title: Ultrastructural localization of high-affinity choline transporter in the rat neuromuscular junction: enrichment on synaptic vesicles.
abstract: In cholinergic neurons, Na(+)- and Cl(-)-dependent, hemicholinium-3-sensitive, high-affinity choline uptake system is thought to be the rate-limiting step in acetylcholine (ACh) synthesis. The system is highly regulated by neuronal activity; the choline uptake is increased by a condition in which ACh release is favored. Here we analyzed the ultrastructural localization of the high-affinity choline transporter (CHT) in the rat neuromuscular junctions with two separate antibodies. The majority (>90%) of immunogold labeling of CHT was observed on synaptic vesicles rather than the presynaptic plasma membrane. Less than 5% of the gold-silver particles were associated with the plasma membrane, and more than 70% of such particles were localized within or in close vicinity to presynaptic active zones. Our morphological data support the recent hypothesis that trafficking of CHT from synaptic vesicles to the plasma membrane couples neuronal activity and choline uptake.
authors.affiliation: Department of Neurology, Tokyo Metropolitan Institute for Neuroscience, Tokyo 183-8526, Japan.
authors.forename: Kazuko
authors.initials: K
authors.lastname: Nakata
authors.forename: Takashi
authors.initials: T
authors.lastname: Okuda
authors.forename: Hidemi
authors.initials: H
authors.lastname: Misawa
mesh_terms: D000818:Animals; D001921:Brain / Q000648:ultrastructure; D002352:Carrier Proteins / Q000648:ultrastructure*; D005260:Female; D016253:Microscopy, Immunoelectron; D009046:Motor Neurons / Q000648:ultrastructure; D009419:Nerve Tissue Proteins / Q000648:ultrastructure*; D009469:Neuromuscular Junction / Q000648:ultrastructure*; D050480:Plasma Membrane Neurotransmitter Transport Proteins; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D027981:Symporters; D013572:Synaptic Vesicles / Q000648:ultrastructure*
chemical_list: D002352:Carrier Proteins; D009419:Nerve Tissue Proteins; D050480:Plasma Membrane Neurotransmitter Transport Proteins; D027981:Symporters; C075746:Slc6a8 protein, rat