Two regions within the proximal steroidogenic factor 1 promoter drive somatic cell-specific activity in developing gonads of the female mouse.
Two regions within the proximal steroidogenic factor 1 promoter drive somatic cell-specific activity in developing gonads of the female mouse.
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近端类固醇生成因子 1 启动子内的两个区域在雌性小鼠性腺发育过程中驱动体细胞特异性活性。
DOI：
10.1095/biolreprod.110.084590
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发表时间：
2011
影响因子：
3.6
通讯作者：
Jorgensen,JoanS
中科院分区：
文献类型：
--
作者：
Gao,Liying;Kim,Youngha;Kim,Bongki;Lofgren,StaceyM;Schultz-Norton,JenniferR;Nardulli,AnnM;Heckert,LeslieL;Jorgensen,JoanS
类固醇生成因子1（SF 1;也称为NR 5A 1和AD 4 BP）的靶标已在下丘脑-垂体-性腺和肾上腺轴的每个水平的细胞内被鉴定，揭示SF 1是主要内分泌系统的主要调节剂。小鼠胚胎在生殖嵴中表达SF 1，直至胚胎第13.5天（E13.5）。此后，表达在雄性中持续存在，在雌性性腺中显著降低，直至出生。我们推测Sf 1在性腺发育过程中的性二态表达是由其启动子的性别特异性调控介导的。为了研究胎儿性腺内的双态调节，我们开发了一种使用E13.5性腺外植体培养物瞬时转染的实验策略，并评估了性双态DNA元件的各种Sf 1启动子结构。proximalSf 1启动子正确地将报告活性靶向XY和XX性腺中的SF 1表达细胞。逐步删除的序列从theSf 1启动子揭示了两个地区，影响调节内女性性腺。这两个序列的突变一起没有造成进一步的破坏报告活性，这表明这两个网站可能协同工作，以促进女性体细胞的活动。凝胶迁移率变动分析和胎儿性腺染色质免疫沉淀的结果表明，TCFAP 2结合到近端启动子ofSf 1内的两个女性特异性位点之一。总之，我们表明，在发育中的睾丸和卵巢内进行的瞬时转染实验是一个强大的工具，以揭示theSf 1启动子内的元素，有助于性别特异性表达。
Targets of steroidogenic factor 1 (SF1; also known as NR5A1 and AD4BP) have been identified within cells at every level of the hypothalamic-pituitary-gonadal and -adrenal axes, revealing SF1 to be a master regulator of major endocrine systems. Mouse embryos express SF1 in the genital ridge until Embryonic Day 13.5 (E13.5). Thereafter, expression persists in the male and is substantially lower in the female gonad until birth. We hypothesize that the sexually dimorphic expression ofSf1during gonadogenesis is mediated by sex-specific regulation of its promoter. To investigate dimorphic regulation within the fetal gonad, we developed an experimental strategy using transient transfection of E13.5 gonad explant cultures and evaluated variousSf1promoter constructs for sexually dimorphic DNA elements. The proximalSf1promoter correctly targeted reporter activity to SF1-expressing cells in both XY and XX gonads. Stepwise deletion of sequences from theSf1promoter revealed two regions that affected regulation within female gonads. Mutation of both sequences together did not cause further disruption of reporter activity, suggesting the two sites might work in concert to promote activity in female somatic cells. Results from gel mobility shift assays and fetal gonad-chromatin immunoprecipitation showed that TCFAP2 binds to one of the two female-specific sites within the proximal promoter ofSf1. Together, we show that transient transfection experiments performed within developing testes and ovaries are a powerful tool to uncover elements within theSf1promoter that contribute to sex-specific expression.