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.

使用分散的垂体细胞的原发性培养物，在大鼠前垂体中检测到多激素细胞的群体。1。优化了双标记方法以检测垂体原发性培养中的多激素细胞。 LHβ和GH的mRNA和激素蛋白的共定位用作模型。关于灵敏度和特异性，与HRP聚合物或生物素结合的抗体的三明治标记方法可用于本研究。具有TSA强化的免疫荧光标记也是自动图像分析的良好方法，可以计算双重标记的单元格。2。在正常骑自行车和卵巢切除的雌性大鼠的垂体分散体中检测到产生LH和GH的垂体细胞。在卵巢切除术后1周，LH/GH双标记细胞的种群高于雌性大鼠。特别是，GH蛋白阳性细胞的LHβmRNA阳性比例显着不同，这表明GnRH可能与LH/GH细胞的出现有关。3。还检测到用于转录因子PIT-1和LHβmRNA或蛋白质的双阳性细胞。这些细胞的外观在垂体大鼠的垂体分散液中比偶然大鼠的外观更高。有趣的是，PIT-1/LH细胞对总垂体细胞的百分比低于GH/LH细胞的预期，这表明只有一部分LH/GH细胞为PIT-1阳性。这应该在进一步的研究中阐明。