Laboratory of Veterinary Physiology, School of Veterinary Medicine, Faculty of Veterinary Science, Nippon Veterinary and Life Science University, Tokyo, Japan.
Department of Anatomy & Histology, Faculty of Veterinary Medicine and Animal Science, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh.
J Histochem Cytochem. 2021 Apr;69(4):257-270. doi: 10.1369/0022155421991629. Epub 2021 Feb 10.
A well-known putative tumor suppressor WW domain-containing oxidoreductase (Wwox) is highly expressed in hormonally regulated tissues and is considered important for the normal development and function of reproductive organs. In this study, we investigated the cellular and subcellular localization of Wwox in normal testes during postnatal days 0-70 using Western blotting and immunohistochemistry. Wwox is expressed in testes at all ages. Immunohistochemistry showed that fetal-type and adult-type Leydig cells, immature and mature Sertoli cells, and germ cells (from gonocytes to step 17 spermatids) expressed Wwox except peritubular myoid cells, step 18-19 spermatids, and mature sperm. Wwox localized diffusely in the cytoplasm with focal intense signals in all testicular cells. These signals gradually condensed in germ cells with their differentiation and colocalized with giantin for cis-Golgi marker and partially with golgin-97 for trans-Golgi marker. Biochemically, Wwox was detected in isolated Golgi-enriched fractions. But Wwox was undetectable in the nucleus. This subcellular localization pattern of Wwox was also confirmed in single-cell suspension. These findings indicate that Wwox is functional in most cell types of testis and might locate into Golgi apparatus via interaction with Golgi proteins. These unique localizations might be related to the function of Wwox in testicular development and spermatogenesis.
一个众所周知的假定肿瘤抑制因子 WW 结构域氧化还原酶（Wwox）在激素调节组织中高度表达，被认为对生殖器官的正常发育和功能很重要。在这项研究中，我们使用 Western blot 和免疫组织化学方法研究了 Wwox 在出生后 0-70 天的正常睾丸中的细胞和亚细胞定位。Wwox 在所有年龄段的睾丸中都有表达。免疫组织化学显示，胎儿型和成年型 Leydig 细胞、未成熟和成熟的 Sertoli 细胞以及生殖细胞（从精原细胞到第 17 步精子）都表达了 Wwox，除了小管周肌样细胞、第 18-19 步精子和成熟精子。Wwox 在所有睾丸细胞的细胞质中弥散表达，具有局灶性强信号。这些信号随着生殖细胞的分化而逐渐浓缩，并与高尔基标志物 giantin 共定位，部分与高尔基标志物 golgin-97 共定位。生化分析表明，Wwox 存在于分离的富含高尔基的级分中。但是，Wwox 在核内不可检测。这种 Wwox 的亚细胞定位模式也在单细胞悬液中得到了证实。这些发现表明，Wwox 在睾丸的大多数细胞类型中具有功能，并且可能通过与高尔基蛋白的相互作用定位到高尔基器中。这些独特的定位可能与 Wwox 在睾丸发育和精子发生中的功能有关。
