Molecular Physiological Analysis of Peptide Exocytosis and Pentidergic Neuromodulation.

肽胞吐作用和 Pentidergic 神经调节的分子生理学分析。

• 批准号: 12440237
• 负责人: OKA Yoshitaka
• 金额: $ 8.45万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12440237/
• 关键词: Peptide Neuron; Exocytosis; Neuromodulation; Molecular Physiology; Calcium Ion; Neurophysiology; Patch Clamp; GnRH; イメージング

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 slicesTo 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 … More e POA-GnRH slices.2. Exocytotic release of GnRHWe 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 GnRHTo 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

在本研究项目中，我们获得了以下有关促性腺激素释放激素（GnRH）系统的肽胞吐和肽能神经调节的结果。1.脑-垂体脑片释放GnRH的放射免疫测定（RIA）为了测定多个GnRH系统的释放活性，我们在各种条件下进行了脑-垂体脑片的静态孵育，并通过RIA测定释放到孵育液中的GnRH。我们证明高[K+]_0去极化刺激通过电压门控Ca^<2 +>通道的Ca ^<2 +>内流诱发GnRH释放。我们发现，在POA-GnRH切片中，GnRH释放量存在明显的性别差异。免疫组织化学分析使用的抗血清特异性的促性腺激素释放激素特异性POA-GnRH神经元显示存在的POA-GnRH神经元在男性比女性的数量大得多。这种明显的形态学性别差异被认为是性腺激素释放的基础。 ...更多信息 e POA-GnRH切片。GnRH的胞吐释放我们开发了一种使用碳纤维电极（CFE）实时电化学记录GnRH释放的方法。电流法测定GnRH的氧化电流。我们记录了硬骨鱼脑垂体切片垂体中的释放活动。CFE位于接受视前区GnRH神经元密集神经支配的垂体部分。在高[K^+]_0刺激下，以剂量依赖性方式记录到体电流。说明电流主要来源于GnRH的氧化电流，反映了GnRH的释放活性.为了研究TN-GnRH神经元活动的突触控制，我们在电生理学上分析了TN-GnRH神经元中谷氨酸受体（GluRs）的类型。通过使用各种特异性GluR激动剂和拮抗剂，我们发现它们具有离子型GluR（iGluR ; non-NMDAR和NMDAR）和第3组代谢型GluR（mGluR）。我们还发现了证据表明存在一种新型的iGluRs。少

项目成果

Tsutsui, H.: "Slow removal of Na^+ channel inactivation underlies the temporal filtering property in the teleost thalamic neurons"Journal of Physiology. 539. 743-753 (2002)

Tsutsui，H.：“Na + 通道失活的缓慢消除是硬骨鱼丘脑神经元中时间过滤特性的基础”生理学杂志。

Amano, M.: "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 (2002)

Amano, M.：“胸鳍鲽鱼大脑中黑色素浓缩激素的免疫细胞化学定位和个体发育”细胞和组织研究。

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)

Amano, M、Oka, Y.、Yamanome, T.、Okuzawa, K. 和 Yamamori, K.：“胸膜鲽鱼、条鲽鱼、Verasper moseri 的大脑和垂体中的三个 GnRH 系统”细胞和组织研究。

Kiya, T., and Oka, Y.: "Glutamate receptors in the terminal nerve gonadotropin-releasing hormone neurons of the dwarf gourami (teleost)"Neuroscience Letters. in press. (2003)

Kiya, T. 和 Oka, Y.：“侏儒古拉米（硬骨鱼）末端神经促性腺激素释放激素神经元中的谷氨酸受体”神经科学快报。

Amano, M., Oka, Y., Yamanome, T., Takahashi, A., 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 (20

Amano, M.、Oka, Y.、Yamanome, T.、Takahashi, A.、Yamamori, K.：“胸鳍鲽鱼大脑中黑色素浓缩激素的免疫细胞化学定位和个体发育”细胞和