Abundance of Early Embryonic Primordial Germ Cells Promotes Zebrafish Female Differentiation as Revealedby Lifetime Labeling of Germline
Abundance of Early Embryonic Primordial Germ Cells Promotes Zebrafish Female Differentiation as Revealedby Lifetime Labeling of Germline
复制标题
种系终生标记揭示早期胚胎原始生殖细胞的丰富性促进斑马鱼雌性分化
DOI：
10.1007/s10126-019-09874-1
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发表时间：
2019
影响因子：
3
通讯作者：
Sun Yonghua
中科院分区：
文献类型：
--
作者：
Ye Ding;Zhu Lin;Zhang Qifeng;Xiong Feng;Wang Houpeng;Wang Xiaosi;He Mudan;Zhu Zuoyan;Sun Yonghua
硬骨鱼的性别分化很大程度上取决于未分化生殖细胞的数量。在这里，我们描述了一种新的转基因斑马鱼品系Tg(piwil1:egfp-UTRnanos3)ihb327Tg的产生和特性，该品系可以特异性标记生殖细胞的整个生命周期，即从处于屏障期的原始生殖细胞（PGCs）到卵巢中的卵原细胞和早期的卵母细胞，再到睾丸中的精原细胞、精母细胞和精细胞的早期阶段。利用该转基因系，我们仔细观察了从胚胎早期到幼年期PGCs的数量以及卵巢和睾丸的分化过程。早在受精后1天（dpf）， PGCs的数量就开始变化。有趣的是，PGCs含量高的胚胎主要发育为雌性，而PGCs含量低的胚胎主要发育为雄性。通过瞬时过表达和转基因诱导pgc特异性巴基球（buc），我们进一步证明了在胚胎期诱导丰富的PGCs促进后期卵巢分化和雌性发育。由此，我们获得了一个理想的转基因品系（piwil1:egfp-UTRnanos3） ihb327tg1，该品系可以对斑马鱼生殖系进行终身可视化，并利用该品系对硬骨鱼生殖细胞发育和性腺分化进行了研究，证明胚胎期PGC数量的增加促进了雌性分化。
Teleost sex differentiation largely depends on the number of undifferentiated germ cells. Here, we describe the generation and characterization of a novel transgenic zebrafish line,Tg(piwil1:egfp-UTRnanos3)ihb327Tg, which specifically labels the whole lifetime of germ cells, i.e., from primordial germ cells (PGCs) at shield stage to the oogonia and early stage of oocytes in the ovary and to the early stage of spermatogonia, spermatocyte, and spermatid in the testis. By using this transgenic line, we carefully observed the numbers of PGCs from early embryonic stage to juvenile stage and the differentiation process of ovary and testis. The numbers of PGCs became variable at as early as 1 day post-fertilization (dpf). Interestingly, the embryos with a high amount of PGCs mainly developed into females and the ones with a low amount of PGCs mainly developed into males. By using transient overexpression and transgenic induction of PGC-specificbucky ball(buc), we further proved that induction of abundant PGCs at embryonic stage promoted later ovary differentiation and female development. Taken together, we generate an ideal transgenic lineTg(piwil1:egfp-UTRnanos3)ihb327Tgwhich can visualize zebrafish germline for a lifetime, and we have utilized this line to study germ cell development and gonad differentiation of teleost and to demonstrate that the increase of PGC number at embryonic stage promotes female differentiation.