Abstract：
Spermatogenesis is a complicated process that includes spermatogonia differentiation, spermatocytes meiosis, spermatids spermiogenesis and final release of spermatozoa. Actin-related protein 3 (Arp3) and epidermal growth factor receptor pathway substrate 8 (Eps8) are two actin binding proteins that regulate cell adhesion in seminiferous tubules during mammalian spermatogenesis. However, the functions of these two proteins during spermatogenesis in nonmammalian species, especially Crustacea, are still unknown. Here, we cloned es-Arp3 and es-Eps8 from the testis of Chinese mitten crab Eriocheir sinensis. es-Arp3 and es-Eps8 were located in spermatocytes, spermatids and spermatozoa. Knockdown of es-Arp3 and es-Eps8 in vivo caused morphological changes to seminiferous tubules including delayed spermatozoa release, shedding of germ cells and vacuoles. Filamentous-actin (F-actin) filaments network was disorganized due to deficiency of es-Arp3 and es-Eps8. Accompanying this, four junctional proteins (α-catenin, β-catenin, pinin and ZO1) displayed abnormal expression levels as well as penetrating biotin signals in seminiferous tubules. We also used the Arp2/3 complex inhibitor CK666 to block es-Arp3 activity and supported es-Arp3 knockdown results. In summary, our study demonstrated for the first time that es-Arp3 and es-Eps8 are important for spermatogenesis via regulating microfilament-mediated cell adhesion in Eriocheir sinensis.
摘要：
精子发生是一个复杂的过程，包括精原细胞分化，精母细胞减数分裂,精子细胞精子发生和精子的最终释放。肌动蛋白相关蛋白3（Arp3）和表皮生长因子受体途径底物8（Eps8）是两种肌动蛋白结合蛋白，可在哺乳动物精子发生过程中调节生精小管中的细胞粘附。然而,这两种蛋白质在非哺乳动物物种精子发生过程中的功能，尤其是甲壳动物,仍然未知。这里,我们从中华绒螯蟹的睾丸中克隆了es-Arp3和es-Eps8。es-Arp3和es-Eps8位于精母细胞中，精子和精子。在体内敲除es-Arp3和es-Eps8会引起生精小管的形态变化，包括精子释放延迟，生殖细胞和空泡脱落。由于es-Arp3和es-Eps8的缺乏，丝状肌动蛋白（F-肌动蛋白）细丝网络混乱。伴随着这个,四种连接蛋白(α-连环蛋白，β-连环蛋白，pinin和ZO1）在生精小管中显示异常表达水平以及穿透生物素信号。我们还使用Arp2/3复合物抑制剂CK666来阻断es-Arp3活性并支持es-Arp3敲低结果。总之,我们的研究首次证明，es-Arp3和es-Eps8通过调节微丝介导的细胞粘附对中华绒螯蟹的精子发生很重要。
