Kraszucka K, Burfeind P, Nayernia K, Köhler M, Schmid M, Yaylaoglu M, Engel W
Institute of Human Genetics, University of Göttingen, Göttingen, Germany.
Mol Reprod Dev. 1999 Nov;54(3):232-43. doi: 10.1002/(SICI)1098-2795(199911)54:3<232::AID-MRD4>3.0.CO;2-F.
There is considerable evidence that germ cells, mainly spermatocytes and spermatids, contribute to the regulation of Sertoli cell activity. We developed an in vitro system to investigate the genes involved in Sertoli cell-germ cell interaction in the mouse by using the differential mRNA display technique. One of the isolated differentially expressed genes, named calgizzarin, belongs to the family of S100 calcium-binding proteins and shows a decreased expression in Sertoli cell-germ cell cocultures compared to cultured Sertoli cells alone. Calgizzarin is expressed in all adult tissues examined, including testis and ovary; however, a high mRNA level for calgizzarin in mouse testis is maintained until day 15 of postnatal development and then declines dramatically, whereas the expression pattern in the ovary remains constantly high. Furthermore, Northern blot studies on testicular RNA from different mouse mutants with defects in spermatogenesis revealed that high levels of calgizzarin transcripts can only be detected in testes of mouse mutants with either no germ cells or primary spermatocytes, but only weak signals for calgizzarin are observed in testes of mutants containing spermatids. In addition, using both RT-PCR analysis and whole-mount in situ hybridization on dissected gonads it was demonstrated that mouse calgizzarin expression starts at 13.5 dpc in the prenatal male gonad and at 16.5 dpc in the embryonic ovary, respectively. The mouse calgizzarin gene was localized on mouse chromosome 5, region E-F. Taken together, our results indicate that calgizzarin expression could be repressed by factors originated from pachytene spermatocytes and/or spermatids.
有大量证据表明，生殖细胞，主要是精母细胞和精子细胞，参与支持细胞活性的调节。我们开发了一种体外系统，通过使用差异mRNA显示技术来研究小鼠中参与支持细胞 - 生殖细胞相互作用的基因。其中一个分离出的差异表达基因，名为钙结合蛋白，属于S100钙结合蛋白家族，与单独培养的支持细胞相比，在支持细胞 - 生殖细胞共培养中表达降低。钙结合蛋白在所有检测的成年组织中均有表达，包括睾丸和卵巢；然而，小鼠睾丸中钙结合蛋白的高mRNA水平一直维持到出生后第15天，然后急剧下降，而卵巢中的表达模式则一直保持高水平。此外，对不同精子发生缺陷的小鼠突变体睾丸RNA进行的Northern印迹研究表明，只有在没有生殖细胞或初级精母细胞的小鼠突变体睾丸中才能检测到高水平的钙结合蛋白转录本，而在含有精子细胞的突变体睾丸中仅观察到微弱的钙结合蛋白信号。此外，通过对解剖性腺进行RT-PCR分析和全胚胎原位杂交表明，小鼠钙结合蛋白的表达分别在产前雄性性腺的13.5 dpc和胚胎卵巢的16.5 dpc开始。小鼠钙结合蛋白基因定位于小鼠第5号染色体的E - F区域。综上所述，我们的结果表明，钙结合蛋白的表达可能受到来自粗线期精母细胞和/或精子细胞的因子的抑制。
