SummaryIn this paper, with the aim to find new genes involved in mammalian spermatogenesis, we isolated, for the first time in the rat testis, a partial cDNA clone that encoded EH domain binding protein 1-like 1 (Ehbp1l1), a protein that has a single calponin homology domain (CH). Bioinformatic analysis showed that EHBP1l1 contains three domains: the N-terminal C2-like, the CH and the C-terminal bivalent Mical/EHBP Rab binding (bMERB) domains, which are evolutionarily conserved in vertebrates. We found that Ehbp1l1 mRNA was expressed in several rat tissues, including the liver, intestine, kidney and also in the testis during its development, with a higher level in testis from 12-month-old animals. Interestingly, in situ hybridization experiments revealed that Ehbp1l1 is specifically expressed by types I and II spermatocytes, this result was validated by RT-PCR performed on total RNA obtained from enriched fractions of different testicular cell types. As EHBP1l1 has been described as linked to vesicular transport to the actin cytoskeleton and as an effector of the small GTPase Rab8, we hypothesized that it could participate both in cytoskeletal remodelling and in the regulation of vesicle sorting from the trans-Golgi network to the apical plasma membrane. Our findings provide a better understand of the molecular mechanisms of the differentiation process of spermatogenesis; Ehbp1l1 may also be used as a new marker of testicular activity.

中文翻译：

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EH 结构域结合蛋白 1 样 1 (EHBP1L1) 是一种具有 calponin 同源结构域的蛋白质，在大鼠睾丸中表达

摘要在本文中，为了寻找与哺乳动物精子发生有关的新基因，我们首次在大鼠睾丸中分离出编码 EH 结构域结合蛋白 1-like 1 (Ehbp1l1) 的部分 cDNA 克隆，该蛋白具有单个钙调蛋白同源结构域 (CH)。生物信息学分析表明，EHBP1l1 包含三个结构域：N 末端 C2 样、CH 和 C 末端二价 Mical/EHBP Rab 结合 (bMERB) 结构域，它们在脊椎动物中是进化上保守的。我们发现 Ehbp1l1 mRNA 在其发育过程中在几种大鼠组织中表达，包括肝、肠、肾和睾丸，在 12 个月大的动物的睾丸中表达水平更高。有趣的是，原位杂交实验表明，Ehbp1l1 由 I 型和 II 型精母细胞特异性表达，这一结果通过对从不同睾丸细胞类型的富集部分获得的总 RNA 进行的 RT-PCR 验证。由于 EHBP1l1 已被描述为与向肌动蛋白细胞骨架的囊泡转运有关，并且作为小 GTPase Rab8 的效应器，我们假设它可以参与细胞骨架重塑和囊泡分选的调节。反式- 顶端质膜的高尔基网络。我们的研究结果有助于更好地理解精子发生分化过程的分子机制；Ehbp1l1 也可用作睾丸活动的新标志物。