Analysis of neuronal circuit for the gonadotropin-releasing hormone surege generator (GnRH) using an in vitro GnRH surge generator model

使用体外 GnRH 浪涌发生器模型分析促性腺激素释放激素 Surege 发生器 (GnRH) 的神经元回路

• 批准号: 11680790
• 负责人: FUNABASHI Toshiya
• 金额: $ 0.51万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680790/
• 关键词: Rat; GnRH surge generator; Vasopressin; co-culture; Preoptic area; GABA neurons; acute slice; Suprachiasmatic nucleus; エストロジェン; LHサージ; バソプレッシン; 培養; 視索前野; 視交叉上核; カルシウムイメージング; イメージング; サージジェネレーター; GnRH

1. We determined morphological changes in an in vitro gonadotropin-releasing hormone (GnRH) surge generator model we developed. GnRH neurons in the POA slice were clearly visible 4 weeks after co-culture with the suprachiasmatic nucleus (SCN). However, we could not stained vasopressin neurons. We tried to identify living GnRH neurons by transfecting a expression vector containing GnRH promoter and Ds-Red as a reporter. Transfection efficiency of this vector is, however, very low even in GT1-7 cell lines.2. We investigated the propagation of electrical stimulus-evoked excitation in the in vitro GnRH surge generator model. We found that the evoked excitation was not propagated to the POA slice when an electrical stimulus was applied in the SCN slice, suggesting that the POA slice could not make synaptic contact with the SCN slice.3. We next measured GnRH releases in tne POA slice obtained from the adult rat on the day of proestrus. Bicuculline, an GABA_A receptor antagonist, significantly increased GnRH releases in the POA slice. Interestingly, when the POA slice was perfused with the SCN slice, we found that GnRH release was significantly fractuated, suggesting that GnRH release was conveyed by the SCN via a humoral information.From these results, we conclude that an acute slice preparation is useful tools to investigate the GnRH surge generator activity.

1.我们确定了我们开发的体外促性腺激素释放激素（GnRH）浪涌发生器模型的形态学变化。与视交叉上核（SCN）共培养4周后，POA切片中的GnRH神经元清晰可见。然而，我们不能染色加压素神经元。我们尝试通过构建含有GnRH启动子和Ds-Red报告基因的表达载体来鉴定活的GnRH神经元。然而，即使在GT 1 -7细胞系中，该载体的转染效率也很低.我们研究了电刺激诱发的兴奋在体外GnRH浪涌发生器模型的传播。我们发现，当电刺激施加于SCN脑片时，诱发兴奋并没有传播到POA脑片，提示POA脑片不能与SCN脑片形成突触接触.我们接下来测量了发情前期大鼠POA切片中GnRH的释放。GABA_A受体拮抗剂荷包牡丹碱能显著增加POA脑片的GnRH释放。有趣的是，当POA切片与SCN切片灌注时，我们发现GnRH释放显著减少，这表明GnRH释放是通过SCN通过体液信息传递的。

项目成果

Funabashi T, Aiba S, Sano A, Shinohara K, Kimura F: "Intracerebroventricular injection of arginine-vasopressin V1 receptor antagonist attenuates the surge of luteinizing hormone and prolactin secretion in proestrous rats"Neuroscience Letters. 260. 37-40 (

Funabashi T、Aiba S、Sano A、Shinohara K、Kimura F：“脑室内注射精氨酸-加压素 V1 受体拮抗剂可减弱发情前大鼠黄体生成素和催乳素分泌的激增”《神经科学快报》。

Funabashi T, Shinohara K, Mitsushima D and Kimura F: "Gonadotropin-releasing horm one exhibits circadian rhythm in phase with arginine-vasopressin in co-cultures of the female rat preoptic area and suprachiasmatic nucleus"J Neuroendocrinol. (in press). (2

Funabashi T、Shinohara K、Mitsushima D 和 Kimura F：“在雌性大鼠视前区和视交叉上核的共培养物中，促性腺激素释放激素 1 表现出与精氨酸-加压素同相的昼夜节律”J Neuroendocrinol。

Funabashi T, Shinohara K, Kimura F: "Neuronal control circuit for the gonadotropin-releasing hormone surge in rats. In / Neuroplasticity, development, and steroid hormone action"Handa RJ, Hayashi S, Terasawa E, Kawata M(des) CRC Press, Florida. 405 (2002)

Funabashi T、Shinohara K、Kimura F：“大鼠促性腺激素释放激素激增的神经控制回路。/神经可塑性、发育和类固醇激素作用”Handa RJ、Hayashi S、Terasawa E、Kawata M(des) CRC Press

舩橋利也, 貴邑冨久子: "内側視索前野GnRHニューロンと性周期"Clinical Neuroscience. 19. 1122-1124 (2001)

Toshiya Funahashi、Tomiko Takamura：“内侧视前区 GnRH 神经元和性周期”《临床神经科学》19. 1122-1124 (2001)。

Funabashi T, Mitsushima D, Nakamura IT, Shinohara K, Suyama K, Kimura F: "Gonadotropin-releasing hormone (GnRH) surge generator in female rats"Progress in Brain Research (special issue for 2nd international symposium on the comparative biology of GnRH). (

Funabashi T、Mitsushima D、Nakamura IT、Shinohara K、Suyama K、Kimura F：“雌性大鼠促性腺激素释放激素（GnRH）激增发生器”脑研究进展（第二届 GnRH 比较生物学国际研讨会特刊）