Yes-associated protein and WW-containing transcription regulator 1 regulate the expression of sex-determining genes in Sertoli cells, but their inactivation does not cause sex reversal
Yes-associated protein and WW-containing transcription regulator 1 regulate the expression of sex-determining genes in Sertoli cells, but their inactivation does not cause sex reversal
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DOI：
10.1093/biolre/iox057
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发表时间：
2017-07-01
影响因子：
3.6
通讯作者：
Boyer, Alexandre
中科院分区：
文献类型：
--
作者：
Levasseur, Adrien;Paquet, Marilene;Boyer, Alexandre
Yes相关蛋白（雅普）和WW-containing transcription regulator 1（WWTR 1）是两种功能冗余的转录调节因子，它们是Hippo信号通路的下游效应子，是细胞生长和分化的主要调节因子。为了阐明它们在支持细胞中的作用，用表达Cre重组酶的腺病毒感染来自雅普（flox/flox）; Wwtr 1（flox/flox）动物的原代支持细胞培养物。伴随的雅普和Wwtr 1的失活导致雄性性别分化基因Dhh、Dmrt 1、Sox 9和Wt 1的mRNA水平降低，而诱导参与雌性分化的基因（Wnt 4、Rspo 1和Foxl 2）的mRNA水平降低。SOX 9、FOXL 2和WNT 4蛋白以与它们的mRNA相同的方式响应于雅普和WWTR 1的缺失而受到调节。为了进一步表征雅普和WWTR 1在支持细胞中的作用，我们产生了小鼠模型（雅普（flox/flox）; Wwtr 1（flox/flox）; Amh（cre/+）），其中雅普和Wwtr 1在支持细胞中被条件性缺失。在4 dpp突变小鼠的生精小管中观察到凋亡细胞数量增加，导致成年动物睾丸重量减少和支持细胞数量减少。对4 dpp雅普（flox/flox）; Wwt 1（flox/flox）; Amh（cre/+）小鼠睾丸的基因表达分析表明，支持细胞分化最初发生改变，因为Dhh、Dmrt 1和Sox 9 mRNA水平下调，而Wnt 4 mRNA水平增加。然而，这些基因的表达在老年动物中没有改变。总之，这些结果表明了Hippo信号通路在性别分化机制中的一个新作用。
Yes-associated protein (YAP) and WW-containing transcription regulator 1 (WWTR1) are two functionally redundant transcriptional regulators that are downstream effectors of the Hippo signaling pathway, and that act as major regulators of cell growth and differentiation. To elucidate their role in Sertoli cells, primary Sertoli cell culture from Yap(flox/flox); Wwtr1(flox/flox) animals were infected with a Cre recombinase-expressing adenovirus. Concomitant inactivation of Yap and Wwtr1 resulted in a decrease in the mRNA levels of the male sex differentiation genes Dhh, Dmrt1, Sox9, and Wt1, whereas those of genes involved in female differentiation (Wnt4, Rspo1, and Foxl2) were induced. SOX9, FOXL2, and WNT4 proteins were regulated in the same manner as their mRNAs in response to loss of YAP and WWTR1. To further characterize the role of YAP and WWTR1 in Sertoli cells, we generated a mouse model (Yap(flox/flox); Wwtr1(flox/flox); Amh(cre/+)) in which Yap and Wwtr1 were conditionally deleted in Sertoli cells. An increase in the number of apoptotic cells was observed in the seminiferous tubules of 4 dpp mutant mice, leading to a reduction in testis weights and a decrease in the number of Sertoli cells in adult animals. Gene expression analyses of testes from 4 dpp Yap(flox/flox); Wwtr1(flox/flox); Amh(cre/+) mice showed that Sertoli cell differentiation is initially altered, as Dhh, Dmrt1, and Sox9 mRNA levels were downregulated, whereas Wnt4 mRNA levels were increased. However, expression of these genes was not changed in older animals. Together, these results suggest a novel role of the Hippo signaling pathway in the mechanisms of sex differentiation.Summary SentenceYAP and WWTR1 are required for the maintenance of Sertoli cell differentiation and the suppression of the female sex determination pathway.