Regulation of Female Folliculogenesis by Tsp1a in Nile Tilapia (Oreochromis niloticus)
Regulation of Female Folliculogenesis by Tsp1a in Nile Tilapia (Oreochromis niloticus)
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tsp1a 对尼罗罗非鱼 (Oreochromis niloticus) 雌性卵泡发生的调节
DOI：
10.3390/ijms21165893
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发表时间：
2020-08-01
影响因子：
5.6
通讯作者：
Wang, Deshou
中科院分区：
文献类型：
--
作者：
Jie, Mimi;Ma, He;Wang, Deshou
据报道，TSP1参与了多种生物学过程，包括激活转化生长因子-β信号通路，调节创伤修复和肿瘤生长过程中的血管生成，但其在卵巢卵泡发生中的作用尚不清楚。在本研究中，免疫荧光法发现Tsp1a在尼罗罗非鱼卵巢I期至IV期卵泡的卵原细胞和颗粒细胞中有表达，经CRISPR/Cas9获得纯合突变体。Tsp1a基因突变导致卵原细胞增多，次级生长卵泡减少，卵巢发育迟缓。突变型卵巢卵原细胞中细胞增殖标记物增殖细胞核抗原的表达显著上调。此外，转录转录分析显示，DNA复制相关基因的表达显著上调，而抑制细胞增殖和促进细胞分化的cAMP和MAPK信号通路基因的表达显著下调。此外，突变体中芳香化酶(Cyp19a1a)的表达和血清17β-雌二醇(E_2)浓度显著降低。这些结果表明，缺乏tsp1α可促进卵原细胞的增殖，抑制卵原细胞的分化，进而导致卵原细胞增多，次级生长卵泡减少，E2水平降低。综上所述，我们的结果表明attsp1a在尼罗罗非鱼卵泡发生中是必需的。
TSP1 was reported to be involved in multiple biological processes including the activation of TGF-beta signaling pathways and the regulation of angiogenesis during wound repair and tumor growth, while its role in ovarian folliculogenesis remains to be elucidated. In the present study, Tsp1a was found to be expressed in the oogonia and granulosa cells of phase I to phase IV follicles in the ovaries of Nile tilapia by immunofluorescence.tsp1ahomozygous mutants were generated by CRISPR/Cas9. Mutation oftsp1aresulted in increased oogonia, reduced secondary growth follicles and delayed ovary development. Expression of the cell proliferation marker PCNA was significantly up-regulated in the oogonia of the mutant ovaries. Furthermore, transcriptomic analysis revealed that expressions of DNA replication related genes were significantly up-regulated, while cAMP and MAPK signaling pathway genes which inhibit cell proliferation and promote cell differentiation were significantly down-regulated. In addition, aromatase (Cyp19a1a) expression and serum 17 beta-estradiol (E2) concentration were significantly decreased in the mutants. These results indicated that lackingtsp1aresulted in increased proliferation and inhibited differentiation of oogonia, which in turn, resulted in increased oogonia, reduced secondary growth follicles and decreased E2. Taken together, our results indicated thattsp1awas essential for ovarian folliculogenesis in Nile tilapia.