title: Molecular mechanisms that control initiation and termination of physiological depolarization-evoked transmitter release.
abstract: Ca(2+) is essential for physiological depolarization-evoked synchronous neurotransmitter release. But, whether Ca(2+) influx or another factor controls release initiation is still under debate. The time course of ACh release is controlled by a presynaptic inhibitory G protein-coupled autoreceptor (GPCR), whose agonist-binding affinity is voltage-sensitive. However, the relevance of this property for release control is not known. To resolve this question, we used pertussis toxin (PTX), which uncouples GPCR from its G(i/o) and in turn reduces the affinity of GPCR toward its agonist. We show that PTX enhances ACh and glutamate release (in mice and crayfish, respectively) and, most importantly, alters the time course of release without affecting Ca(2+) currents. These effects are not mediated by G(beta)gamma because its microinjection into the presynaptic terminal did not alter the time course of release. Also, PTX reduces the association of the GPCR with the exocytotic machinery, and this association is restored by the addition of agonist. We offer the following mechanism for control of initiation and termination of physiological depolarization-evoked transmitter release. At rest, release is under tonic block achieved by the transmitter-bound high-affinity presynaptic GPCR interacting with the exocytotic machinery. Upon depolarization, the GPCR uncouples from its G protein and consequently shifts to a low-affinity state toward the transmitter. The transmitter dissociates, the unbound GPCR detaches from the exocytotic machinery, and the tonic block is alleviated. The free machinery, together with Ca(2+) that had already entered, initiates release. Release terminates when the reverse occurs upon repolarization.
authors.affiliation: Department of Neurobiology, Hebrew University, Jerusalem 91904, Israel.
authors.forename: Yonatan M
authors.initials: YM
authors.lastname: Kupchik
authors.forename: Grigory
authors.initials: G
authors.lastname: Rashkovan
authors.forename: Lily
authors.initials: L
authors.lastname: Ohana
authors.forename: Tal
authors.initials: T
authors.lastname: Keren-Raifman
authors.forename: Nathan
authors.initials: N
authors.lastname: Dascal
authors.forename: Hanna
authors.initials: H
authors.lastname: Parnas
authors.forename: Itzchak
authors.initials: I
authors.lastname: Parnas
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D044387:GTP-Binding Protein beta Subunits / Q000378:metabolism; D044388:GTP-Binding Protein gamma Subunits / Q000378:metabolism; D051379:Mice; D037342:Pertussis Toxin / Q000494:pharmacology; D011485:Protein Binding; D050765:Qa-SNARE Proteins / Q000378:metabolism; D043585:Receptor, Muscarinic M2 / Q000378:metabolism; D013569:Synapses / Q000187:drug effects / Q000378:metabolism*; D009435:Synaptic Transmission / Q000187:drug effects; D013997:Time Factors
chemical_list: C118981:G-protein Beta gamma; D044387:GTP-Binding Protein beta Subunits; D044388:GTP-Binding Protein gamma Subunits; D050765:Qa-SNARE Proteins; D043585:Receptor, Muscarinic M2; D037342:Pertussis Toxin; D000109:Acetylcholine; D002118:Calcium