CRISPR/Cas9-induced disruption of wt1a and wt1b reveals their different roles in kidney and gonad development in Nile tilapia
CRISPR/Cas9-induced disruption of wt1a and wt1b reveals their different roles in kidney and gonad development in Nile tilapia
复制标题
CRISPR/Cas9 诱导的 wt1a 和 wt1b 破坏揭示了它们在尼罗罗非鱼肾脏和性腺发育中的不同作用
DOI：
10.1016/j.ydbio.2017.05.017
复制
发表时间：
2017-08-01
影响因子：
2.7
通讯作者：
Wang, Deshou
中科院分区：
文献类型：
--
作者：
Jiang, Dongneng;Chen, Jinlin;Wang, Deshou
Wilms tumor 1 (Wt1) is an essential factor for urogenital system development. Teleosts have two wt1s, named as wt1a and wt1b. In this study, the expression pattern of wt1a and wt1b and their functions on the urogenital system were analyzed by in situ hybridization and CRISPR/Cas9. wt1a was found to be expressed in the glomerulus at 3 dah (days after hatching), earlier than wt1b. wan and wt1b were simultaneously expressed in the somatic cells of gonads at 3 dah, while their cell locations were similar, but not identical in adult fish gonads. The wt1a(-/-) fish displayed pericardial edema and yolk sac edema at 3 dah and subsequently expanded as general body edema at 6 dah, failed to develop glomerulus and died during 6-10 dah, whereas the wt1b(-/-) fish were phenotypically normal. Immunohistochemical analyses revealed that the germ cell marker Vasa was expressed, while somatic cell genes Cyp19ala, Amh, Gsdf and Dmrt1 were not expressed in the ipt1a(-/-) gonads at 6 dah. The sex phenotypes of XX and XY in the wtb(-/-) fish were not affected. Real-time PCR revealed that the ovarian cyp19ala expression was up-regulated in XX wt1b(-/-) fish, compared with XX control at 90 dah. Serum estradiol-17 beta level was also up-regulated in XX wt1b(-/-) fish at 90 and 180 dah. The XY wt1b(-/-) fish had normal serum estradiol-17 beta and 11-ketotestosterone levels and remained fertile. These results suggest that Wt1a and Wt1b have different functions in the kidneys and gonads of tilapia.
