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The P2X7 receptor mediates the uptake of organic cations in canine erythrocytes and mononuclear leukocytes: comparison to equivalent human cell types

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Abstract

We previously demonstrated that canine erythrocytes express the P2X7 receptor, and that the function and expression of this receptor is greatly increased compared with human erythrocytes. Using 86Rb+ (K+) and organic cation flux measurements, we further compared P2X7 in erythrocytes and mononuclear leukocytes from these species. Concentration response curves of BzATP- and ATP-induced 86Rb+ efflux demonstrated that canine P2X7 was less sensitive to inhibition by extracellular Na+ ions compared to human P2X7. In contrast, canine and human P2X7 showed a similar sensitivity to the P2X7 antagonists KN-62 and Mg2+. KN-62 and Mg2+ also inhibited ATP-induced choline+ uptake into canine and human erythrocytes. BzATP and ATP but not ADP or NAD induced ethidium+ uptake into canine monocytes, T- and B-cells. ATP-induced ethidium+ uptake was twofold greater in canine T-cells compared to canine B-cells and monocytes. KN-62 inhibited the ATP-induced ethidium+ uptake in each cell type. P2X7-mediated uptake of organic cations was 40- and fivefold greater in canine erythrocytes and lymphocytes (T- and B-cells), respectively, compared to equivalent human cell types. In contrast, P2X7 function was threefold lower in canine monocytes compared to human monocytes. Thus, P2X7 activation can induce the uptake of organic cations into canine erythrocytes and mononuclear leukocytes, but the relative levels of P2X7 function differ to that of equivalent human cell types.

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Abbreviations

BSA:

bovine serum albumin

ADP:

adenosine 5′-diphosphate

ATP:

adenosine 5′-triphosphate

BzATP:

2′- and 3′-0(4-benzoylbenzoyl) ATP

DMSO:

dimethyl sulphoxide

FITC:

fluorescein isothiocyanate

mAb:

monoclonal antibody

NAD:

β-nicotinamide adenine dinucleotide

SEM:

standard error of the mean

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Authors and Affiliations

  1. Department of Medicine, Nepean Clinical School, University of Sydney, Penrith, NSW, Australia

    Ryan O. Stevenson, James S. Wiley & Ronald Sluyter

  2. Faculty of Veterinary Science, University of Sydney, Sydney, NSW, Australia

    Ryan O. Stevenson & Rosanne M. Taylor

  3. School of Biological Sciences, University of Wollongong, Wollongong, NSW, Australia

    Ronald Sluyter

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  1. Ryan O. Stevenson
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  2. Rosanne M. Taylor
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  3. James S. Wiley
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Correspondence to Ronald Sluyter.

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Stevenson, R.O., Taylor, R.M., Wiley, J.S. et al. The P2X7 receptor mediates the uptake of organic cations in canine erythrocytes and mononuclear leukocytes: comparison to equivalent human cell types. Purinergic Signalling 5, 385–394 (2009). https://doi.org/10.1007/s11302-009-9163-1

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