Elucidation and Application for Mechanism of Differentiation in Germ Cells from Conditionally Immortalized Leydig and Sertoli Cell Lines from Transgenic Mice

建立了支持细胞系TTE3和间质细胞系TTE1。他们的性格已经被报道出来了。在TTE1和TTE3细胞中，细胞生长在非允许温度下受到抑制。这表明细胞没有转化。我们对它们进行了细胞-细胞粘附的形态学检查。在TTE3细胞中，它们在非允许温度下形成更紧密的连接。在TTE1细胞中，它们不形成紧密连接。在TTE3中，细胞表达编码钢铁因子、抑制素-α、转铁蛋白、促卵泡激素受体和硫酸糖蛋白-2的mrna。在细胞质中观察到波形蛋白，在细胞边界上观察到闭塞带-1。有趣的是，在非允许温度下，转铁蛋白和封闭带-1（紧密连接成分）的表达水平显著升高。d.逆转录聚合酶链式反应（RT-PCR）分析表明，TTE1细胞表达17β-HSD型和3型以及抑制素-α的mrna。未来，我们希望建立一个模型来解决关于间质细胞、支持细胞和生殖细胞相互作用性质的重要问题。为了鉴定支持细胞和间质细胞分化过程中的差异表达基因，我们对TTE3细胞和TTE1细胞在允许和非允许温度下培养进行了微阵列分析。在TTE3中，分析的1080个基因（IntelliGene microarray）中，29个基因的表达水平发生了变化，其中20个基因表达水平升高，其余9个基因表达水平降低。在TTE1中，1081个基因（Atlass微阵列）中，31个基因的表达水平发生了变化，其中24个基因表达水平升高，其余7个基因表达水平降低。分化条件下，Tie2表达量上调13.5倍，是变化最大的转录本。下一步，我们想要研究这些与细胞-细胞粘附（或紧密连接）相关的基因，它们的表达是否会改变。少

The Sertoli cell line, TTE3,and Leydig cell line, TTE1, were established. Their characters are already reported. In TTE1 and TTE3 cells, cell growth was suppressive at nonpermissive temperature. This indicated that the cells were not transformed.We investigated their morphological examination for cell-cell adhesion. In TTE3 cells, they formed more tight-junction in nonpermissive temperature. In TTE1 cells they did not form tight-junction.In TTE3, the cells expressed mRNAs encoding steel factor, inhibin-α, transferrin, follicle stimulating hormone receptor and sulfated glycoprotein-2. Moreover, vimentin and zonula occludens-1 were observed in the cytoplasm and on the boundaries of the cells, respectively. Interestingly expression levels of transferrin and zonula occludens-1(tight junction component) were significantly elevated nonpermissive temperature.In TTE1, alkaline phosphatase and 3β-hydroxysteroid dehydrogenase(HSD) activities or expression of cytokeratin and vimentin were observe … More d. Reverse transcription-polymerase chain reaction(RT-PCR) analysis indicated that TTE1 cells expressed mRNAs encoding 17β-HSD typesl and 3, and inhibin-α.In future, we want to make a model for addressing important questions regarding the nature of Leydig cells, Sertoli cells, and germ cell interactions.To identify differentially expressed genes in the process of Sertoli cell and Leydig cell differentiation, we carried out microarray analysis of TTE3 cells and TTE1 cells cultured at permissive and nonpermissive temperatures. In TTE3, of the 1080 genes(IntelliGene microarray) analyzed, the level of 29 genes were changed, with 20 genes showing increased levels of expression and the remaining 9 genes showing decreased levels. In TTE1, of the 1081 genes(Atlass microarray), the levels of 31 genes were changed, with 24 genes showing increased levels of expression and the remaining 7 genes showing decreased levels. Tie2 was the most changed transcript, with a 13.5-fold upregulation under the differentiated condition.In the next step, we want to study these genes related to cell-cell adhesion(or tight junction) whether their expression will change or not. Less