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群。 1．优化双标记方法以检测垂体原代培养物中的多激素细胞。 LH β 和 GH 的 mRNA 和激素蛋白的共定位被用作模型。就敏感性和特异性而言，本研究可采用与 HRP 聚合物或生物素缀合的抗体夹心标记方法。 TSA增强免疫荧光标记也是自动图像分析计数双标记细胞的好方法。2．在正常周期和卵巢切除雌性大鼠的垂体分散体中检测到产生 LH 和 GH 的垂体细胞。去势大鼠1周后LH/GH双标细胞数量较发情前期大鼠有所增加。特别是GH蛋白阳性细胞中LHβmRNA阳性率差异显着，提示GnRH可能与LH/GH细胞的出现有关。 3．还检测到转录因子 Pit-1 和 LH beta mRNA 或蛋白的双阳性细胞。这些细胞在发情前期大鼠的垂体弥散中出现的比例高于发情期大鼠的垂体弥散中。有趣的是，Pit-1/LH 细胞占垂体细胞总数的百分比低于 GH/LH 细胞的预期，表明只有一部分 LH/GH 细胞呈 Pit-1 阳性。这应该在进一步的研究中得到澄清。