|
1
|
Hernández-Rodríguez M, Arciniega-Martínez IM, García-Marín ID, Correa-Basurto J, Rosales-Hernández MC. Chronic Administration of Scopolamine Increased GSK3βP9, Beta Secretase, Amyloid Beta, and Oxidative Stress in the Hippocampus of Wistar Rats. Mol Neurobiol 2020; 57:3979-3988. [PMID: 32638218 DOI: 10.1007/s12035-020-02009-x] [Citation(s) in RCA: 21] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/04/2020] [Accepted: 07/01/2020] [Indexed: 10/23/2022]
Abstract
The increase of amyloid beta (Aβ) release and hyperphosphorylation of Tau protein represents the main events related to Alzheimer's disease (AD). Furthermore, the sporadic type represents the most common form of AD. Therefore, the establishment of a non-transgenic animal model that resembles the characteristics of the disease is of particular importance. Scopolamine has been linked to increases in both Aβ production and oxidative stress in rat and mice brains. Thus, the purpose of the present work was to identify changes in biomarkers that are related to AD after chronic administration of scopolamine (2 mg/kg i.p., during 6 and 12 weeks) to male Wistar rats. The results showed increased Aβ deposition at rat hippocampus which could be due to an increase of β-site amyloid-β-protein precursor cleaving enzyme 1 (BACE1) expression and activity. These findings could be related to the increase of glycogen synthase kinase 3 phosphorylated (GSK3βP9) expression. Finally, the establishment of a state of oxidative stress in groups treated with scopolamine was demonstrated by an increase in free radical content and MDA levels. The present study facilitates our understanding of the changes that occur in biomolecules related to AD in Wistar rats after the chronic administration of scopolamine.
Collapse
Affiliation(s)
- Maricarmen Hernández-Rodríguez
- Laboratorio de Modelado Molecular y Bioinformática, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, Mexico.,Laboratorio de Biofísica y Biocatálisis, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, Mexico
| | - Ivonne Maciel Arciniega-Martínez
- Laboratorio de Inmunidad de Mucosas, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, Mexico
| | - Iohanan Daniel García-Marín
- Laboratorio de Modelado Molecular y Bioinformática, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, Mexico.,Laboratorio de Biofísica y Biocatálisis, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, Mexico
| | - José Correa-Basurto
- Laboratorio de Modelado Molecular y Bioinformática, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, Mexico.
| | - Martha Cecilia Rosales-Hernández
- Laboratorio de Biofísica y Biocatálisis, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón s/n, 11340, Mexico, Mexico.
| |
Collapse
|
|
2
|
Doganavsargil B, Buberal GE, Toz H, Sarsik B, Pehlivanoglu B, Sezak M, Sen S. Digitally reinforced hematoxylin-eosin polarization technique in diagnosis of rectal amyloidosis. World J Gastroenterol 2015; 21:1827-1837. [PMID: 25684948 PMCID: PMC4323459 DOI: 10.3748/wjg.v21.i6.1827] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/05/2014] [Revised: 11/28/2014] [Accepted: 12/22/2014] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate the efficacy of the digitally reinforced hematoxylin-eosin polarization (DRHEP) technique for detection of amyloidosis in rectal biopsies.
METHODS: One hundred hematoxylin-eosin (HE) stained rectal biopsies with Congo-red (CR)-positive amyloid depositions and 50 control cases with CR-negative amyloid-mimicking areas were scanned blinded to the CR results for amyloid depositions under both bright and polarized light, and digitally photographed using the DRHEP technique, to accentuate the faint birefringence observed in HE slides under polarization. The results of DRHEP and HE evaluation were statistically correlated with CR polarization results with respect to presence and localization of amyloid deposits as well as amyloid types.
RESULTS: Amyloid deposits showed yellowish-green birefringence by DRHEP, which allowed identification of amyloidosis in 41 HE-unsuspected cases (P = 0.016), 31 of which only had vascular deposits. True positivity was higher, and false negativity and positivity were lower by DRHEP, compared to evaluation by HE (69%, 31%, and 0.8% vs 33%, 67%, and 33%, respectively; P < 0.0001). The sensitivity, specificity, accuracy, and positive and negative predictive values for DRHEP were 69%, 98%, 78.6%, 98.5%, and 61.25%, respectively. Reasons for DRHEP false negativity were presence of extensive background birefringence in 12 cases, absence of CR birefringent vessel in 3 cases, and missing of the tiny deposits in 9 cases, which could be improved by experience, especially in the latter case. No correlation was found between age, gender, sites of deposits, or amyloid types.
CONCLUSION: The DRHEP technique improves diagnostic accuracy when used as an adjunct or a prior step to CR staining, especially for cases with limited tissues for further analysis.
Collapse
|
|
3
|
Fluorescence Investigation of Interactions Between Novel Benzanthrone Dyes and Lysozyme Amyloid Fibrils. J Fluoresc 2013; 24:493-504. [DOI: 10.1007/s10895-013-1318-3] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/22/2013] [Accepted: 11/07/2013] [Indexed: 10/25/2022]
|
|
4
|
Csoka L, Appel TR, Eitner A, Jirikowski G, Makovitzky J. Polarization optical-histochemical characterization and supramolecular structure of carbohydrate fibrils. Acta Histochem 2013; 115:22-31. [PMID: 22497931 DOI: 10.1016/j.acthis.2012.03.003] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/04/2012] [Revised: 03/13/2012] [Accepted: 03/15/2012] [Indexed: 11/23/2022]
Abstract
Topo-optical staining reactions were used to investigate the structures of bacterial cellulose, insect chitosan and alginic acid from brown algae. Polysaccharide complexes, glycosaminoglycans and sulfate groups were presented and demonstrated selectively. Chitosan and alginic acid are structurally similar to glycosaminoglycans (GAGs), which are constituents of human amyloid fibrils. The staining sequences shown can be used as reliable methods for histochemistry with light and polarization microscopy. They will help to clarify the complex protein-polysaccharide structure of amyloid fibrils.
Collapse
Affiliation(s)
- Levente Csoka
- University of West Hungary, Institute of Wood and Paper Technology, Sopron, Hungary.
| | | | | | | | | |
Collapse
|
|
5
|
Jensen JR, Cisek K, Honson NS, Kuret J. Ligand polarizability contributes to tau fibril binding affinity. Bioorg Med Chem 2011; 19:5147-54. [PMID: 21803586 DOI: 10.1016/j.bmc.2011.07.016] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/25/2011] [Revised: 07/07/2011] [Accepted: 07/11/2011] [Indexed: 10/18/2022]
Abstract
Whole brain imaging of tau-bearing neurofibrillary lesions has the potential to improve the premortem diagnosis and staging of Alzheimer's disease. Diverse compounds with high affinity for tau aggregates have been reported from high-throughput screens, but the affinity driving features common among them have not been determined. To identify these features, analogs of compounds discovered by high-throughput screening, including phenothiazine, triarylmethine, benzothiazole, and oxindole derivatives, were tested for their ability to displace fluorescent thioflavin dyes from filaments made from recombinant tau protein or authentic paired helical filaments purified from Alzheimer's disease tissue. When representative members of all scaffolds were assayed, the rank order of binding affinity determined for synthetic and authentic filaments correlated strongly, indicating that synthetic filaments have predictive utility for ligand development. Within individual scaffold families, binding affinity was found to correlate with compound polarizability, consistent with a role for dispersion forces in mediating ligand binding. Overall, the data indicate that polarizability is an important commonality among structurally diverse tau binding ligands, and that affinity for tau aggregates can be maximized by integrating formal assessment of this parameter into ligand discovery efforts.
Collapse
Affiliation(s)
- Jordan R Jensen
- Department of Molecular and Cellular Biochemistry, The Ohio State University College of Medicine, Columbus, OH 43210, USA
| | | | | | | |
Collapse
|
|
6
|
Howie AJ, Owen-Casey MP. Discrepancies between descriptions and illustrations of colours in Congo red-stained amyloid, and explanation of discrepant colours. Amyloid 2010; 17:109-17. [PMID: 21039325 DOI: 10.3109/13506129.2010.527448] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
Abstract
Congo red-stained sections of amyloid may show various colours between crossed polariser and analyser. The aims were to see how papers described the colours, to compare descriptions with illustrations, and to explain the colours. In 160 papers on Congo red-stained amyloid, the commonest descriptions were 'green birefringence' and 'apple-green birefringence'. In 191 figures in 82 papers, 59 (31%) showed a pure green colour, 62 (32%) showed green and yellow or blue and yellow, 38 (20%) showed green and a colour other than yellow, mostly red, and 32 (17%) showed other colours. Discrepancies between colours reported and illustrated were noted in 127 figures (66%). Most (77) were between green alone in descriptions and green and another colour in figures, and 30 were between green in descriptions and no green at all in figures. Pure green can be seen in ideal conditions, but more often there are green and yellow, explained by strain birefringence, and green and red or other combinations, explained by uncrossing of polariser and analyser. These other anomalous colours are just as characteristic of amyloid as the pure green colour. Many papers on Congo red-stained amyloid appear to describe what is expected theoretically rather than what is actually seen.
Collapse
|
|
7
|
Steinbach G, Pomozi I, Jánosa DP, Makovitzky J, Garab G. Confocal fluorescence detected linear dichroism imaging of isolated human amyloid fibrils. Role of supercoiling. J Fluoresc 2010; 21:983-9. [PMID: 20556489 DOI: 10.1007/s10895-010-0684-3] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/05/2010] [Accepted: 06/01/2010] [Indexed: 10/19/2022]
Abstract
Amyloids are highly organized insoluble protein aggregates that are associated with a large variety of degenerative diseases. In this work, we investigated the anisotropic architecture of isolated human amyloid samples stained with Congo Red. This was performed by fluorescence detected linear dichroism (FDLD) imaging in a laser scanning confocal microscope that was equipped with a differential polarization attachment using high frequency modulation of the polarization state of the laser beam and a demodulation circuit. Two- and three-dimensional FDLD images of amyloids provided information on the orientation of the electric transition dipoles of the intercalated Congo Red molecules with unprecedented precision and spatial resolution. We show that, in accordance with linear dichroism imaging (Jin et al. Proc Natl Acad Sci USA 100:15294, 2003), amyloids exhibit strong anisotropy with preferential orientation of the dye molecules along the fibrils; estimations on the orientation angle, of around 45°, are given using a model calculation which takes into account the helical organization of the filaments and fibrils. Our data also show that FDLD images display large inhomogeneities, high local values with alternating signs and, in some regions, well identifiable µm-sized periodicities. These features of the anisotropic architecture are accounted for by supercoiling of helically organized amyloid fibrils.
Collapse
Affiliation(s)
- Gábor Steinbach
- Biological Research Center, Hungarian Academy of Sciences, P.O. Box 521, Szeged 6701, Hungary
| | | | | | | | | |
Collapse
|
|
8
|
Makovitzky J, Richter S. The relevance of the aldehyde bisulfite toluidine blue reaction and its variants in the submicroscopic carbohydrate research. Acta Histochem 2009; 111:273-91. [PMID: 19157525 PMCID: PMC7172417 DOI: 10.1016/j.acthis.2008.11.027] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Carbohydrates are chemical compounds that contain only oxygen, hydrogen and carbon. They are classified by their number of sugar units: monosaccharides (such as glucose and fructose), and disaccharides (such as sucrose and lactose) are simple carbohydrates; oligosaccharides and polysaccharides (such as starch, glycogen and cellulose) are complex carbohydrates. Carbohydrates play a crucial role in diverse biological systems [Hricovín M. Structural aspects of carbohydrates and the relation with their biological properties. Curr Med Chem 2004;11:2565-83]. According to Roseman [Sugars of the cell membrane. In: Weissmann G, Clairborn E, editors. Cell membranes. Biochemistry, Cell Biology, Pathology. New York: H. P. Publ. Co; 1975. p. 55-64], two classes of glycoproteins are described. Free glycoproteins are localised in the surface coat of the membranes and form a thick mobile layer, without any association to the membrane itself. Functionally, however, they are located in a close association with the membrane (e.g. in the duodenal mucosa). The other group consists of the membrane glycoproteins, which are integral to the membranes and are located in the outer layer. The oligosaccharide chains are bound to the N-terminal part of proteins, and are situated in the hydrophilic zone. Glycoproteins have diverse functions. They are important in specific receptor functions, in immunological cell destruction and play a significant role in reactions with lectins, antibodies, as well as in cell association and mutual recognition of the cells. This paper focuses on aspects of a summary of polarisation optical investigations and biological functions of the following three groups of carbohydrates: oligosaccharides, glycoproteins and glycosaminoglycans.
Collapse
Affiliation(s)
- Josef Makovitzky
- Department of Neuropathology, University Heidelberg, Im Neuenheimer Feld 220, D-69120 Heidelberg, Germany.
| | | |
Collapse
|
|
9
|
House E, Mold M, Collingwood J, Baldwin A, Goodwin S, Exley C. Copper abolishes the beta-sheet secondary structure of preformed amyloid fibrils of amyloid-beta(42). J Alzheimers Dis 2009; 18:811-7. [PMID: 19749401 PMCID: PMC2857508 DOI: 10.3233/jad-2009-1235] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
Abstract
The observation of the co-deposition of metals and amyloid-beta(42) (Abeta(42)) in brain tissue in Alzheimer's disease prompted myriad investigations into the role played by metals in the precipitation of this peptide. Copper is bound by monomeric Abeta(12) and upon precipitation of the copper-peptide complex thereby prevents Abeta(42) from adopting a beta-sheet secondary structure. Copper is also bound by beta-sheet conformers of Abeta(42), and herein we have investigated how this interaction affects the conformation of the precipitated peptide. Copper significantly reduced the thioflavin T fluorescence of aged, fibrillar Abeta(42) with, for example, a 20-fold excess of the metal resulting in a ca 90% reduction in thioflavin T fluorescence. Transmission electron microscopy showed that copper significantly reduced the quantities of amyloid fibrils while Congo red staining and polarized light demonstrated a copper-induced abolition of apple-green birefringence. Microscopy under cross-polarized light also revealed the first observation of spherulites of Abeta(42). The size and appearance of these amyloid structures were found to be very similar to spherulites identified in Alzheimer's disease tissue. The combined results of these complementary methods strongly suggested that copper abolished the beta-sheet secondary structure of pre-formed, aged amyloid fibrils of Abeta(42). Copper may protect against the presence of beta-sheets of Abeta(42) in vivo, and its binding by fibrillar Abeta(42) could have implications for Alzheimer's disease therapy.
Collapse
Affiliation(s)
- Emily House
- The Birchall Centre, Lennard-Jones Laboratories, Keele University, Staffordshire, UK
| | - Matthew Mold
- The Birchall Centre, Lennard-Jones Laboratories, Keele University, Staffordshire, UK
| | | | - Alex Baldwin
- Life Sciences, Keele University, Staffordshire, UK
| | | | - Christopher Exley
- The Birchall Centre, Lennard-Jones Laboratories, Keele University, Staffordshire, UK
| |
Collapse
|
|
10
|
Richter S, Makovitzky J. Topo-optical visualization reactions of carbohydrate-containing amyloid deposits in the respiratory tract. Acta Histochem 2006; 108:181-91. [PMID: 16542712 DOI: 10.1016/j.acthis.2006.01.002] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/20/2005] [Revised: 12/15/2005] [Accepted: 01/11/2006] [Indexed: 11/20/2022]
Abstract
Staining with Congo red according to is the most commonly used method for the demonstration of amyloid, but structures other than amyloid can give false-positive results. To overcome this problem, introduced an aqueous Congo red staining with gum arabic as the mounting medium, which we have used in this and previous publications. Most histochemical studies on amyloid deposits to date have concentrated on conventional methods including staining with thioflavine, sirius red, alcian blue, methyl and crystal violet. In this study, we used topo-optical reactions with thiazine dyes on both the light and polarization microscopic level to establish the structure, distribution and location of carbohydrate components that occur within amyloid deposits, especially in the respiratory tract. Topo-optical staining reactions for the qualitative analysis of carbohydrate components in amyloid deposits included (1) reactions that identify the carbohydrate residues, (2) reactions that detect sialic acids and, (3) methods that visualize glycosaminoglycans. In conclusion, a comparison of consecutive serial sections stained with Congo red, aldehyde bisulfite toluidine blue reaction, sialic acid-specific topo-optical reaction, toluidine blue topo-optical reaction and chemically intensified basophilic reaction showed correlative staining patterns and anisotropic effects, corresponding to a close pathomorphological relationship between amyloid fibrils, periodate reactive carbohydrates, including sialic acids, and glycosaminoglycans.
Collapse
Affiliation(s)
- Susann Richter
- Institute of Pathology, Clinic of Gynaecology and Obstetrics, University of Rostock, Südring 81, 18059 Rostock, Germany
| | | |
Collapse
|