to SRF.
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Affiliation(s)
| | | | - Anne-Lise DE Lastic
- Division of Hematology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Diamanto Aretha
- Department of Anesthesiology and Intensive Care Medicine, University of Patras Medical School, Patras, Greece
| | - Dimitris Papageorgiou
- Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Theodora Chalkidi
- Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Ioanna Oikonomou
- Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Christos Triantos
- Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Athanasia Mouzaki
- Division of Hematology, Department of Internal Medicine, University of Patras Medical School, Patras, Greece
| | - Markos Marangos
- Department of Internal Medicine, University of Patras Medical School, Patras, Greece
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25
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Takada T, Takikawa H, Sawada N, Higuchi R, Nagamachi Y, Isaji S, Yoshida M, Yamamoto M. Cholangio-venous reflux of biliary contents through paracellular pathways between hepatocytes in patients with acute cholangitis. JOURNAL OF HEPATO-BILIARY-PANCREATIC SCIENCES 2021; 28:508-514. [PMID: 33720522 DOI: 10.1002/jhbp.937] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/19/2020] [Revised: 02/14/2021] [Accepted: 03/05/2021] [Indexed: 11/07/2022]
Abstract
BACKGROUND We re-analyzed data on cholangio-venous reflux from a clinical study conducted prospectively on 22 patients in 1974. METHOD Direct cholangiography was performed with indocyanine green (ICG) mixed into UrographinR under monitoring of intrabiliary pressure, and the participants were allocated to three groups according to whether ICG leakage into the blood, signs of infection, or both, were present. RESULTS The intrabiliary pressure of six patients negative for both ICG leakage and signs of infection was approximately 19.5 (median, [range 18-22]) cmH2 O. In contrast, for the five patients positive for ICG leakage but negative for signs of infection, the intrabiliary pressure was higher (median 32.0 [range 27-41) cmH2 O]. The 11 patients positive for both ICG leakage and signs of infection had the highest intrabiliary pressure (median 48.0 [range 33-77] cmH2 O). Our analyses revealed that, as the intrabiliary pressure increased, the status of ICG leakage and signs of infection appeared in a stepwise fashion. CONCLUSION Our findings suggest that the tight junctions sealing the bile canaliculi deteriorated with increasing intrabiliary pressure, resulting in reflux of the biliary contents into the vascular system via paracellular pathways between hepatocytes.
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Affiliation(s)
- Tadahiro Takada
- Department of Surgery, Teikyo University School of Medicine, Tokyo, Japan
| | - Hajime Takikawa
- Department of Medicine, Teikyo University School of Medicine, Tokyo, Japan
| | - Norimasa Sawada
- Second Department of Pathology, Sapporo Medical University, Sapporo, Japan
| | - Ryota Higuchi
- Department of Surgery, Institute of Gastroenterology, Tokyo Women's Medical University, Tokyo, Japan
| | - Yukiko Nagamachi
- Department of Medicine, Teikyo University School of Medicine, Tokyo, Japan
| | - Shuji Isaji
- Department of Hepatobiliary Pancreatic Surgery and Transplantation, Mie University, Mie, Japan
| | - Masahiro Yoshida
- Department of Hepatobiliary Pancreatic and Gastrointestinal Surgery, School of Medicine, International University of Health and Welfare, Ichikawa, Japan
| | - Masakazu Yamamoto
- Department of Surgery, Institute of Gastroenterology, Tokyo Women's Medical University, Tokyo, Japan
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26
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Rouaud F, Sluysmans S, Flinois A, Shah J, Vasileva E, Citi S. Scaffolding proteins of vertebrate apical junctions: structure, functions and biophysics. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES 2020; 1862:183399. [DOI: 10.1016/j.bbamem.2020.183399] [Citation(s) in RCA: 24] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/13/2020] [Revised: 06/05/2020] [Accepted: 06/11/2020] [Indexed: 12/11/2022]
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Barathi S, Aruljothi KN, Karthik C, Padikasan IA. Optimization for enhanced ecofriendly decolorization and detoxification of Reactive Blue160 textile dye by Bacillus subtilis. ACTA ACUST UNITED AC 2020; 28:e00522. [PMID: 32963974 PMCID: PMC7490544 DOI: 10.1016/j.btre.2020.e00522] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2019] [Revised: 06/28/2020] [Accepted: 08/28/2020] [Indexed: 12/22/2022]
Abstract
The bacterial strain capable of decolorization and detoxification of the Reactive Blue 160 dye was isolated from a dye waste disposal site of Tirupur textile industries. The bacterial strain was screened and selected based on its decolorization capability of RB 160dye, which was identified as Bacillus subtilis by 16S rRNA sequencing. The strain was tested for the decolorization potential under different physio-chemical experimental conditions (pH, temperature, agitation, non-agitation) and observed a complete decolorization at pH 7 and 35 °C under shaking condition within 48 h of time. The enzymes such as, Lignin peroxidase, azoreductase and NADH-DCI were significantly induced in the strain during the decolorization of RB160 dye. Phytotoxicity and microbial toxicity studies revealed that the decolorized product of RB160 dye is less toxic to the plants and microbes. Thus, our results recommend the prospective use of B subtilis in bioremediation of RB160 dye.
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Affiliation(s)
- Selvaraj Barathi
- School of Environmental Science and Engineering, Sun Yat-Sen University, Guangzhou, People's Republic of China.,Plant and Microbial Biotechnology Laboratory, Department of Biotechnology, School of Biosciences, Periyar University, Salem, 636 011, Tamil Nadu, India
| | - K N Aruljothi
- Department of GeneticEngineering, SRM Institute of Science and Technology, India
| | - Chinnannan Karthik
- College of Agriculture and Biotechnology, Institute of Crop Science, Zhejiang University, China
| | - Indra Arulselvi Padikasan
- Plant and Microbial Biotechnology Laboratory, Department of Biotechnology, School of Biosciences, Periyar University, Salem, 636 011, Tamil Nadu, India
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28
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Liu F, Xu T, Peng S, Adelman RA, Rizzolo LJ. Claudins regulate gene and protein expression of the retinal pigment epithelium independent of their association with tight junctions. Exp Eye Res 2020; 198:108157. [PMID: 32712183 DOI: 10.1016/j.exer.2020.108157] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/20/2020] [Revised: 06/18/2020] [Accepted: 07/20/2020] [Indexed: 01/14/2023]
Abstract
Claudin-19 is the major claudin in the tight junctions of the retinal pigment epithelium (RPE). Claudin-3 is also uniformly expressed albeit in lesser amounts. Besides modulating transepithelial diffusion, claudins modulate gene expression. The absence of claudin-19 and claudin-3 in the RPE cell lines, ARPE-19 and hTERT-RPE-1, provide an opportunity to examine whether exogenous claudins regulate gene expression in the absence of tight junctions. Quantitative RT-PCR was used to compare gene expression in ARPE-19 and hTERT-RPE-1 with that of highly differentiated, human fetal RPE. Claudin-19 and claudin-3 were exogenously expressed using an adenoviral vector. The transepithelial electrical resistance (TER) was measured using Endohm electrodes, and the effects of claudin on the actin cytoskeleton were determined by immunocytochemistry. The effect of claudin on gene expression was examined by quantitative RT-PCR and western blotting. Aside from claudin-19 and claudin-3, ARPE-19 and hTERT-RPE-1 expressed most junction-associated mRNAs in amounts comparable to human fetal RPE, but some RPE signature and maturation genes were under-expressed. Unlike ARPE-19, hTERT-RPE-1 failed to form tight junctions or develop a TER. Claudins exogenously expressed in hTERT-RPE-1 failed to crystalize an apical junctional complex. Actin filaments were not redistributed from stress fibers to cortical bands, and a TER was not established. In hTERT-RPE-1, claudins were found only in internal vesicular-like structures. Nonetheless, claudins increased the expression of the mRNAs for a collection of RPE-enriched proteins. Claudin-19 and claudin-3 had different effects on gene and protein expression indicating activation of overlapping, but distinct, signaling pathways. A major difference was the ability of claudin-19 to affect steady-state levels of ADAM9 and tyrosinase in ARPE-19. In conclusion, claudins can increase the barrier function of a pre-existing apical junctional complex, but on its own it cannot recruit tight junction proteins to form a complex de novo. Many effects of claudin on gene expression did not require an association with the apical junctional complex. Although claudin-19 shared many effects with claudin-3, claudin-19 exerted unique effects on the maturation of RPE.
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Affiliation(s)
- Fanfei Liu
- Aier School of Ophthalmology, Central South University, Changsha, China; Department of Surgery, Yale University, New Haven, USA; Department of Ophthalmology and Visual Science, Yale University, New Haven, USA
| | - Tao Xu
- Aier School of Ophthalmology, Central South University, Changsha, China; Department of Surgery, Yale University, New Haven, USA; Department of Ophthalmology and Visual Science, Yale University, New Haven, USA
| | - Shaomin Peng
- Aier School of Ophthalmology, Central South University, Changsha, China.
| | - Ron A Adelman
- Department of Ophthalmology and Visual Science, Yale University, New Haven, USA
| | - Lawrence J Rizzolo
- Department of Surgery, Yale University, New Haven, USA; Department of Ophthalmology and Visual Science, Yale University, New Haven, USA.
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Fan J, Li TJ, Zhao XH. Barrier-promoting efficiency of two bioactive flavonols quercetin and myricetin on rat intestinal epithelial (IEC-6) cells via suppressing Rho activation. RSC Adv 2020; 10:27249-27258. [PMID: 35516969 PMCID: PMC9055572 DOI: 10.1039/d0ra04162a] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/09/2020] [Accepted: 07/15/2020] [Indexed: 01/13/2023] Open
Abstract
Polyphenols are beneficial to human health because of their bio-activities. In this study, two flavonols quercetin and myricetin with or without heat treatment at 100 °C for 30 min were assessed for their barrier-promoting efficiency in rat intestinal epithelial (IEC-6) cells. The results indicated that the heated and unheated flavonols at dose levels of 2.5-20 μmol L-1 had a nontoxic effect on the cells treated for 24 and 48 h but enhanced the values of cell viability larger than 100% (especially at a dose level of 5 μmol L-1). Moreover, the cells exposed to these flavonols of 5 μmol L-1 for 24 and 48 h had improved barrier integrity compared to the control cells without any flavonol treatment, reflected by enhanced transepithelial electrical resistance and anti-bacterial effect but decreased paracellular permeability and bacterial translocation. Moreover, the results from both mRNA and protein expression verified 1.1-3.4 fold up-regulation of zonula occludens-1, occludin, and claudin-1 that are critical to tight junctions and barrier function of cells. Furthermore, the expression of other two proteins RhoA and ROCK in the treated cells was also down-regulated, demonstrating suppressed Rho activation and consequently barrier promotion via the RhoA/ROCK signaling pathway. Overall quercetin, due to its lower molecular polarity, mostly gave higher barrier-promoting efficiency than myricetin, while the heated flavonols were always less efficient than the unheated counterparts to promote barrier integrity of IEC-6 cells. It is thus highlighted that flavonols can provide barrier-promoting effects on intestinal epithelial cells with a promoting efficiency dependent on flavonol polarity; however, heat treatment especially excessive heat treatment of plant foods might lead to damaged flavonol activity.
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Affiliation(s)
- Jing Fan
- Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University 150030 Harbin PR China
| | - Tie-Jing Li
- College of Light Industry, Liaoning University 110136 Shenyang PR China
| | - Xin-Huai Zhao
- Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University 150030 Harbin PR China
- School of Biology and Food Engineering, Guangdong University of Petrochemical Technology 525000 Maoming PR China
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30
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Zhang J, Vincent KP, Peter AK, Klos M, Cheng H, Huang SM, Towne JK, Ferng D, Gu Y, Dalton ND, Chan Y, Li R, Peterson KL, Chen J, McCulloch AD, Knowlton KU, Ross RS. Cardiomyocyte Expression of ZO-1 Is Essential for Normal Atrioventricular Conduction but Does Not Alter Ventricular Function. Circ Res 2020; 127:284-297. [PMID: 32345129 DOI: 10.1161/circresaha.119.315539] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
RATIONALE ZO-1 (Zonula occludens-1), a plasma membrane-associated scaffolding protein regulates signal transduction, transcription, and cellular communication. Global deletion of ZO-1 in the mouse is lethal by embryonic day 11.5. The function of ZO-1 in cardiac myocytes (CM) is largely unknown. OBJECTIVE To determine the function of CM ZO-1 in the intact heart, given its binding to other CM proteins that have been shown instrumental in normal cardiac conduction and function. METHODS AND RESULTS We generated ZO-1 CM-specific knockout (KO) mice using α-Myosin Heavy Chain-nuclear Cre (ZO-1cKO) and investigated physiological and electrophysiological function by echocardiography, surface ECG and conscious telemetry, intracardiac electrograms and pacing, and optical mapping studies. ZO-1cKO mice were viable, had normal Mendelian ratios, and had a normal lifespan. Ventricular morphometry and function were not significantly different between the ZO-1cKO versus control (CTL) mice, basally in young or aged mice, or even when hearts were subjected to hemodynamic loading. Atrial mass was increased in ZO-1cKO. Electrophysiological and optical mapping studies indicated high-grade atrioventricular (A-V) block in ZO-1cKO comparing to CTL hearts. While ZO-1-associated proteins such as vinculin, connexin 43, N-cadherin, and α-catenin showed no significant change with the loss of ZO-1, Connexin-45 and Coxsackie-adenovirus (CAR) proteins were reduced in atria of ZO-1cKO. Further, with loss of ZO-1, ZO-2 protein was increased significantly in ventricular CM in a presumed compensatory manner but was still not detected in the AV nodal myocytes. Importantly, the expression of the sodium channel protein NaV1.5 was altered in AV nodal cells of the ZO-1cKO versus CTL. CONCLUSIONS ZO-1 protein has a unique physiological role in cardiac nodal tissue. This is in alignment with its known interaction with CAR and Cx45, and a new function in regulating the expression of NaV1.5 in AV node. Uniquely, ZO-1 is dispensable for function of the working myocardium.
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Affiliation(s)
- Jianlin Zhang
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Kevin P Vincent
- Department of Bioengineering (K.P.V., A.D.M.), University of California San Diego, La Jolla, CA
| | - Angela K Peter
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Matthew Klos
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Hongqiang Cheng
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Selina M Huang
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Jordan K Towne
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Debbie Ferng
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Yusu Gu
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Nancy D Dalton
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Yunghang Chan
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Ruixia Li
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Kirk L Peterson
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Ju Chen
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
| | - Andrew D McCulloch
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
- Department of Bioengineering (K.P.V., A.D.M.), University of California San Diego, La Jolla, CA
| | | | - Robert S Ross
- From the Department of Medicine (J.Z., A.K.P., M.K., H.C., S.M.H., J.K.T., D.F., Y.G., N.D.D., Y.C., J.K.T., D.F., Y.G., N.D.D., Y.C., R.L., K.L.P., J.C., A.D.M., R.S.R.), University of California San Diego, La Jolla, CA
- Veterans Administration Healthcare, Cardiology Section, San Diego, CA (R.S.R.)
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Shi Y, Li R, Yang J, Li X. No tight junctions in tight junction protein-1 expressing HeLa and fibroblast cells. INTERNATIONAL JOURNAL OF PHYSIOLOGY, PATHOPHYSIOLOGY AND PHARMACOLOGY 2020; 12:70-78. [PMID: 32419902 PMCID: PMC7218737] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Subscribe] [Scholar Register] [Received: 01/06/2020] [Accepted: 04/08/2020] [Indexed: 06/11/2023]
Abstract
Tight junctions are important structures that form the barrier of cells and tissues, and they play key roles in maintaining homeostasis of our body. The backbone of the tight junction proteins are claudins, which composed more than twenty members. The tight junction protein 1 (TJP1), also called ZO-1 (Zonula Occludens-1), is one of the tight junction related proteins, and it is widely used in literature to label tight junctions. Here we showed that TJP1 (ZO-1) is highly expressed in cancerous HeLa cells, fibroblast cells, HUVEC as well as MDCK cells, while claudin-1 is highly expressed in HUVEC and MDCK cells, but not expressed in HeLa and fibroblast cells. We aimed to investigate whether tight junction is present in HeLa and fibroblast cells. We used transepithelial/transendothelial electrical resistance (TEER) to measure tight junction dynamics in these cells. The results showed that there is no TEERs in HeLa and fibroblast cells, while there is relatively high TEER in HUVEC and MDCK cells. Importantly, the TEER in MDCK cells is dramatically reduced after knockdown of TJP1 (ZO-1). These results suggest that TJP1 (ZO-1) cannot be used as a marker of tight junctions in a variety of cells, while TJP1 (ZO-1) may play an important role in regulation of tight junctions in MDCK cells.
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Affiliation(s)
- Yumeng Shi
- Eye Institute, Eye and Ear, Nose, and Throat Hospital of Fudan University83 Fenyang Road, Shanghai 200031, People’s Republic of China
| | - Rongqiang Li
- Department of Urology, Weihaiwei HospitalWeihai, Shandong, China
| | - Jin Yang
- Eye Institute, Eye and Ear, Nose, and Throat Hospital of Fudan University83 Fenyang Road, Shanghai 200031, People’s Republic of China
| | - Xinbo Li
- Ophthalmology, Casey Eye Institute, Oregon Health & Science UniversityPortland, Oregon, USA
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Li X, Acott TS, Nagy JI, Kelley MJ. ZO-1 associates with α3 integrin and connexin43 in trabecular meshwork and Schlemm's canal cells. INTERNATIONAL JOURNAL OF PHYSIOLOGY, PATHOPHYSIOLOGY AND PHARMACOLOGY 2020; 12:1-10. [PMID: 32211117 PMCID: PMC7076326] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Subscribe] [Scholar Register] [Received: 12/11/2019] [Accepted: 01/14/2020] [Indexed: 06/10/2023]
Abstract
Cellular structures that perform essential homeostatic functions include tight junctions, gap junctions, desmosomes and adherens junctions. The aqueous humor, produced by the ciliary body, passes into the anterior chamber of the eye and is filtered by the trabecular meshwork (TM), a tiny tissue found in the angle of the eye. This tissue, along with Schlemm's canal (SC) inner wall cells, is thought to control intraocular pressure (IOP) homeostasis for normal, optimal vision. The actin cytoskeleton of the tissue plays a regulatory role in maintaining IOP. One of the key risk factors for primary open angle glaucoma is persistent elevation of IOP, which compromises the optic nerve. The ZO-1 (Zonula Occludens-1), extracellular matrix protein integrins, and gap junction protein connexin43 (Cx43) are widely expressed in many different cell populations. Here, we investigated the localization and interactions of ZO-1, α3 integrin, β1 integrin, and Cx43 in cultured porcine TM and SC cells using RT-PCR, western immunoblotting and immunofluorescence labeling with confocal microscopy, along with co-immunoprecipitation. ZO-1 partially co-localized with α3 integrin, but not with β1 integrin, and co-immunoprecipitated with Cx43, as well as with α3 integrin. The association of ZO-1 with α3 integrin and Cx43 suggests that these proteins may form a multiple protein complex in porcine TM and SC cells. Since integrins interact with the actin cytoskeleton via scaffolding proteins, these results implicate junctional and scaffolding protein ZO-1 as a potential control point in regulation of IOP to normal levels for glaucoma therapy.
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Affiliation(s)
- Xinbo Li
- Department of Ophthalmology, Casey Eye Institute, Oregon Health and Science UniversityPortland, Oregon, USA
| | - Ted S Acott
- Department of Ophthalmology, Casey Eye Institute, Oregon Health and Science UniversityPortland, Oregon, USA
- Department of Chemical Physiology and Biochemistry, Oregon Health and Science UniversityPortland, Oregon, USA
| | - James I Nagy
- Department of Physiology and Pathophysiology, University of ManitobaWinnipeg, MB, Canada
| | - Mary J Kelley
- Department of Ophthalmology, Casey Eye Institute, Oregon Health and Science UniversityPortland, Oregon, USA
- Department of Integrative Bioscience, Oregon Health and Science UniversityPortland, Oregon, USA
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The Prognostic Value of Endotoxemia and Intestinal Barrier Biomarker ZO-1 in Bacteremic Sepsis. Am J Med Sci 2020; 359:100-107. [DOI: 10.1016/j.amjms.2019.10.006] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/28/2019] [Revised: 10/13/2019] [Accepted: 10/16/2019] [Indexed: 02/07/2023]
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Fasano A. All disease begins in the (leaky) gut: role of zonulin-mediated gut permeability in the pathogenesis of some chronic inflammatory diseases. F1000Res 2020; 9. [PMID: 32051759 PMCID: PMC6996528 DOI: 10.12688/f1000research.20510.1] [Citation(s) in RCA: 256] [Impact Index Per Article: 51.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Accepted: 01/22/2020] [Indexed: 12/19/2022] Open
Abstract
Improved hygiene leading to reduced exposure to microorganisms has been implicated as one possible cause for the recent "epidemic" of chronic inflammatory diseases (CIDs) in industrialized countries. That is the essence of the hygiene hypothesis that argues that rising incidence of CIDs may be, at least in part, the result of lifestyle and environmental changes that have made us too "clean" for our own good, so causing changes in our microbiota. Apart from genetic makeup and exposure to environmental triggers, inappropriate increase in intestinal permeability (which may be influenced by the composition of the gut microbiota), a "hyper-belligerent" immune system responsible for the tolerance-immune response balance, and the composition of gut microbiome and its epigenetic influence on the host genomic expression have been identified as three additional elements in causing CIDs. During the past decade, a growing number of publications have focused on human genetics, the gut microbiome, and proteomics, suggesting that loss of mucosal barrier function, particularly in the gastrointestinal tract, may substantially affect antigen trafficking, ultimately influencing the close bidirectional interaction between gut microbiome and our immune system. This cross-talk is highly influential in shaping the host gut immune system function and ultimately shifting genetic predisposition to clinical outcome. This observation led to a re-visitation of the possible causes of CIDs epidemics, suggesting a key pathogenic role of gut permeability. Pre-clinical and clinical studies have shown that the zonulin family, a group of proteins modulating gut permeability, is implicated in a variety of CIDs, including autoimmune, infective, metabolic, and tumoral diseases. These data offer novel therapeutic targets for a variety of CIDs in which the zonulin pathway is implicated in their pathogenesis.
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Affiliation(s)
- Alessio Fasano
- Mucosal Immunology and Biology Research Center, Center for Celiac Research and Treatment and Division of Pediatric Gastroenterology and Nutrition, Massachusetts General Hospital for Children, Boston, Massachusetts, USA.,European Biomedical Research Institute of Salerno, Salerno, Italy
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35
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Tanaka F, Uda M, Hirose Y, Hirai Y. Restoration of calcium-induced differentiation potential and tight junction formation in HaCaT keratinocytes by functional attenuation of overexpressed high mobility group box-1 protein. Cytotechnology 2020; 72:165-174. [PMID: 31916113 DOI: 10.1007/s10616-019-00367-6] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2019] [Accepted: 12/30/2019] [Indexed: 12/15/2022] Open
Abstract
HaCaT cells have been widely used as undifferentiated epidermal keratinocytes, since these non-tumorigenic cells can be readily maintained in conventional medium and partly retain epidermal differentiation potential upon stimulation with high concentration of calcium. In contrast to primary epidermal keratinocytes, however, these cells never form tight junction (TJ), a specific structure in highly differentiated keratinocytes, solely by the differentiation stimulation. Here, we show that HaCaT cells secrete a considerable amount of high mobility group box-1 protein (HMGB1), one of major inflammatory mediator, which appeared to be responsible, at least in part, for such aberrant differentiation response. So far, inhibition of c-Jun N-terminal kinase (JNK) in high calcium medium has been supposed to be the only way to induce TJ formations in HaCaT cells; however, SP600125, a potent inhibitor of JNK showed cytostatic effects and clearly attenuated epidermal differentiation and stratification. In contrast, dipotassium glycyrrhizate (GK2), a soluble analogue of HMGB1-blocker Glycyrrhizin, down-regulated interferon-β, a typical inflammatory cytokine induced by secreted HMGB1, and accelerated differentiation responses to the calcium treatment in these cells. In addition, GK2-treatmenrt resulted in the formation of double cell layers in cultured HaCaT cells, where the stratified upper cells transiently accumulated TJ proteins at the cell-cell contact sites. These results highlight the importance of attenuation of secreted HMGB1-signals in cultured HaCaT cells for studies of functional keratinocytes.
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Affiliation(s)
- Fumika Tanaka
- Department of Biomedical Chemistry, Kwansei Gakuin University, 2-1 Gakuen, Sanda, 669-1337, Japan
| | - Minori Uda
- Department of Biomedical Chemistry, Kwansei Gakuin University, 2-1 Gakuen, Sanda, 669-1337, Japan
| | - Yuina Hirose
- Department of Biomedical Chemistry, Kwansei Gakuin University, 2-1 Gakuen, Sanda, 669-1337, Japan
| | - Yohei Hirai
- Department of Biomedical Chemistry, Kwansei Gakuin University, 2-1 Gakuen, Sanda, 669-1337, Japan.
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Fields MA, Del Priore LV, Adelman RA, Rizzolo LJ. Interactions of the choroid, Bruch's membrane, retinal pigment epithelium, and neurosensory retina collaborate to form the outer blood-retinal-barrier. Prog Retin Eye Res 2019; 76:100803. [PMID: 31704339 DOI: 10.1016/j.preteyeres.2019.100803] [Citation(s) in RCA: 88] [Impact Index Per Article: 14.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/02/2019] [Revised: 10/26/2019] [Accepted: 10/28/2019] [Indexed: 01/10/2023]
Abstract
The three interacting components of the outer blood-retinal barrier are the retinal pigment epithelium (RPE), choriocapillaris, and Bruch's membrane, the extracellular matrix that lies between them. Although previously reviewed independently, this review integrates these components into a more wholistic view of the barrier and discusses reconstitution models to explore the interactions among them. After updating our understanding of each component's contribution to barrier function, we discuss recent efforts to examine how the components interact. Recent studies demonstrate that claudin-19 regulates multiple aspects of RPE's barrier function and identifies a barrier function whereby mutations of claudin-19 affect retinal development. Co-culture approaches to reconstitute components of the outer blood-retinal barrier are beginning to reveal two-way interactions between the RPE and choriocapillaris. These interactions affect barrier function and the composition of the intervening Bruch's membrane. Normal or disease models of Bruch's membrane, reconstituted with healthy or diseased RPE, demonstrate adverse effects of diseased matrix on RPE metabolism. A stumbling block for reconstitution studies is the substrates typically used to culture cells are inadequate substitutes for Bruch's membrane. Together with human stem cells, the alternative substrates that have been designed offer an opportunity to engineer second-generation culture models of the outer blood-retinal barrier.
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Affiliation(s)
- Mark A Fields
- Department of Ophthalmology and Visual Science, Yale University School of Medicine, PO Box 208061, New Haven, CT, 06520-8061, USA
| | - Lucian V Del Priore
- Department of Ophthalmology and Visual Science, Yale University School of Medicine, PO Box 208061, New Haven, CT, 06520-8061, USA
| | - Ron A Adelman
- Department of Ophthalmology and Visual Science, Yale University School of Medicine, PO Box 208061, New Haven, CT, 06520-8061, USA
| | - Lawrence J Rizzolo
- Department of Ophthalmology and Visual Science, Yale University School of Medicine, PO Box 208061, New Haven, CT, 06520-8061, USA; Department of Surgery, Yale University School of Medicine, PO Box 208062, New Haven, CT, 06520-8062, USA.
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Otani T, Nguyen TP, Tokuda S, Sugihara K, Sugawara T, Furuse K, Miura T, Ebnet K, Furuse M. Claudins and JAM-A coordinately regulate tight junction formation and epithelial polarity. J Cell Biol 2019; 218:3372-3396. [PMID: 31467165 PMCID: PMC6781433 DOI: 10.1083/jcb.201812157] [Citation(s) in RCA: 145] [Impact Index Per Article: 24.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/26/2018] [Revised: 06/14/2019] [Accepted: 07/24/2019] [Indexed: 01/03/2023] Open
Abstract
Tight junctions (TJs) establish the epithelial barrier and are thought to form a membrane fence to regulate epithelial polarity, although the roles of TJs in epithelial polarity remain controversial. Claudins constitute TJ strands in conjunction with the cytoplasmic scaffolds ZO-1 and ZO-2 and play pivotal roles in epithelial barrier formation. However, how claudins and other TJ membrane proteins cooperate to organize TJs remains unclear. Here, we systematically knocked out TJ components by genome editing and show that while ZO-1/ZO-2-deficient cells lacked TJ structures and epithelial barriers, claudin-deficient cells lacked TJ strands and an electrolyte permeability barrier but formed membrane appositions and a macromolecule permeability barrier. Moreover, epithelial polarity was disorganized in ZO-1/ZO-2-deficient cells, but not in claudin-deficient cells. Simultaneous deletion of claudins and a TJ membrane protein JAM-A resulted in a loss of membrane appositions and a macromolecule permeability barrier and in sporadic epithelial polarity defects. These results demonstrate that claudins and JAM-A coordinately regulate TJ formation and epithelial polarity.
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Affiliation(s)
- Tetsuhisa Otani
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan
- Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan
| | - Thanh Phuong Nguyen
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan
- Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan
| | - Shinsaku Tokuda
- Division of Nephrology and Hypertension, Department of Internal Medicine, University of Kansas Medical Center, Kansas City, KS
| | - Kei Sugihara
- Department of Anatomy and Cell Biology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan
| | - Taichi Sugawara
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan
- Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan
| | - Kyoko Furuse
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan
| | - Takashi Miura
- Department of Anatomy and Cell Biology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan
| | - Klaus Ebnet
- Institute-Associated Research Group "Cell Adhesion and Cell Polarity," Institute of Medical Biochemistry, Zentrum für Molekularbiologie der Entzündung, University of Münster, Münster, Germany
| | - Mikio Furuse
- Division of Cell Structure, National Institute for Physiological Sciences, Okazaki, Aichi, Japan
- Department of Physiological Sciences, School of Life Science, SOKENDAI (The Graduate University for Advanced Studies), Okazaki, Aichi, Japan
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Abstract
Tight junctions (TJ) play a central role in the homeostasis of epithelial and endothelial tissues, by providing a semipermeable barrier to ions and solutes, by contributing to the maintenance of cell polarity, and by functioning as signaling platforms. TJ are associated with the actomyosin and microtubule cytoskeletons, and the crosstalk with the cytoskeleton is fundamental for junction biogenesis and physiology. TJ are spatially and functionally connected to adherens junctions (AJ), which are essential for the maintenance of tissue integrity. Mechano-sensing and mechano-transduction properties of several AJ proteins have been characterized during the last decade. However, little is known about how mechanical forces act on TJ and their proteins, how TJ control the mechanical properties of cells and tissues, and what are the underlying molecular mechanisms. Here I review recent studies that have advanced our understanding of the relationships between mechanical force and TJ biology.
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Ugalde-Silva P, Navarro-Garcia F. Coordinated transient interaction of ZO-1 and afadin is required for pedestal maturation induced by EspF from enteropathogenic Escherichia coli. Microbiologyopen 2019; 8:e931. [PMID: 31568664 PMCID: PMC6925160 DOI: 10.1002/mbo3.931] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/16/2019] [Revised: 08/14/2019] [Accepted: 08/15/2019] [Indexed: 01/09/2023] Open
Abstract
Enteropathogenic Escherichia coli (EPEC) infection causes a histopathological lesion including recruitment of F‐actin beneath the attached bacteria and formation of actin‐rich pedestal‐like structures. Another important target of EPEC is the tight junction (TJ), and EspF induces displacement of TJ proteins and increased intestinal permeability. Previously, we determined that an EPEC strain lacking EspF did not cause TJ disruption; meanwhile, pedestals were located on the TJ and smaller than those induced by the wild‐type strain. Therefore, EspF could be playing an important role in both phenotypes. Here, using different cell models, we found that EspF was essential for pedestal maturation through ZO‐1 disassembly from TJ, leading to (a) ZO‐1 recruitment to the pedestal structure; no other main TJ proteins were required. Recruited ZO‐1 allowed the afadin recruitment. (b) Afadin recruitment caused an afadin–ZO‐1 transient interaction, like during TJ formation. (c) Afadin and ZO‐1 were segregated to the tip and the stem of pedestal, respectively, causing pedestal maturation. Initiation of these three discrete phases for pedestal maturation functionally and physically required EspF expression. Pedestal maturation process could help coordinate the epithelial actomyosin function by maintaining the actin‐rich column composing the pedestal structure and could be important in the dynamics of the pedestal movement on epithelial cells.
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Affiliation(s)
- Paul Ugalde-Silva
- Department of Cell Biology, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), México City, Mexico
| | - Fernando Navarro-Garcia
- Department of Cell Biology, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), México City, Mexico
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40
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Rouaud F, Vasileva E, Spadaro D, Tsukita S, Citi S. R40.76 binds to the α domain of ZO-1: role of ZO-1 (α+) in epithelial differentiation and mechano-sensing. Tissue Barriers 2019; 7:e1653748. [PMID: 31438766 PMCID: PMC6748370 DOI: 10.1080/21688370.2019.1653748] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023] Open
Abstract
The barrier function of epithelia and endothelia depends on tight junctions, which are formed by the polymerization of claudins on a scaffold of ZO proteins. Two differentially spliced isoforms of ZO-1 have been described, depending on the presence of the α domain, but the function of this domain is unclear. ZO-1 also contains a C-terminal ZU5 domain, which is involved in a mechano-sensitive intramolecular interaction with the central (ZPSG) region of ZO-1. Here we use immunoblotting and immunofluorescence to map the binding sites for commercially available monoclonal and polyclonal antibodies against ZO-1, and for a new polyclonal antibody (R3) that we developed against the ZO-1 C-terminus. We demonstrate that antibody R40.76 binds to the α domain, and the R3 antibody binds to the ZU5 domain. The (α+) isoform of ZO-1 shows higher expression in epithelial versus endothelial cells, and in differentiated versus undifferentiated primary keratinocytes, suggesting a link to epithelial differentiation and a potential molecular adaptation to junctions subjected to stronger mechanical forces. These results provide new tools and hypotheses to investigate the role of the α and ZU5 domains in ZO-1 mechano-sensing and dynamic interactions with the cytoskeleton and junctional ligands.
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Affiliation(s)
- Florian Rouaud
- Department of Cell Biology, Faculty of Sciences, University of Geneva , Geneva , Switzerland.,Institute of Genetics and Genomics of Geneva, University of Geneva , Geneva , Switzerland
| | - Ekaterina Vasileva
- Department of Cell Biology, Faculty of Sciences, University of Geneva , Geneva , Switzerland.,Institute of Genetics and Genomics of Geneva, University of Geneva , Geneva , Switzerland
| | - Domenica Spadaro
- Department of Cell Biology, Faculty of Sciences, University of Geneva , Geneva , Switzerland.,Institute of Genetics and Genomics of Geneva, University of Geneva , Geneva , Switzerland
| | - Sachiko Tsukita
- Strategic Innovation and Research Center, Teikyo University , Tokyo , Japan.,Graduate School of Frontier Biosciences, Osaka University , Osaka , Japan
| | - Sandra Citi
- Department of Cell Biology, Faculty of Sciences, University of Geneva , Geneva , Switzerland.,Institute of Genetics and Genomics of Geneva, University of Geneva , Geneva , Switzerland
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Notch-mediated inhibition of neurogenesis is required for zebrafish spinal cord morphogenesis. Sci Rep 2019; 9:9958. [PMID: 31292468 PMCID: PMC6620349 DOI: 10.1038/s41598-019-46067-1] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/15/2019] [Accepted: 06/18/2019] [Indexed: 12/26/2022] Open
Abstract
The morphogenesis of the nervous system requires coordinating the specification and differentiation of neural precursor cells, the establishment of neuroepithelial tissue architecture and the execution of specific cellular movements. How these aspects of neural development are linked is incompletely understood. Here we inactivate a major regulator of embryonic neurogenesis - the Delta/Notch pathway - and analyze the effect on zebrafish central nervous system morphogenesis. While some parts of the nervous system can establish neuroepithelial tissue architecture independently of Notch, Notch signaling is essential for spinal cord morphogenesis. In this tissue, Notch signaling is required to repress neuronal differentiation and allow thereby the emergence of neuroepithelial apico-basal polarity. Notch-mediated suppression of neurogenesis is also essential for the execution of specific morphogenetic movements of zebrafish spinal cord precursor cells. In the wild-type neural tube, cells divide at the organ midline to contribute one daughter cell to each organ half. Notch signaling deficient animals fail to display this behavior and therefore form a misproportioned spinal cord. Taken together, our findings show that Notch-mediated suppression of neurogenesis is required to allow the execution of morphogenetic programs that shape the zebrafish spinal cord.
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Christensen NR, Čalyševa J, Fernandes EFA, Lüchow S, Clemmensen LS, Haugaard‐Kedström LM, Strømgaard K. PDZ Domains as Drug Targets. ADVANCED THERAPEUTICS 2019; 2:1800143. [PMID: 32313833 PMCID: PMC7161847 DOI: 10.1002/adtp.201800143] [Citation(s) in RCA: 63] [Impact Index Per Article: 10.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/13/2018] [Revised: 03/25/2019] [Indexed: 12/14/2022]
Abstract
Protein-protein interactions within protein networks shape the human interactome, which often is promoted by specialized protein interaction modules, such as the postsynaptic density-95 (PSD-95), discs-large, zona occludens 1 (ZO-1) (PDZ) domains. PDZ domains play a role in several cellular functions, from cell-cell communication and polarization, to regulation of protein transport and protein metabolism. PDZ domain proteins are also crucial in the formation and stability of protein complexes, establishing an important bridge between extracellular stimuli detected by transmembrane receptors and intracellular responses. PDZ domains have been suggested as promising drug targets in several diseases, ranging from neurological and oncological disorders to viral infections. In this review, the authors describe structural and genetic aspects of PDZ-containing proteins and discuss the current status of the development of small-molecule and peptide modulators of PDZ domains. An overview of potential new therapeutic interventions in PDZ-mediated protein networks is also provided.
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Affiliation(s)
- Nikolaj R. Christensen
- Center for BiopharmaceuticalsDepartment of Drug Design and PharmacologyUniversity of CopenhagenUniversitetsparken 22100CopenhagenDenmark
| | - Jelena Čalyševa
- European Molecular Biology Laboratory (EMBL)Structural and Computational Biology UnitMeyerhofstraße 169117HeidelbergGermany
- EMBL International PhD ProgrammeFaculty of BiosciencesEMBL–Heidelberg UniversityGermany
| | - Eduardo F. A. Fernandes
- Center for BiopharmaceuticalsDepartment of Drug Design and PharmacologyUniversity of CopenhagenUniversitetsparken 22100CopenhagenDenmark
| | - Susanne Lüchow
- Department of Chemistry – BMCUppsala UniversityBox 576SE75123UppsalaSweden
| | - Louise S. Clemmensen
- Center for BiopharmaceuticalsDepartment of Drug Design and PharmacologyUniversity of CopenhagenUniversitetsparken 22100CopenhagenDenmark
| | - Linda M. Haugaard‐Kedström
- Center for BiopharmaceuticalsDepartment of Drug Design and PharmacologyUniversity of CopenhagenUniversitetsparken 22100CopenhagenDenmark
| | - Kristian Strømgaard
- Center for BiopharmaceuticalsDepartment of Drug Design and PharmacologyUniversity of CopenhagenUniversitetsparken 22100CopenhagenDenmark
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Valitutti F, Fasano A. Breaking Down Barriers: How Understanding Celiac Disease Pathogenesis Informed the Development of Novel Treatments. Dig Dis Sci 2019; 64:1748-1758. [PMID: 31076989 PMCID: PMC6586517 DOI: 10.1007/s10620-019-05646-y] [Citation(s) in RCA: 47] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
For decades, the pathogenesis of a variety of human diseases has been attributed to increased intestinal paracellular permeability even though scientific evidence supporting this hypothesis has been tenuous. Nevertheless, during the past decade, there have been a growing number of publications focused on human genetics, the gut microbiome, and proteomics, suggesting that loss of mucosal barrier function, particularly in the gastrointestinal tract, may substantially affect antigen trafficking, ultimately causing chronic inflammation, including autoimmunity, in genetically predisposed individuals. The gut mucosa works as a semipermeable barrier in that it permits nutrient absorption and also regulates immune surveillance while retaining potentially harmful microbes and environmental antigens within the intestinal lumen. Celiac disease (CD), a systemic, immune-mediated disorder triggered by gluten in genetically susceptible individuals, is associated with altered gut permeability. Pre-clinical and clinical studies have shown that gliadin, a prolamine component of gluten that is implicated in CD pathogenesis, is capable to disassembling intercellular junctional proteins by upregulating the zonulin pathway, which can be inhibited by the zonulin antagonist larazotide acetate. In this review, we will focus on CD as a paradigm of chronic inflammatory diseases in order to outline the contribution of gut paracellular permeability toward disease pathogenesis; moreover, we will summarize current evidence derived from available clinical trials of larazotide acetate in CD.
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Affiliation(s)
- Francesco Valitutti
- Pediatric Gastroenterology and Liver Unit, Department of "Maternal-and-Child Health" and Urology, Sapienza University of Rome, Rome, Italy
| | - Alessio Fasano
- Mucosal Immunology and Biology Research Center, Center for Celiac Research and Treatment and Division of Pediatric Gastroenterology and Nutrition, MassGeneral Hospital for Children, 175 Cambridge Street, CPZS - 574, Boston, MA, 02114, USA.
- European Biomedical Research Institute of Salerno, Salerno, Italy.
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Abstract
The term blood-bile barrier (BBlB) refers to the physical structure within a hepatic lobule that compartmentalizes and hence segregates sinusoidal blood from canalicular bile. Thus, this barrier provides physiological protection in the liver, shielding the hepatocytes from bile toxicity and restricting the mixing of blood and bile. BBlB is primarily composed of tight junctions; however, adherens junction, desmosomes, gap junctions, and hepatocyte bile transporters also contribute to the barrier function of the BBlB. Recent findings also suggest that disruption of BBlB is associated with major hepatic diseases characterized by cholestasis and aberrations in BBlB thus may be a hallmark of many chronic liver diseases. Several molecular signaling pathways have now been shown to play a role in regulating the structure and function and eventually contribute to regulation of the BBlB function within the liver. In this review, we will discuss the structure and function of the BBlB, summarize the methods to assess the integrity and function of BBlB, discuss the role of BBlB in liver pathophysiology, and finally, discuss the mechanisms of BBlB regulation. Collectively, this review will demonstrate the significance of the BBlB in both liver homeostasis and hepatic dysfunction.
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Affiliation(s)
- Tirthadipa Pradhan-Sundd
- *Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
- †Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
| | - Satdarshan Pal Monga
- *Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
- †Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
- ‡Pittsburgh Liver Research Center, University of Pittsburgh School of Medicine and University of Pittsburgh Medical Center, Pittsburgh, PA, USA
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Restoring retinal neurovascular health via substance P. Exp Cell Res 2019; 380:115-123. [PMID: 30995434 PMCID: PMC6548993 DOI: 10.1016/j.yexcr.2019.04.008] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/19/2019] [Revised: 03/28/2019] [Accepted: 04/05/2019] [Indexed: 12/24/2022]
Abstract
Regulation of vascular permeability plays a major role in the pathophysiology of visually threatening conditions such as retinal vein occlusion and diabetic retinopathy. Principally, several factors such as vascular endothelial growth factor (VEGF), are up-regulated or induced in response to hypoxia thus adversely affecting the blood-retinal barrier (BRB), resulting in retinal edema and neovascularisation. Furthermore, current evidence supports a dysregulation of the inner retinal neural-vascular integrity as a critical factor driving retinal ganglion cell (RGC) death and visual loss. The principal objective of this study was to interrogate whether Substance P (SP), a constitutive neurotransmitter of amacrine and ganglion cells, may protect against N-methyl-d-aspartate (NMDA)-induced excitotoxic apoptosis of ganglion cells and VEGF-induced vessel leakage in the retina. Tight junctional protein expression and a Vascular Permeability Image Assay were used to determine vascular integrity in vitro. The protective effect of SP on RGC was established in ex vivo retinal explants and in vivo murine models. After NMDA administration, a reduction in TUNEL+ cells and a maintained number of Brn-3a+ cells were found, indicating an inhibition of RGC apoptosis mediated by SP. Additionally, SP maintained endothelial tight junctions and decreased VEGF-induced vascular permeability. In conclusion, administration of SP protects against NMDA apoptosis of RGC and VEGF-induced endothelial barrier breakdown.
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Bloch S, Thomas M, Colin I, Galant S, Machado E, Affaticati P, Jenett A, Yamamoto K. Mesencephalic origin of the inferior lobe in zebrafish. BMC Biol 2019; 17:22. [PMID: 30849972 PMCID: PMC6407210 DOI: 10.1186/s12915-019-0631-y] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/10/2018] [Accepted: 01/22/2019] [Indexed: 11/18/2022] Open
Abstract
BACKGROUND Although the overall brain organization is shared in vertebrates, there are significant differences within subregions among different groups, notably between Sarcopterygii (lobe-finned fish) and Actinopterygii (ray-finned fish). Recent comparative studies focusing on the ventricular morphology have revealed a large diversity of the hypothalamus. Here, we study the development of the inferior lobe (IL), a prominent structure forming a bump on the ventral surface of the teleost brain. Based on its position, IL has been thought to be part of the hypothalamus (therefore forebrain). RESULTS Taking advantage of genetic lineage-tracing techniques in zebrafish, we reveal that cells originating from her5-expressing progenitors in the midbrain-hindbrain boundary (MHB) participate in the formation of a large part of the IL. 3D visualization demonstrated how IL develops in relation to the ventricular system. We found that IL is constituted by two developmental components: the periventricular zone of hypothalamic origin and the external zone of mesencephalic origin. The mesencephalic external zone grows progressively until adulthood by adding new cells throughout development. CONCLUSION Our results disprove a homology between the IL and the mammalian lateral hypothalamus. We suggest that the IL is likely to be involved in multimodal sensory integration rather than feeding motivation. The teleost brain is not a simpler version of the mammalian brain, and our study highlights the evolutionary plasticity of the brain which gives rise to novel structures.
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Affiliation(s)
- Solal Bloch
- Paris-Saclay Institute of Neuroscience (Neuro-PSI), CNRS UMR9197, Univ Paris Sud, Université Paris-Saclay, CNRS Bâtiment 5, Avenue de la Terrasse, 91190, Gif-sur-Yvette, France
| | - Manon Thomas
- Paris-Saclay Institute of Neuroscience (Neuro-PSI), CNRS UMR9197, Univ Paris Sud, Université Paris-Saclay, CNRS Bâtiment 5, Avenue de la Terrasse, 91190, Gif-sur-Yvette, France
- Present address: Plateau de phénotypage TEFOR, LPGP-INRA UR1037, 35042, Rennes, France
| | - Ingrid Colin
- Paris-Saclay Institute of Neuroscience (Neuro-PSI), CNRS UMR9197, Univ Paris Sud, Université Paris-Saclay, CNRS Bâtiment 5, Avenue de la Terrasse, 91190, Gif-sur-Yvette, France
| | - Sonya Galant
- Paris-Saclay Institute of Neuroscience (Neuro-PSI), CNRS UMR9197, Univ Paris Sud, Université Paris-Saclay, CNRS Bâtiment 5, Avenue de la Terrasse, 91190, Gif-sur-Yvette, France
| | - Elodie Machado
- TEFOR Paris-Saclay, CNRS UMS2010, INRA UMS1451, Univ Paris Sud, Université Paris-Saclay, 91190, Gif-sur-Yvette, France
| | - Pierre Affaticati
- TEFOR Paris-Saclay, CNRS UMS2010, INRA UMS1451, Univ Paris Sud, Université Paris-Saclay, 91190, Gif-sur-Yvette, France
| | - Arnim Jenett
- TEFOR Paris-Saclay, CNRS UMS2010, INRA UMS1451, Univ Paris Sud, Université Paris-Saclay, 91190, Gif-sur-Yvette, France
| | - Kei Yamamoto
- Paris-Saclay Institute of Neuroscience (Neuro-PSI), CNRS UMR9197, Univ Paris Sud, Université Paris-Saclay, CNRS Bâtiment 5, Avenue de la Terrasse, 91190, Gif-sur-Yvette, France.
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Liu X, Fuentes EJ. Emerging Themes in PDZ Domain Signaling: Structure, Function, and Inhibition. INTERNATIONAL REVIEW OF CELL AND MOLECULAR BIOLOGY 2019; 343:129-218. [PMID: 30712672 PMCID: PMC7185565 DOI: 10.1016/bs.ircmb.2018.05.013] [Citation(s) in RCA: 37] [Impact Index Per Article: 6.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 02/02/2023]
Abstract
Post-synaptic density-95, disks-large and zonula occludens-1 (PDZ) domains are small globular protein-protein interaction domains widely conserved from yeast to humans. They are composed of ∼90 amino acids and form a classical two α-helical/six β-strand structure. The prototypical ligand is the C-terminus of partner proteins; however, they also bind internal peptide sequences. Recent findings indicate that PDZ domains also bind phosphatidylinositides and cholesterol. Through their ligand interactions, PDZ domain proteins are critical for cellular trafficking and the surface retention of various ion channels. In addition, PDZ proteins are essential for neuronal signaling, memory, and learning. PDZ proteins also contribute to cytoskeletal dynamics by mediating interactions critical for maintaining cell-cell junctions, cell polarity, and cell migration. Given their important biological roles, it is not surprising that their dysfunction can lead to multiple disease states. As such, PDZ domain-containing proteins have emerged as potential targets for the development of small molecular inhibitors as therapeutic agents. Recent data suggest that the critical binding function of PDZ domains in cell signaling is more than just glue, and their binding function can be regulated by phosphorylation or allosterically by other binding partners. These studies also provide a wealth of structural and biophysical data that are beginning to reveal the physical features that endow this small modular domain with a central role in cell signaling.
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Affiliation(s)
- Xu Liu
- Department of Biochemistry, University of Iowa, Iowa City, IA, United States
| | - Ernesto J. Fuentes
- Department of Biochemistry, University of Iowa, Iowa City, IA, United States
- Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA, United States
- Corresponding author: E-mail:
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Yin J, Lv L, Zhai P, Long T, Zhou Q, Pan H, Botwe G, Wang L, Wang Q, Tan L, Kuebler WM. Connexin 40 regulates lung endothelial permeability in acute lung injury via the ROCK1-MYPT1- MLC20 pathway. Am J Physiol Lung Cell Mol Physiol 2019; 316:L35-L44. [PMID: 30234377 DOI: 10.1152/ajplung.00012.2018] [Citation(s) in RCA: 24] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
Increased pulmonary vascular permeability is a hallmark of acute lung injury (ALI). Connexin 40 (Cx40) is a gap junctional protein abundantly present in the lung microvascular endothelium. Yet, the role of Cx40 in the regulation of lung vascular permeability and its underlying mechanisms are unclear. Here, we tested the hypothesis that Cx40 participates in regulation of lung endothelial permeability via a mechanism involving a Rho-associated protein kinase (ROCK) dependent regulation of myosin light chain (MLC). In murine models of intratracheal acid- or LPS-induced lung injury, genetic deficiency of Cx40 attenuated key features of ALI including vascular barrier failure. In human pulmonary microvascular endothelial cells (PMVECs), thrombin-induced loss of transendothelial electrical resistance was attenuated by a Cx40-inhibiting mimetic peptide (40GAP27), Cx40-specific shRNA, or ROCK inhibitor Y27632. In isolated perfused mouse lungs, platelet-activating factor-induced lung weight gain was abrogated by gap junction blocker carbenoxolone, 40GAP27, Y27632, or genetic deficiency of Cx40. Phosphorylation of MLC20 increased drastically in both LPS-treated PMVECs and HCl-treated mouse lungs. Expression of ROCK1 was increased in both LPS-treated PMVECs and HCl-treated mouse lungs, and paralleled by phosphorylation of MLC20. Coimmunoprecipitation experiments revealed protein-protein interaction between ROCK1 and Cx40. LPS-induced upregulation of ROCK1 and phosphorylation of MLC20 were blocked by knockdown of Cx40. LPS caused phosphorylation of myosin phosphatase targeting subunit 1, which could be abrogated by Y27632 or Cx40-shRNA. Our findings reveal a role of Cx40 in regulation of ROCK1 and MLC20 that contributes critically to lung vascular barrier failure in ALI.
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Affiliation(s)
- Jun Yin
- Department of Thoracic Surgery, Zhongshan Hospital of Fudan University , Shanghai , China
- Department of Cardiothoracic Surgery, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Lu Lv
- Department of Cardiothoracic Surgery, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Peng Zhai
- Department of Cardiothoracic Surgery, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Tao Long
- Department of Cardiothoracic Surgery, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Qiang Zhou
- Department of Cardiothoracic Surgery, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Huiwen Pan
- Department of Cardiothoracic Surgery, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Godwin Botwe
- Department of Cardiothoracic Surgery, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Liming Wang
- Department of Chemotherapy, Cancer Institute, Affiliated People's Hospital of Jiangsu University , Zhenjiang, Jiangsu , China
| | - Qun Wang
- Department of Thoracic Surgery, Zhongshan Hospital of Fudan University , Shanghai , China
| | - Lijie Tan
- Department of Thoracic Surgery, Zhongshan Hospital of Fudan University , Shanghai , China
| | - Wolfgang M Kuebler
- Department of Physiology and Center for Cardiovascular Research, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health , Berlin , Germany
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Sharma UC, Sonkawade SD, Baird A, Chen M, Xu S, Sexton S, Singh AK, Groman A, Turowski SG, Spernyak JA, Mahajan SD, Pokharel S. Effects of a novel peptide Ac-SDKP in radiation-induced coronary endothelial damage and resting myocardial blood flow. CARDIO-ONCOLOGY (LONDON, ENGLAND) 2018; 4:8. [PMID: 31057947 PMCID: PMC6497419 DOI: 10.1186/s40959-018-0034-1] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 09/21/2018] [Accepted: 11/28/2018] [Indexed: 12/15/2022]
Abstract
BACKGROUND Cancer survivors treated with thoracic ionizing radiation are at higher risk of premature death due to myocardial ischemia. No therapy is currently available to prevent or mitigate these effects. We tested the hypothesis that an endogenous tetrapeptide N-acetyl-Ser-Asp-Lys-Pro (Ac-SDKP) counteracts radiation-induced coronary vascular fibrosis and endothelial cell loss and preserves myocardial blood flow. METHODS We examined a rat model with external-beam-radiation exposure to the cardiac silhouette. We treated a subgroup of irradiated rats with subcutaneous Ac-SDKP for 18-weeks. We performed cardiac MRI with Gadolinium contrast to examine resting myocardial blood flow content. Upon sacrifice, we examined coronary endothelial-cell-density, fibrosis, apoptosis and endothelial tight-junction proteins (TJP). In vitro, we examined Ac-SDKP uptake by the endothelial cells and tested its effects on radiation-induced reactive oxygen species (ROS) generation. In vivo, we injected labeled Ac-SDKP intravenously and examined its endothelial localization after 4-h. RESULTS We found that radiation exposure led to reduced resting myocardial blood flow content. There was concomitant endothelial cell loss and coronary fibrosis. Smaller vessels and capillaries showed more severe changes than larger vessels. Real-time PCR and confocal microscopy showed radiation-induced loss of TJ proteins including-claudin-1 and junctional adhesion molecule-2 (JAM-2). Ac-SDKP normalized myocardial blood flow content, inhibited endothelial cell loss, reduced coronary fibrosis and restored TJ-assembly. In vitro, Ac-SDKP localized to endothelial cells and inhibited radiation-induced endothelial ROS generation. In vivo, labeled Ac-SDKP was visualized into the endothelium 4-h after the intravenous injection. CONCLUSIONS We concluded that Ac-SDKP has protective effects against radiation-induced reduction of myocardial blood flow. Such protective effects are likely mediated by neutralization of ROS-mediated injury, preservation of endothelial integrity and inhibition of fibrosis. This demonstrates a strong therapeutic potential of Ac-SDKP to counteract radiotherapy-induced coronary disease.
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Affiliation(s)
- Umesh C. Sharma
- Department of Medicine, Division of Cardiology, Jacob’s School of Medicine and Biomedical Sciences, Buffalo, NY USA
| | - Swati D. Sonkawade
- Department of Medicine, Division of Cardiology, Jacob’s School of Medicine and Biomedical Sciences, Buffalo, NY USA
| | - Andrew Baird
- Department of Medicine, Division of Cardiology, Jacob’s School of Medicine and Biomedical Sciences, Buffalo, NY USA
| | - Min Chen
- Department of Pathology, Division of Thoracic Pathology and Oncology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203 USA
| | - Shirley Xu
- Department of Medicine, Division of Cardiology, Jacob’s School of Medicine and Biomedical Sciences, Buffalo, NY USA
- Department of Pathology, Division of Thoracic Pathology and Oncology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203 USA
| | - Sandra Sexton
- Laboratory Animal Shared Resource Facility, Roswell Park Cancer Center, Buffalo, NY USA
| | - Anurag K. Singh
- Department of Radiation Oncology, Roswell Park Comprehensive Cancer Center, Buffalo, NY USA
| | - Adrienne Groman
- Department of Biostatistics and Bioinformatics, Roswell Park Comprehensive Cancer Center, Buffalo, NY USA
| | - Steven G. Turowski
- Translational Imaging Shared Resources, Roswell Park Comprehensive Cancer Center, Buffalo, NY USA
| | - Joseph A. Spernyak
- Translational Imaging Shared Resources, Roswell Park Comprehensive Cancer Center, Buffalo, NY USA
| | - Supriya D. Mahajan
- Department of Medicine, Jacob’s School of Medicine and Biomedical Sciences, Buffalo, NY USA
| | - Saraswati Pokharel
- Department of Pathology, Division of Thoracic Pathology and Oncology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203 USA
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Rhee SW, Rusch NJ. Molecular determinants of beta-adrenergic signaling to voltage-gated K + channels in the cerebral circulation. Microcirculation 2018; 25. [PMID: 29072364 DOI: 10.1111/micc.12425] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/11/2017] [Accepted: 10/19/2017] [Indexed: 12/14/2022]
Abstract
Voltage-gated K+ (Kv ) channels are major determinants of membrane potential in vascular smooth muscle cells (VSMCs) and regulate the diameter of small cerebral arteries and arterioles. However, the intracellular structures that govern the expression and function of vascular Kv channels are poorly understood. Scaffolding proteins including postsynaptic density 95 (PSD95) recently were identified in rat cerebral VSMCs. Primarily characterized in neurons, the PSD95 scaffold has more than 50 known binding partners, and it can mediate macromolecular signaling between cell-surface receptors and ion channels. In cerebral arteries, Shaker-type Kv 1 channels appear to associate with the PSD95 molecular scaffold, and PSD95 is required for the normal expression and vasodilator influence of members of this K+ channel gene family. Furthermore, recent findings suggest that the β1-subtype adrenergic receptor is expressed in cerebral VSMCs and forms a functional vasodilator complex with Kv 1 channels on the PSD95 scaffold. Activation of β1-subtype adrenergic receptors in VSMCs enables protein kinase A-dependent phosphorylation and opening of Kv 1 channels in the PSD95 complex; the subsequent K+ efflux mediates membrane hyperpolarization and vasodilation of small cerebral arteries. Early evidence from other studies suggests that other families of Kv channels and scaffolding proteins are expressed in VSMCs. Future investigations into these macromolecular complexes that modulate the expression and function of Kv channels may reveal unknown signaling cascades that regulate VSMC excitability and provide novel targets for ion channel-based medications to optimize vascular tone.
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Affiliation(s)
- Sung W Rhee
- Department of Pharmacology and Toxicology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, USA
| | - Nancy J Rusch
- Department of Pharmacology and Toxicology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, USA
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