Syed V, Hecht N B
Department of Biology, Tufts University, Medford, Massachusetts 02155, USA.
Mol Reprod Dev. 1997 Aug;47(4):380-9. doi: 10.1002/(SICI)1098-2795(199708)47:4<380::AID-MRD4>3.0.CO;2-G.
To better understand the molecular interactions between somatic and germ cells in the mammalian testis, we have begun to analyze with mRNA differential display changes in gene expression induced by coculturing rat Sertoli cells and germ cells. We have identified 10 cDNAs that are either down-regulated or up-regulated in cocultures of germ cells and Sertoli cells. Three genes expressed in Sertoli cells and three genes expressed in germ cells were down-regulated in Sertoli cell-germ cell cocultures, whereas four genes were up-regulated in the cocultures. Northern blot analysis was used to establish the expression pattern of the mRNAs encoded by the cDNAs and to define the sizes of the differentially expressed mRNAs. Sequence analysis of the cDNAs and computer searches against the GenBank and EMBL DNA databases were used to relate the ten cDNAs to known genes. Of the three Sertoli cell cDNAs, one appeared identical to transferin, while the other two shared regions of similarity to an endoplasmic reticulum stress protein and to a pro-alpha 2 XI collagen, respectively. The three germ cell cDNAs shared sequences with fibronectin, with a basic fibroblast growth factor receptor and with an IgG gamma 2b, respectively. The four cDNAs that were up-regulated in the Sertoli-germ cell cocultures showed similarity to an isoform of casein kinase 1 delta, to an epidermal growth factor, to a statin-related protein, and to an integral membrane glycoprotein. These data demonstrate that a number of specific genes are up- and down-regulated when germ cells and Sertoli cells are cocultured, and suggest these genes are important in cell to cell communication during spermatogenesis.
为了更好地理解哺乳动物睾丸中体细胞与生殖细胞之间的分子相互作用,我们已开始运用mRNA差异显示技术,分析大鼠支持细胞与生殖细胞共培养所诱导的基因表达变化。我们鉴定出了10个在生殖细胞与支持细胞共培养中表达下调或上调的cDNA。在支持细胞-生殖细胞共培养中,支持细胞中表达的3个基因和生殖细胞中表达的3个基因表达下调,而共培养中有4个基因表达上调。采用Northern印迹分析来确定由这些cDNA编码的mRNA的表达模式,并确定差异表达mRNA的大小。对这些cDNA进行序列分析,并针对GenBank和EMBL DNA数据库进行计算机检索,以将这10个cDNA与已知基因关联起来。在支持细胞的3个cDNA中,一个似乎与转铁蛋白相同,而另外两个分别与一种内质网应激蛋白和一种前α2 XI型胶原蛋白有相似区域。这3个生殖细胞cDNA分别与纤连蛋白、碱性成纤维细胞生长因子受体和IgGγ2b有共同序列。在支持细胞-生殖细胞共培养中上调的4个cDNA与酪蛋白激酶1δ的一种同工型、一种表皮生长因子、一种他汀类相关蛋白和一种整合膜糖蛋白有相似性。这些数据表明,当生殖细胞与支持细胞共培养时,许多特定基因的表达会发生上调和下调,这表明这些基因在精子发生过程中的细胞间通讯中很重要。