[(35)S]GTPgammaS autoradiography reveals a wide distribution of G(i/o)-linked ADP receptors in the nervous system: close similarities with the platelet P2Y(ADP) receptor.
No G(i)-linked P2Y receptors have been cloned to date but the presence of such receptors is thought to be restricted to platelets and certain clonal cell lines. Using the functional approach of [(35)S]guanosine 5'-[gamma-thio]-triphosphate autoradiography, we uncovered the widespread presence of such receptors in the CNS. Under conditions in which the prominent signal due to tonic adenosine receptor activity is masked, ADP and ATP stimulated G-protein activity in multiple grey and white matter regions. Localization in the grey matter suggests inhibitory auto-/heteroreceptor function. In the white matter, activated G proteins appeared as 'hot spots' (presumed oligodendrocyte progenitors) with scattered distribution along the main fibre tracts. Responses to ATP were diminished under conditions that inhibited degradation, suggesting that prior conversion to ADP explained agonist action. Uracil nucleotides were ineffective but 2-methylthio-ADP activated G proteins approximately 500-fold more potently than ADP, although both were similarly degraded. Throughout the brain, ADP-dependent G-protein activity was reversed by 2-hexylthio-AdoOC(O)Asp(2), a non-phosphate ATP analogue, whereas selective P2Y(1) receptor antagonists proved ineffective. A similar receptor was also disclosed from the adrenal medulla. These data witness a hitherto unrecognized abundance of G(i/o)-linked ADP receptors in the nervous system. Biochemical and pharmacological behaviour suggests striking similarities to the elusive platelet P2Y(ADP) receptor.
Laitinen
J T
JT
Department of Physiology, University of Kuopio, Finland Institute of Chemical Physics, Tartu University, Estonia Department of Neuroscience and Neurology, University and University Hospital of Kuopio, Finland. Jarmo.Laitinen@uku.fi
Uri
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Raidaru
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Miettinen
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D000227:Adenine Nucleotides / Q000494:pharmacology; D000241:Adenosine / Q000031:analogs & derivatives / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000313:Adrenal Medulla / Q000737:chemistry; D000818:Animals; D001345:Autoradiography; D001792:Blood Platelets / Q000737:chemistry*; D001921:Brain / Q000098:blood supply; D001923:Brain Chemistry; D002196:Capillaries / Q000166:cytology; D002460:Cell Line; D004229:Dithiothreitol / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000737:chemistry; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000032:analysis*; D005910:Glioma / Q000473:pathology; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000032:analysis* / Q000378:metabolism; D020962:Heterotrimeric GTP-Binding Proteins / Q000032:analysis*; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D009457:Neuroglia / Q000737:chemistry; D009928:Organ Specificity; D051381:Rats; D017208:Rats, Wistar; D018048:Receptors, Purinergic P2 / Q000032:analysis* / Q000737:chemistry* / Q000187:drug effects; D058925:Receptors, Purinergic P2Y12; D015398:Signal Transduction; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013462:Sulfur Radioisotopes / Q000032:analysis; D013873:Thionucleotides / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014500:Uracil Nucleotides / Q000494:pharmacology; D014970:Xanthines / Q000378:metabolism
C425778:2-hexylthio-AdoOC(O)Asp2; C092431:6-N,N-diethyl-beta,gamma-dibromomethylene-D-ATP; D000227:Adenine Nucleotides; D004791:Enzyme Inhibitors; D046911:Macromolecular Substances; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C553058:P2ry12 protein, rat; D018048:Receptors, Purinergic P2; D058925:Receptors, Purinergic P2Y12; D013439:Sulfhydryl Reagents; D013462:Sulfur Radioisotopes; D013873:Thionucleotides; D014500:Uracil Nucleotides; D014970:Xanthines; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C051360:1,3-dipropyl-8-cyclopentylxanthine; D000251:Adenosine Triphosphatases; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D020962:Heterotrimeric GTP-Binding Proteins; D000241:Adenosine; D004229:Dithiothreitol; C008501:2-methylthio-ATP