Department of Marine Biosciences, Tokyo University of Marine Science and Technology, Tokyo, Japan.
Biol Reprod. 2010 Oct;83(4):597-606. doi: 10.1095/biolreprod.109.082081. Epub 2010 Jun 10.
In mammals, several cell surface molecular markers have been characterized in order to identify the mitotic germ cells. However, little is known in fish about their cell surface antigen. In this study, we identified lymphocyte antigen 75 (Ly75/CD205) as a germ cell-specific cell surface marker by combination expressed sequence tag analysis of purified type A spermatogonia (A-SG) from immature testis, in silico prediction of membrane proteins, and expression studies. The ly75 transcripts were abundant in the testis and gills, and weak signals were detected in the head kidney and brain. In addition, ly75 mRNA was predominantly localized in the primordial germ cells of newly hatched embryos, A-SG in testis, oogonia, and chromatin nucleolus-stage oocytes in the ovary. In contrast, ly75 mRNA was not detected in spermatocytes, spermatids, spermatozoa, vitellogenic oocytes, or gonadal somatic cells from either males or females. The expression profile of Ly75 protein was similar to that of the mRNA. Furthermore, identification of various fish homologs of ly75 confirmed that their amino acid sequences are well conserved. Therefore, Ly75 may be appropriate for use as a versatile cell surface marker for mitotic germ cells in fish.
在哺乳动物中,已经鉴定了几种细胞表面分子标记物,以鉴定有丝分裂生殖细胞。然而,鱼类中关于它们的细胞表面抗原知之甚少。在这项研究中,我们通过对未成熟睾丸中纯化的 A 型精原细胞(A-SG)进行组合表达序列标签分析、膜蛋白的计算机预测和表达研究,鉴定了淋巴细胞抗原 75(Ly75/CD205)作为一种生殖细胞特异性细胞表面标记物。ly75 转录本在睾丸和鳃中丰富,在头肾和脑中检测到较弱的信号。此外,ly75 mRNA 主要定位于刚孵化的胚胎中的原始生殖细胞、睾丸中的 A-SG、卵母细胞中的卵原细胞以及卵巢中的染色质核仁期卵母细胞。相反,ly75 mRNA 未在精母细胞、精细胞、精子、卵黄发生卵母细胞或雄性或雌性的性腺体细胞中检测到。Ly75 蛋白的表达谱与 mRNA 相似。此外,鉴定各种鱼类 ly75 的同源物证实它们的氨基酸序列高度保守。因此,Ly75 可能适合用作鱼类有丝分裂生殖细胞的通用细胞表面标记物。