title: P2X purinergic receptor knockout mice reveal endogenous ATP modulation of both vasopressin and oxytocin release from the intact neurohypophysis.
abstract: Bursts of action potentials are crucial for neuropeptide release from the hypothalamic neurohypophysial system (HNS). The biophysical properties of the ion channels involved in the release of these neuropeptides, however, cannot explain the efficacy of such bursting patterns on secretion. We have previously shown that ATP, acting via P2X receptors, potentiates only vasopressin (AVP) release from HNS terminals, whereas its metabolite adenosine, via A1 receptors acting on transient Ca(2+) currents, inhibits both AVP and oxytocin (OT) secretion. Thus, purinergic feedback-mechanisms have been proposed to explain bursting efficacy at HNS terminals. Therefore, in the present study, we have used specific P2X receptor knockout (rKO) mice and purportedly selective P2X receptor antagonists to determine the P2X receptor subtype responsible for endogenous ATP induced potentiation of electrically-stimulated neuropeptide release. Intact neurohypophyses (NH) from wild-type (WT), P2X3 rKO, P2X2/3 rKO and P2X7 rKO mice were electrically stimulated with four 25-s bursts (3 V at 39 Hz) separated by 21-s interburst intervals with or without the P2X2 and P2X3 receptor antagonists, suramin or pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS). These frequencies, number of bursts, and voltages were determined to maximise both AVP and OT release by electrical stimulations. Treatment of WT mouse NH with suramin/PPADS significantly reduced electrically-stimulated AVP release. A similar inhibition by suramin was observed in electrically-stimulated NH from P2X3 and P2X7 rKO mice but not P2X2/3 rKO mice, indicating that endogenous ATP facilitation of electrically-stimulated AVP release is mediated primarily by the activation of the P2X2 receptor. Unexpectedly, electrically-stimulated OT release from WT, P2X3, P2X2/3 and P2X7 rKO mice was potentiated by suramin, indicating nonpurinergic effects by this 'selective' antagonist. Nevertheless, these results show that sufficient endogenous ATP is released by bursts of action potentials to act at P2X2 receptors in a positive-feedback mechanism to 'differentially' modulate neuropeptide release from central nervous system terminals.
authors.affiliation: Department of Microbiology and Physiological Systems & Program in Neuroscience, University of Massachusetts Medical School, Worcester, MA 01655, USA.
authors.forename: E E
authors.initials: EE
authors.lastname: Custer
authors.forename: T K
authors.initials: TK
authors.lastname: Knott
authors.forename: A E
authors.initials: AE
authors.lastname: Cuadra
authors.forename: S
authors.initials: S
authors.lastname: Ortiz-Miranda
authors.forename: J R
authors.initials: JR
authors.lastname: Lemos
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000255:Adenosine Triphosphate / Q000502:physiology*; D000818:Animals; D001127:Arginine Vasopressin / Q000378:metabolism*; D004558:Electric Stimulation / Q000379:methods; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008954:Models, Biological; D010121:Oxytocin / Q000378:metabolism*; D010904:Pituitary Gland, Posterior / Q000187:drug effects / Q000378:metabolism*; D058920:Purinergic P2X Receptor Antagonists / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D058476:Receptors, Purinergic P2X2 / Q000235:genetics / Q000502:physiology*; D058477:Receptors, Purinergic P2X3 / Q000235:genetics / Q000502:physiology*; D013498:Suramin / Q000494:pharmacology
chemical_list: D058920:Purinergic P2X Receptor Antagonists; D058476:Receptors, Purinergic P2X2; D058477:Receptors, Purinergic P2X3; D001127:Arginine Vasopressin; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; D010121:Oxytocin; D011732:Pyridoxal Phosphate; D013498:Suramin; D000255:Adenosine Triphosphate