2002 Fiscal Year Final Research Report Summary
Molecular Physiological Analysis of Peptide Exocytosis and Pentidergic Neuromodulation.
| Project/Area Number |
12440237
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
動物生理・代謝
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| Research Institution | The University of Tokyo |
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Principal Investigator |
OKA Yoshitaka The University of Tokyo, Graduate School of Science, Associate Professor, 大学院・理学系研究科, 助教授 (70143360)
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| Co-Investigator(Kenkyū-buntansha) |
TOGO Tatsuru The University of Tokyo, Graduate School of Science, Assistant professor, 大学院・理学系研究科, 助手 (40334247)
YOSHIDA Manabu The University of Tokyo, Graduate School of Science, Assistant professor, 大学院・理学系研究科, 助手 (60301785)
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| Project Period (FY) |
2000 – 2002
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| Keywords | Peptide Neuron / Exocytosis / Neuromodulation / Molecular Physiology / Calcium Ion / Neurophysiology / Patch Clamp / GnRH |
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| Research Abstract |
In the present research project, we obtained the following results concerning the peptide exocytosis and peptidergic neuromodulation of the gonadotropin-releasing hormone (GnRH) system..
1. Radioimmunoassay (RIA) of GnRH release from brain-pituitary slices
To measure GnRH release activities from multiple GnRH systems, we conducted a static incubation of brain- pituitary slices under various conditions, and GnRH released into the incubation medium was measured by RIA. We demonstrated that GnRH release was evoked by high [K+]_0 depolarizing stimuli via Ca^<2+> influx through voltage-gated Ca^<2+> channels. We revealed the presence of conspicuous sexual difference in the amount of GnRH release in the POA-GnRH slices. Immunohistochemical analysis using an antiserum specific to the GnRH specific to POA-GnRH neurons revealed presence of much larger number of POA-GnRH neurons in males than in females. This clear morphological sexual difference is suggested to underlie that of GnRH release in th
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e POA-GnRH slices.
2. Exocytotic release of GnRH
We developed a method for real-time electrochemical recording of GnRH release using carbon fiber electrodes (CFE). The oxidation current of GnRH was measured by amperometry. We recorded the release activity in the pituitary of the teleost brain-pituitary slice. The CFE was located in the part of the pituitary that receives dense innervation from GnRH neurons in the preoptic area. A bulk amperometric current was recorded in response to high [K^+]_0 stimulation in a dose dependent manner. We suggested that the amperometric currents are mainly attributed to the oxidation currents of GnRH and reflect the GnRH release activity.
3. Glutamatergic synaptic inputs to GnRH
To investigate the synaptic control of activities of the TN-GnRH neurons, we analyzed electrophysiologically the type of glutamate receptors (GluRs) in the TN-GnRH neurons. By using various specific GluR agonists and antagonists, we found that they have ionotropic GluRs (iGluR ; non-NMDAR and NMDAR) and group 3 metabotropic GluRs (mGluR). We also found evidence to suggest the presence of a novel type of iGluRs. Less
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[Publications] Kim, M., Kobayashi, M., Oka, Y., Amano, M., Kawashima, S., and Aida, K.: "Effects of olfactory tract section on the immunohistochemical distribution of brain GnRH in the female goldfish, Carassius auratus"Zoological Science. 18. 241-248 (2001)
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[Publications] Amano, M, Oka, Y., Yamanome, T., Okuzawa, K., and Yamamori, K.: "Three GnRH systems in the brain and pituitary of a pleuronectiform fish, barfin flounder, Verasper moseri"Cell and Tissue Research. 309. 323-329 (2002)
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[Publications] Amano, M., Oka, Y., Yamanome, T., Takahashi, A.,and Yamamori, K.: "Immunocytochemical localization and ontogenic development of melanin-concentrating hormone in the brain of a pleuronectiform fish, the barfin flounder"Cell and Tissue Research. 311. 71-77 (2003)
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[Publications] Morisawa, M., Izumi, H., Yoshida, M., and Oka, Y. , eds. by H. Sawada,H. Yokosawa, and C.C. Lambert: "Cell signaling for activation of motility and chemotaxis in the sperm of Ciona In : "The Biology of Ascidians""Springer Verlag. 86-91 (2001)
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