Evidence for cross-talk between Sertoli and germ cells using selected cathepsins as markers.
Evidence for cross-talk between Sertoli and germ cells using selected cathepsins as markers.
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使用选定的组织蛋白酶作为标记物支持支持细胞和生殖细胞之间串扰的证据。
作者：
Sanny S. W. Chung;L. Zhu;Monteros;B. Silvestrini;Will M. Lee;Cheng Cy
为了研究蛋白酶是否可能参与细胞迁移和/或精子形成时，发展中的生殖细胞易位在生精上皮在精子发生过程中，原位杂交被用来本地化的信使RNA（mRNA）转录的组织蛋白酶L，D，和S在上皮细胞在不同阶段的生精周期在大鼠。组织蛋白酶L mRNA被发现几乎只定位在上皮的基底层附近。在精子形成前的周期的第VI和第VII阶段，组织蛋白酶L mRNA的信号是如此强烈，以至于它形成了一个完整的黑暗沉淀物附近的基膜包围整个小管。在第八阶段，组织蛋白酶L的表达被完全废除，并没有染色的组织蛋白酶L的mRNA被视为在上皮细胞。组织蛋白酶D和S的mRNA被发现附近的基底层，发现与他们的本地化支持细胞和初级精母细胞在几乎所有阶段一致，但在第七-IX和第七-VIII阶段的周期，分别达到峰值，在精子形成之前和期间的时间。这些结果说明这些蛋白酶可能参与促进生殖细胞运动和精子形成。为了检查生殖细胞是否表达这些组织蛋白酶基因中的任何一种，通过Percoll梯度离心法从15日龄大鼠睾丸中分离纯度大于95%的精母细胞进行逆转录-聚合酶链反应。结果发现，初级精母细胞表达多种组织蛋白酶基因，包括组织蛋白酶B、C、D、H、L和S。此外，组织蛋白酶L的表达，从15日龄大鼠（主要是精母细胞和精原细胞）分离的生殖细胞可以刺激支持细胞富集培养基中的剂量依赖性的方式，但不是生殖细胞条件培养基。这些结果表明，支持细胞可以调节生殖细胞的功能。此外，这些结果表明，生殖细胞可能在整体睾丸蛋白酶表达中发挥积极作用。此外，我们提出的证据表明，支持细胞和生殖细胞之间存在串扰，因为组织蛋白酶L在每种细胞类型中的表达通过可溶性因子或细胞-细胞接触相互调节。
To examine whether proteases are possibly involved in cellular migration and/or spermiation when developing germ cells translocate across the seminiferous epithelium during spermatogenesis, in situ hybridization was used to localize messenger RNA (mRNA) transcripts of cathepsin L, D, and S in the epithelium at different stages of the spermatogenic cycle in the rat. Cathepsin L mRNA was found to localize almost exclusively near the basal lamina of the epithelium. At stages VI and VII of the cycle before spermiation, the signal of cathepsin L mRNA was so intense that it formed a complete dark precipitate near the basal lamina encircling the entire tubule. At stage VIII, the expression of cathepsin L was completely abolished, and no staining of cathepsin L mRNA was seen in the epithelium. The mRNA of cathepsin D and S was found near the basal lamina, a finding consistent with their localization in Sertoli cells and possibly primary spermatocytes in almost all stages, but peaked at stages VII-IX and VII-VIII of the cycle, respectively, at the time before and during spermiation. These results illustrate the possible involvement of these proteases in facilitating germ cell movement and spermiation. To examine whether germ cells express any of these cathepsin genes, spermatocytes with a purity of greater than 95% were isolated from 15-day-old rat testes by Percoll gradient centrifugation for reverse transcriptase-polymerase chain reaction. It was found that primary spermatocytes expressed multiple cathepsin genes, including cathepsin B, C, D, H, L, and S. Furthermore, the expression of cathepsin L by germ cells isolated from 15-day-old rats (largely spermatocytes and spermatogonia) can be stimulated by Sertoli cell-enriched culture medium in a dose-dependent manner, but not by germ cell-conditioned medium. These results reveal that germ cell function can be regulated by Sertoli cells. Moreover, these results suggest that germ cells may play an active role in the overall testicular protease expression. Also, we present evidence suggesting there is cross-talk between Sertoli and germ cells, since the expression of cathepsin L in each cell type is regulated by one another via either soluble factors or cell-cell contact.
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