Basic Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hematol. Nov 6, 2016; 5(4): 88-93
Published online Nov 6, 2016. doi: 10.5315/wjh.v5.i4.88
P2X7 receptor activation causes phosphatidylserine exposure in canine erythrocytes
Megan Faulks, Tracey A Kuit, Reece A Sophocleous, Belinda L Curtis, Stephen J Curtis, Lisa M Jurak, Ronald Sluyter
Megan Faulks, Tracey A Kuit, Reece A Sophocleous, Lisa M Jurak, Ronald Sluyter, School of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, Australia
Megan Faulks, Reece A Sophocleous, Lisa M Jurak, Ronald Sluyter, Centre for Medical and Molecular Bioscience, University of Wollongong, Wollongong, NSW 2522, Australia
Megan Faulks, Reece A Sophocleous, Lisa M Jurak, Ronald Sluyter, Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia
Belinda L Curtis, Stephen J Curtis, Albion Park Veterinary Hospital, Albion Park, NSW 2527, Australia
Author contributions: Kuit TA and Sluyter R designed the research; Faulks M, Sophocleous RA and Jurak LM performed the research; Faulks M, Sophocleous RA and Sluyter R analysed the data; Curtis BL and Curtis SJ contributed samples; Sluyter R wrote the paper; Faulks M, Sophocleous RA, Curtis BL, Curtis SJ and Jurak LM reviewed the paper.
Supported by The Centre for Medical and Molecular Bioscience (University of Wollongong); and the American Kennel Club Canine Health Foundation.
Institutional review board statement: The study was reviewed and approved by the Human and Animal Ethics Committees of the University of Wollongong.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Animal Ethics Committee of the University of Wollongong (Protocol AE14/09). Consent from pet owners was reviewed and approved by the Human Ethics Committee of the University of Wollongong (Protocol HE10/063).
Conflict-of-interest statement: The authors declare no conflicts of interest regarding this manuscript.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Ronald Sluyter, PhD, Associate Professor, School of Biological Sciences, University of Wollongong, Northfields Ave, Wollongong, NSW 2522, Australia. rsluyter@uow.edu.au
Telephone: +61-2-42215508 Fax: +61-2-42218130
Received: July 20, 2016
Peer-review started: July 21, 2016
First decision: August 5, 2016
Revised: August 31, 2016
Accepted: September 21, 2016
Article in press: September 23, 2016
Published online: November 6, 2016
Processing time: 106 Days and 1.4 Hours
Abstract
AIM

To determine if activation of the ATP-gated P2X7 receptor channel induces phosphatidylserine (PS) exposure in erythrocytes from multiple dog breeds.

METHODS

Peripheral blood was collected from 25 dogs representing 13 pedigrees and seven crossbreeds. ATP-induced PS exposure on canine erythrocytes in vitro was assessed using a flow cytometric Annexin V binding assay.

RESULTS

ATP induced PS exposure in erythrocytes from all dogs studied. ATP caused PS exposure in a concentration-dependent manner with an EC50 value of 395 μmol/L. The non-P2X7 agonists, ADP or AMP, did not cause PS exposure. The P2X7 antagonist, AZ10606120, but not the P2X1 antagonist, NF449, blocked ATP-induced PS exposure.

CONCLUSION

The results indicate that ATP induces PS exposure in erythrocytes from various dog breeds and that this process is mediated by P2X7 activation.

Keywords: Adenosine triphosphate; Dog; P2X1 receptor; P2X7 receptor; Phospholipid; Purinergic receptor; Red blood cells

Core tip: Phosphatidylserine (PS) exposure in erythrocytes has potential roles in erythrocyte clearance and thrombus formation. Activation of the ATP-gated P2X7 receptor channel induces PS exposure in human erythrocytes, but whether this process occurs in erythrocytes from other mammals remained hitherto unknown. The current study shows that extracellular ATP causes PS exposure in dog erythrocytes from 13 pedigrees and seven crossbreeds. Notably, the current study shows that this process is mediated by P2X7 activation. These results suggest that P2X7-mediated PS exposure on erythrocytes may have important roles in red blood cell biology in dogs.