Population of Multi-hormonal cells in the anterior pituitary gland

垂体前叶多激素细胞群

• 批准号: 17590175
• 负责人: KUDO Akihiko
• 金额: $ 2.43万
• 依托单位: Kyorin University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17590175/
• 关键词: tissues and cells; hormone; pituitary gland; gene expression; cell differentiation; international communication; U. S. A.; 細胞・組織

Population of multi-hormonal cells were detected in rat anterior pituitary gland, using primary culture of dispersed pituitary cells.1. The double-labeling method was optimized to detect multi-hormonal cells in pituitary primary culture. Colocalization of mRNA and hormone protein of LH beta and GH was used as a model. Regarding to both sensitivity and specificity, sandwich labeling method with antibodies conjugated with HRP polymer or with biotin can be used to this study. Immunofluorescent labeling with TSA intensification was also good method for automated image analysis to count double labeled cells.2. Pituitary cells producing LH and GH were detected in pituitary dispersion from normal cycling and ovariectomized female rats. In the rats 1 week after ovariectomized, the population of LH/GH double-labeled cells was higher than that of proestrous rats. Particularly, LH beta mRNA-positive ratio of GH protein-positive cells was significantly different It is suggested that GnRH may be concerned with the appearance of LH/GH cells.3. Double-positive cells for transcription factor Pit-1 and LH beta mRNA or protein were also detected. The ratio of appearance of those cells was higher in pituitary dispersion from proestrous rats than from diestrous rats. Interestingly, the percentage of Pit-1/LH cells to total pituitary cells was lower than expected by that of GH/LH cells, suggesting only a portion of LH/GH cells is Pit-1 positive. This should be clarified in the further study.

采用分散的垂体细胞原代培养方法，在大鼠脑垂体前叶检测到多激素细胞群。对双标记法进行了优化，以检测原代培养的垂体多激素细胞。以促黄体生成素β和生长激素的信使核糖核酸和激素蛋白共定位为模型。就敏感性和特异性而言，抗体与HRP聚合物或生物素结合的夹心标记法均可用于本研究。结论：1.免疫荧光标记结合TSA增强也是自动图像分析计数双标记细胞的良好方法。正常周期雌性大鼠和去卵巢雌性大鼠的脑垂体液中可检测到分泌黄体生成素和生长激素的细胞。卵巢摘除后1周，黄体生成素/生长激素双标细胞数量高于发情前期大鼠。尤其是GH蛋白阳性细胞间的LHβmRNA阳性率有显著差异，提示GnRH可能与LHGH细胞的出现有关。同时检测到转录因子Pit-1和黄体生成素β基因或蛋白的双阳性细胞。这些细胞在发情前期大鼠脑垂体液中的出现率高于发情间期大鼠。有趣的是，Pit-1/Lh细胞占总垂体腺细胞的百分比低于GH/Lh细胞，表明只有部分Lh/GH细胞呈Pit-1阳性。这一点应该在进一步的研究中得到澄清。