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.

采用分散垂体细胞原代培养法检测大鼠垂体前叶多激素细胞的数量。优化双标记法检测垂体原代培养的多激素细胞。以LH β和GH mRNA和激素蛋白共定位为模型。从敏感性和特异性两方面考虑，本研究可采用HRP聚合物偶联抗体或生物素夹心标记法。免疫荧光标记与TSA增强也是自动图像分析计数双标记细胞的好方法。在正常周期和去卵巢的雌性大鼠垂体弥散体中检测到产生LH和GH的垂体细胞。去卵巢1周后大鼠的LH/GH双标记细胞数量明显高于发情大鼠。特别是GH蛋白阳性细胞的LH β mrna阳性比例有显著差异，提示GnRH可能与LH/GH细胞的出现有关。同时检测到转录因子Pit-1和LH β mRNA或蛋白双阳性细胞。发情大鼠垂体弥散中这些细胞出现的比例高于发情大鼠。有趣的是，垂体总细胞中Pit-1/LH细胞的比例低于GH/LH细胞的比例，这表明只有一部分LH/GH细胞是Pit-1阳性。这一点应在进一步的研究中加以澄清。