Ozaki Yuichi, Saito Kenji, Shinya Minori, Kawasaki Toshihiro, Sakai Noriyoshi
Genetic Strains Research Center, National Institute of Genetics, Mishima 411-8540, Japan.
Gene Expr Patterns. 2011 Jun-Jul;11(5-6):309-15. doi: 10.1016/j.gep.2011.03.002. Epub 2011 Mar 21.
Recent studies in mammals have revealed the heterogeneity of spermatogonial populations which contain differentiated and undifferentiated cells that further divide into actual stem cells and potential stem cells. In fish however, there are no functional definitions, and very few molecular markers, for germ cells. In our present study, specific antibodies were raised against Sycp3, Plzf and Cyclin B3 in zebrafish and then used to determine the localization of these proteins in the testis. We wished to confirm whether these molecules were potential markers for spermatocytes and spermatogonia. Immunohistochemical observations revealed that Sycp3 is specifically localized in spermatocytes in typical nuclear patterns at each meiotic stage. Plzf was found to be localized in the nucleus of both type A and type B spermatogonia until the 8-cell clone, similar to the pattern in Plzf-positive A(single)-A(aligned) undifferentiated spermatogonia in rodents. In addition to Plzf, the localization of Cyclin B3 was predominantly detected in the nuclei of type A and early type B spermatogonia until the 16-cell clone. Additionally, Cyclin B3 protein signals were detected in germ cells in large cysts, possibly corresponding to spermatocytes at the preleptotene stage. Our present data thus show that these molecules have properties that will enable their use as markers of spermatocytes and early spermatogonia in zebrafish.
最近在哺乳动物中的研究揭示了精原细胞群体的异质性，其中包含已分化和未分化的细胞，这些细胞进一步分为实际干细胞和潜在干细胞。然而，在鱼类中，生殖细胞没有功能定义，分子标记也很少。在我们目前的研究中，针对斑马鱼中的Sycp3、Plzf和细胞周期蛋白B3制备了特异性抗体，然后用于确定这些蛋白质在睾丸中的定位。我们希望确认这些分子是否是精母细胞和精原细胞的潜在标记物。免疫组织化学观察表明，Sycp3在每个减数分裂阶段以典型的核模式特异性定位于精母细胞中。发现Plzf在8细胞克隆之前定位于A型和B型精原细胞的细胞核中，类似于啮齿动物中Plzf阳性的A（单）-A（排列）未分化精原细胞的模式。除了Plzf，细胞周期蛋白B3的定位在16细胞克隆之前主要在A型和早期B型精原细胞的细胞核中检测到。此外，在大囊肿中的生殖细胞中检测到细胞周期蛋白B3蛋白信号，可能对应于细线前期的精母细胞。因此，我们目前的数据表明，这些分子具有可作为斑马鱼精母细胞和早期精原细胞标记物的特性。
