GFP-expressing S100β-positive cells of the rat anterior pituitary differentiate into hormone-producing cells
GFP-expressing S100β-positive cells of the rat anterior pituitary differentiate into hormone-producing cells
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DOI：
10.1007/s00441-014-1890-0
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发表时间：
2014-09-01
影响因子：
3.6
通讯作者：
Kato, Yukio
中科院分区：
文献类型：
--
作者：
Higuchi, Masashi;Kanno, Naoko;Kato, Yukio
垂体前叶中的一些非内分泌细胞负责提供干细胞/祖细胞来维持产生激素的细胞。特别是，表达 S100 β 蛋白（一种钙结合蛋白）的细胞被假设为垂体细胞资源。积累的数据表明，S100 β 阳性细胞包含异质群体，其中一些在体内确实表现出干细胞/祖细胞特征。因此，我们通过对在S100β启动子控制下表达增强型绿色荧光蛋白（EGFP）的转基因大鼠进行体内和体外实验，检测S100β阳性细胞是否具有分化为内分泌细胞的能力。垂体免疫组织化学证实，一些 S100 β 阳性细胞表达 SOX2（SRY [性别决定区 Y]-box 2）并具有增殖活性。通过延时显微镜观察分散的前叶细胞，然后进行激素和垂体转录因子 1 (PIT1) 的免疫染色。首先，用抗 SOX2 抗体对分散的前叶细胞进行免疫染色。 S100 β 蛋白与 SOX2 共定位（约 89%）。尽管追踪到的 134 个 S100 β 阳性细胞中有 44 个处于增殖状态，但对任何激素呈阴性，但有 14 个细胞对一种垂体激素和/或 PIT1 呈阳性，这证实了所有类型的激素产生细胞的存在。值得注意的是，培养过程中两个激素阳性细胞中出现了 GFP 荧光。另一方面，我们观察到，尽管最初呈阳性，但在延时研究结束时，产生激素的细胞对 S100 β 不呈阳性。这些发现表明，从前叶培养的 S100 β 阳性细胞能够发育成产生激素的细胞，尽管这种情况发生相对较少。
Some non-endocrine cells in the pituitary anterior lobe are responsible for providing stem/progenitor cells to maintain hormone-producing cells. In particular, cells expressing S100 beta protein, a calcium-binding protein, have been hypothesized to be a pituitary cell resource. Accumulating data have revealed that S100 beta-positive cells comprise heterogeneous populations and some of them certainly show stem/progenitor characteristics in vivo. Hence, we examine whether S100 beta-positive cells have the capacity to differentiate into endocrine cells, by means of in vivo and in vitro experiments on transgenic rats expressing enhanced green fluorescent protein (EGFP) under the control of the S100 beta promoter. Immunohistochemistry of the pituitary confirmed that some S100 beta-positive cells expressed SOX2 (SRY [sex-determining region Y]-box 2) and had proliferative activity. Dispersed anterior lobe cells were observed by time-lapse microscopy, followed by immunostaining for hormone and pituitary-transcription-factor1 (PIT1). First, the dispersed anterior lobe cells were immunostained by an antibody against SOX2. S100 beta-protein co-localizes with SOX2 (about 89 %). Although 44 of 134 S100 beta-positive cells traced were proliferative but negative to any hormones, 14 cells were positive for one of the pituitary hormones and/or PIT1, confirming the presence of all types of hormone-producing cells. Notably, GFP-fluorescence appeared in two hormone-positive cells during culture. On the other hand, we observed hormone-producing cells that were not positive for S100 beta at the end of the time-lapse study, despite being initially positive. These findings suggest that S100 beta-positive cells cultured from the anterior lobe are capable of developing into hormone-producing cells, although this happens relatively infrequently.