Dipartimento di Medicina Sperimentale, Sez. Fisiologia Umana e Funzioni Biologiche Integrate 'F. Bottazzi', Università degli Studi della Campania 'Luigi Vanvitelli' via Costantinopoli, 16-80138 - Napoli, Italy.
Dipartimento di Biologia, Università di Napoli 'Federico II, via Cinthia', 21-80126 - Napoli, Italy.
Zygote. 2020 Dec;28(6):441-446. doi: 10.1017/S0967199420000301. Epub 2020 Jul 29.
In 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.
在本研究中，我们旨在寻找哺乳动物精子发生过程中涉及的新基因，首次从大鼠睾丸中分离出一个部分 cDNA 克隆，该克隆编码 EH 结构域结合蛋白 1 样 1（Ehbp1l1），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 端 C2 样结构域、CH 结构域和 C 端二价 Mical/EHBP Rab 结合（bMERB）结构域，这些结构域在脊椎动物中是进化保守的。我们发现 Ehbp1l1 mRNA 在几种大鼠组织中表达，包括肝脏、肠道、肾脏，在睾丸发育过程中也有表达，在 12 月龄动物的睾丸中表达水平较高。有趣的是，原位杂交实验显示 Ehbp1l1 特异性表达于 I 型和 II 型精母细胞，这一结果通过对从不同睾丸细胞类型富集的总 RNA 进行 RT-PCR 得到了验证。由于 EHBP1l1 已被描述为与囊泡运输到肌动蛋白细胞骨架有关，并且是小 GTPase Rab8 的效应物，我们假设它可能参与细胞骨架重塑和囊泡从高尔基网络到顶质膜的分拣调节。我们的研究结果提供了对精子发生分化过程分子机制的更好理解；Ehbp1l1 也可用作睾丸活性的新标志物。
