2010 Fiscal Year Final Research Report
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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| 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 |
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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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[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
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
Year and Date
20091000
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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
Year and Date
20081100
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