Cellular Physiological analysis of peptidergic exocytotic events using primary culture system of isolated GnRH neurons
| Project/Area Number |
20770054
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Animal physiology/Animal behavior
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| Research Institution | The University of Tokyo |
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Principal Investigator |
ABE Hideki The University of Tokyo, 大学院・理学系研究科, 助教 (90396804)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| 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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| Keywords | GnRH / 開口放出 / 脳・神経 / 神経修飾 / ペプチド放出 / 神経ペプチド / 興奮-分泌連関 / 細胞培養 / ペプチド / イメージング / 遺伝子導入 / 生理学 / 神経科学 / 神経生物学 / 細胞・組織 / シグナル伝達 |
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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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Report
(4 results)
Research Products
(63 results)
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[Journal Article] Regular pacemaker activities of GnRH 2 neurons in comparison with spontaneous activities of GnRH1 and 3 neurons recorded from GFP-transgenic medaka2010
Author(s)
Kanda, S., Nishikawa, K., Karigo, T., Okubo, K., Isomae, S., Abe, H., Kobayashi, D., Oka, Y.
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Journal Title
Endocrinology 151
Pages: 695-701
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Peer Reviewed
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[Presentation] Diurnal regulation of GnRH1 neuronal activities and gonadotropin subunit transcription in a daily spawning fish, medaka2009
Author(s)
Karigo, T., Kanda, S., Aikawa, M., Abe, H., Okubo, K., Oka, Y.
Organizer
39th Annual Meetings, Society for Neuroscience
Place of Presentation
Chicago, USA
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[Presentation] Electrophysiological and anatomical characterization of three distinct GnRH neuronal populations in the GFP transgenic medaka2008
Author(s)
Kanda, S., Okubo, K., Isomae, S., Abe, H., Kobayashi, D., Takeda, H., Aida, K., Nagahama, Y., and OKA, Y.
Organizer
38th Annual Meetings, Society for Neuroscience
Place of Presentation
Washington DC, USA
Year and Date
2008-11-17
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[Presentation] Electrophysiological and anatomical characterization of three distinct GnRH neuronal populations in the GFP transgenic medaka2008
Author(s)
Kanda, S., Okubo, K., Isomae, S., Abe, H., Kobayashi, D., Takeda, H., Aida, K., Nagahama, Y., OKA, Y.
Organizer
38th Annual Meetings, Society for Neuroscience
Place of Presentation
WashingtonDC, USA
Related Report
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