A cholinergic model synapse to elucidate protein function at presynaptic terminals.
A large number of proteins have been identified at nerve terminals and a cascade of protein-protein interactions has been suggested to be involved in cycling of synaptic vesicle states. To explore protein function in presynaptic terminals, only a few unique synapses such as the squid giant synapse, the calyx of Held synapse and the hippocampal neuron autapse have been used. The squid giant synapse and the calyx of Held are useful to introduce reagents into their large presynaptic terminals and the hippocampal neuron autapse is a good system to modify a protein level by exogenous DNA or RNA. The cholinergic synapse formed between superior cervical ganglion (SCG) neurons in long-term culture is a useful model for a fast synapse. The axon of the large cell body contacts with soma of neighboring neurons. The architecture of synaptic connections makes it possible to introduce reagents into the presynaptic terminals by diffusion from a cell body within a short time. Introduction of exogenous cDNA or siRNA performed by microinjection into a SCG neuron allows us to modulate the level of the protein of interest or to express mutant proteins in the neuron. Here, we describe use of the model SCG neuronal synapse to elucidate function of presynaptic proteins in mediating synaptic transmission.
Ma
Huan
H
Department of Physiology, Tokyo Medical University, 1-1, Shinjuku-6-chome, Shinjuku-ku, Tokyo 160-8402, Japan.
Mochida
Sumiko
S
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation / Q000379:methods; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006982:Hypertonic Solutions / Q000494:pharmacology; D008845:Microinjections / Q000379:methods; D008954:Models, Biological; D009474:Neurons / Q000166:cytology*; D018408:Patch-Clamp Techniques; D017729:Presynaptic Terminals / Q000187:drug effects / Q000502:physiology*; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D017783:Superior Cervical Ganglion / Q000166:cytology; D013569:Synapses / Q000187:drug effects / Q000502:physiology*; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism; D050053:TRPM Cation Channels / Q000737:chemistry / Q000235:genetics; D013997:Time Factors
D018076:DNA, Complementary; D006982:Hypertonic Solutions; D034741:RNA, Small Interfering; D050825:Synaptosomal-Associated Protein 25; D050053:TRPM Cation Channels; D049452:Green Fluorescent Proteins; C486584:Trpm7 protein, rat; D000109:Acetylcholine