Activity-dependent autocrine-paracrine activation of neuronal P2Y receptors.
Activation of P2Y receptors by released nucleotides subserves important autocrine-paracrine functions in various non-neural tissues. To investigate how P2Y receptors are activated in a neuronal environment, we used PC12 cells in which nucleotides were found to elicit increases in inositol phosphates via P2Y2 and decreases in cAMP via P2Y12 receptors. Depolarization of PC12 cells raised inositol phosphates, and blockade of voltage-gated Ca2+ channels by Cd2+ or degradation of extracellular nucleotides by apyrase prevented this effect. In nondepolarized cells, apyrase did not affect inositol phosphates. Depolarization of PC12 cells also reduced the A2A receptor-mediated synthesis of cAMP. This effect was again prevented by Cd2+ or apyrase, but apyrase enhanced the synthesis of cAMP even in nondepolarized cells. Overexpression of rat P2Y2 receptors increased the nucleotide-dependent inositol phosphate accumulation and enhanced the effect of K+ depolarization. Nevertheless, apyrase still failed to alter spontaneous inositol phosphate accumulation. Expression of rat P2Y1 receptors, in contrast, led to huge increases in spontaneous inositol phosphate accumulation, which was reduced by a receptor antagonist or by apyrase. This increased synthesis of inositol phosphates could not be further enhanced by depolarization or receptor agonists, but when endogenous nucleotides were removed by superfusion, recombinant P2Y1 receptors could be activated to mediate an inhibition of M-type K+ channels. These results indicate that nucleoside diphosphate-sensitive (P2Y12 and P2Y1) receptors are activated by spontaneous nucleotide release, whereas triphosphate-sensitive (P2Y2) receptors require an excess of depolarization-evoked release to become activated.
Moskvina
Eugenia
E
Institute of Pharmacology, University of Vienna, A-1090 Vienna, Austria.
Unterberger
Ursula
U
Boehm
Stefan
S
D000818:Animals; D019898:Autocrine Communication; D000242:Cyclic AMP / Q000378:metabolism; D004553:Electric Conductivity; D007295:Inositol Phosphates / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D009711:Nucleotides / Q000494:pharmacology; D016716:PC12 Cells; D019899:Paracrine Communication; D018408:Patch-Clamp Techniques; D058910:Purinergic P2 Receptor Agonists; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2
D007295:Inositol Phosphates; D009711:Nucleotides; C546384:P2ry1 protein, rat; C546387:P2ry2 protein, rat; D058910:Purinergic P2 Receptor Agonists; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D058473:Receptors, Purinergic P2Y2; D000242:Cyclic AMP