| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2010: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2009: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2008: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
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| Research Abstract |
To understand the excitation-secretion dynamics of peptidergic neuron from various cellular compartments, we recently established a primary culture system of isolated fish terminal nerve (TN)-gonadotropin releasing hormone (GnRH) neuron. In this study, we examined the electrophysiological properties of the cultured TN-GnRH neurons. They showed subthreshold spontaneous membrane potential oscillations and could be induced to fire in phasic or tonic patterns. Then, we applied a single cell electroporation method for introducing exogenous genes to visually identified TN-GnRH neurons. We successfully introduced plasmids encoding fusion proteins of GFP and exocytotic vesicle components to identified TN-GnRH neurons. Exogenous expression of those genes constructs showed distributions of punctate fluorescence in the soma and neurites. Time-lapse imaging of those fluorescence enabled visualization of movements and exocytosis of peptides in a peptidergic neuron.
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