Characterization of lymphocyte antigen 75 (Ly75/CD205) as a potential cell-surface marker on spermatogonia in Pacific bluefin tuna Thunnus orientalis
Characterization of lymphocyte antigen 75 (Ly75/CD205) as a potential cell-surface marker on spermatogonia in Pacific bluefin tuna Thunnus orientalis
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DOI：
10.1007/s12562-012-0501-9
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发表时间：
2012-07-01
影响因子：
1.9
通讯作者：
Yoshizaki, Goro
中科院分区：
文献类型：
--
作者：
Nagasawa, Kazue;Miwa, Misako;Yoshizaki, Goro
在以东方金枪鱼（Thunnus orientalis）为供体的精原细胞移植中，精原细胞的富集有望提高移植效率。虽然它是可取的，以建立一个蓝鳍金枪鱼SG富集程序使用细胞表面标记，生殖细胞特异性细胞表面标记尚未确定日期。我们以前发现Ly 75是虹鳟鱼有丝分裂生殖细胞特异性的细胞表面标记，并且其氨基酸序列在各种硬骨鱼中高度保守。因此，ly 75基因是蓝鳍金枪鱼SG的一个很好的候选细胞表面标记。在这项研究中，蓝鳍金枪鱼ly 75同源物的克隆和进一步使用作为生殖细胞特异性细胞表面标记的特点。在成年组织中，在肝脏、幽门盲囊和睾丸中检测到高水平的ly 75转录本。原位杂交分析表明，ly 75 mRNA主要定位于A型精原细胞（A-SG），包括单个A-SG含有可移植的生殖细胞。相反，在睾丸的精母细胞、精子细胞或性腺体细胞中未检测到ly 75 mRNA。Ly 75蛋白表达谱与mRNA表达谱相似。因此，Ly 75适合作为蓝鳍金枪鱼SG的细胞表面标记。
In spermatogonial transplantation using Pacific bluefin tuna Thunnus orientalis as a donor, enrichment of spermatogonia (SG) is expected to facilitate high colonization efficiency. Although it is desirable to establish a bluefin tuna SG enrichment procedure using cell-surface markers, a germ cell-specific cell-surface marker has not been identified to date. We previously found that Ly75 is a mitotic germ cell-specific cell-surface marker in rainbow trout, and that its amino-acid sequences are highly conserved in various teleosts. Thus, the ly75 gene is an excellent candidate cell-surface marker of SG in bluefin tuna. In this study, the bluefin tuna ly75 homolog was cloned and characterized for further use as a germ cell-specific cell-surface marker. In adult tissues, high levels of ly75 transcripts were detected in the liver, pyloric caeca, and testis. In situ hybridization analyses showed that ly75 mRNA was predominantly localized in type-A spermatogonia (A-SG), including single A-SG that contain transplantable germ cells. In contrast, ly75 mRNA was not detected in spermatocytes, spermatids, or gonadal somatic cells in testis. The expression profiles of Ly75 protein were similar to those of the mRNA. Therefore, Ly75 is appropriate for use as a cell-surface marker of SG in bluefin tuna.