title: Glutamate uptake occurs at an early stage of synaptic vesicle recycling.
abstract: Rapid membrane recycling in nerve terminals is required to maintain rapid synaptic transmission. Following the fusion of synaptic vesicles with synaptic plasma membranes, recycling can occur via clathrin-coated vesicles (CCVs) [1-3]. The fate of these vesicles is uncertain: they could simply uncoat and acquire other proteins from the cytosol to regenerate synaptic vesicles or they may fuse with endosomal structures from which synaptic vesicles could then bud. We have purified both CCVs and synaptic vesicles from rat brain, and measured the ability of these vesicle fractions to take up the excitatory neurotransmitter glutamic acid. We found that the normalized levels of glutamate uptake by the two types of vesicle were very similar. For each vesicle fraction, uptake required ATP and Cl- and could be fully inhibited by the specific vacuolar proton pump (v-ATPase) inhibitor concanamycin. We suggest that this ability to refill vesicles with neurotransmitter at the earliest intermediate on the recycling pathway - the CCV - may allow uncoated vesicles to immediately enter the releasable pool without sacrificing the quantal nature of neurotransmitter release.
authors.affiliation: Physiological Laboratory, University of Liverpool, Crown Street, Liverpool, L69 3BX, UK.
authors.forename: I A
authors.initials: IA
authors.lastname: Prior
authors.forename: M J
authors.initials: MJ
authors.lastname: Clague
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001692:Biological Transport; D001921:Brain / Q000502:physiology; D002458:Cell Fractionation; D002712:Chlorides / Q000378:metabolism; D003600:Cytosol / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018698:Glutamic Acid / Q000378:metabolism*; D018942:Macrolides; D008561:Membrane Fusion; D009411:Nerve Endings / Q000502:physiology; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors; D051381:Rats; D013570:Synaptic Membranes / Q000502:physiology* / Q000648:ultrastructure; D013572:Synaptic Vesicles / Q000502:physiology* / Q000648:ultrastructure; D025262:Vacuolar Proton-Translocating ATPases
chemical_list: D000900:Anti-Bacterial Agents; D002712:Chlorides; D004791:Enzyme Inhibitors; D018942:Macrolides; D018698:Glutamic Acid; C036978:concanamycin A; D000255:Adenosine Triphosphate; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases