The mechanism of neurotransmitter release and its control using electric organ synapse of electric ray

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1. cDNA cloning of voltage-dependent Ca channel of electromotor neuron from electric ray.RT-PCR was carried out with electric lobe of Japanese electric ray, Narke Japonica. Partial sequence of N- and L-type voltage dependent Ca channel were obtained.2. Experimental setup to investigate the electrophysiological characteristics of Ca channel of electric organ nerve terminal.In order to do the electrophysiological characterization of Ca channel of electric organ synaptosomes, lipid bilayer system was introduced.Preliminary experiments revealed that a voltage dependent anion channel was detected in ciliary membrane vesicles from Teterahymena.3. Asymmetrical distribution of phospholipids in membrane of synaptosome and synaptic vesicle from electric ray.The asymmetrical distribution of phospholipids between the outer and inner leaflets of synaptosome and synaptic vesicle was investigated. In both membranes, phosphatidylserine (PS) was dominant in the leaflet which faces cytoplasm.4. The effects of adenosine on the acetylcholine release from electric organ synaptosomes.The effects of adenosine on the acetylcholine (ACh) release from synaptosomes was investigated. Adenosine inhibited the ACh release through adenosine Al receptor and facilitated the release through A2 receptor. The inhibitory effect via Al receptor was caused by the inhibition of N-type voltage dependent Ca channel, and the facilitatory effects via A2 receptor was caused by the activation of P-type Ca channel.