Dual presynaptic control by ATP of glutamate release via facilitatory P2X1, P2X2/3, and P2X3 and inhibitory P2Y1, P2Y2, and/or P2Y4 receptors in the rat hippocampus.
ATP is released in a vesicular manner from nerve terminals mainly at higher stimulation frequencies. There is a robust expression of ATP (P2) receptors in the brain, but their role is primarily unknown. We report that ATP analogs biphasically modulate the evoked release of glutamate from purified nerve terminals of the rat hippocampus, the facilitation being mediated by P2X1, P2X2/3, and P2X3 [antagonized by 8-(benzamido)naphthalene-1,3,5-trisulfonate and 2',3'-O-(2,4,6-trinitrophenyl)-ATP] and the inhibition by P2Y1, P2Y2, and/or P2Y4 [antagonized by reactive blue 2 and 2'deoxy-N6-methyladenosine-3',5'-bisphosphate and mimicked by P1-(urinine 5'-),P4-(inosine 5'-) tetraphosphate and 2-methylthio-ADP] receptors. The combination of single-cell PCR analysis of rat hippocampal pyramidal neurons, Western blot analysis of purified presynaptic active zone fraction, and immunocytochemical analysis of hippocampal glutamatergic terminals revealed that the P2 receptors expressed in glutamatergic neurons, located in the active zone and in glutamatergic terminals, were precisely P2X1, P2X2, and P2X3 subunits and P2Y1, P2Y2, and P2Y4 receptors. This provides coincident functional and molecular evidence that P2 receptors are present and act presynaptically as a modulatory system controlling hippocampal glutamate release.
Centre for Neurosciences of Coimbra, Institute of Biochemistry, Faculty of Medicine, University of Coimbra, 3004-504 Coimbra, Portugal.
Ricardo J
RJ
Rodrigues
Teresa
T
Almeida
Peter J
PJ
Richardson
Catarina R
CR
Oliveira
Rodrigo A
RA
Cunha
D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology / Q000502:physiology*; D000266:Adenylyl Imidodiphosphate / Q000494:pharmacology; D000818:Animals; D001254:Astrocytoma / Q000378:metabolism / Q000473:pathology; D002118:Calcium / Q000032:analysis; D002460:Cell Line / Q000378:metabolism; D045744:Cell Line, Tumor / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism*; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D007668:Kidney; D008297:Male; D011188:Potassium / Q000494:pharmacology; D017966:Pyramidal Cells / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D017661:Receptors, Presynaptic / Q000187:drug effects / Q000502:physiology; D018048:Receptors, Purinergic P2 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D058469:Receptors, Purinergic P2X; D058476:Receptors, Purinergic P2X2; D058477:Receptors, Purinergic P2X3; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013347:Subcellular Fractions / Q000378:metabolism; D013498:Suramin / Q000031:analogs & derivatives / Q000494:pharmacology; D013574:Synaptosomes / Q000737:chemistry / Q000378:metabolism; D014162:Transfection; D014227:Triazines / Q000494:pharmacology; D014230:Triazoles / Q000494:pharmacology; D014970:Xanthines / Q000494:pharmacology
C111914:N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate; C105374:NF023; C546403:P2rx2 protein, rat; C546410:P2rx3 protein, rat; C546384:P2ry1 protein, rat; C546387:P2ry2 protein, rat; D012333:RNA, Messenger; D017661:Receptors, Presynaptic; D018048:Receptors, Purinergic P2; D058469:Receptors, Purinergic P2X; D058476:Receptors, Purinergic P2X2; D058477:Receptors, Purinergic P2X3; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D011993:Recombinant Fusion Proteins; D014227:Triazines; D014230:Triazoles; D014970:Xanthines; C097270:ZM 241385; C104749:purinoceptor P2Y4; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; D000266:Adenylyl Imidodiphosphate; D018698:Glutamic Acid; C012842:Cibacron Blue F 3GA; D011732:Pyridoxal Phosphate; D013498:Suramin; C045674:2',3'-O-(2,4,6-trinitro-cyclohexadienylidine)adenosine 5'-triphosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C051360:1,3-dipropyl-8-cyclopentylxanthine; C002630:alpha,beta-methyleneadenosine 5'-triphosphate; D011188:Potassium; D002118:Calcium