雌激素受体Esr1诱导红耳龟早期卵巢分化  

Estrogen receptor Esr1 induces early ovarian differentiation in Trachemys scripta

作　　者：郭金洪 杨洪哲 时翔 胡颖 王圣云 汪思凡 孙伟[1,2] 葛楚天[1,2] GUO Jinhong;YANG Hongzhe;SHI Xiang;HU Ying;WANG Shengyun;WANG Sifan;SUN Wei;GE Chutian(Institute of Animal Sex and Development,Zhejiang Wanli University,Ningbo 315100,Zhejiang,China;College of Biological and Environmental Sciences,Zhejiang Wanli University,Ningbo 315100,Zhejiang,China;College of Marine Life Sciences,Ocean University of China,Qingdao 266000,Shandong,China)

机构地区：[1]浙江万里学院动物性别与发育重点研究所,浙江宁波315100 [2]浙江万里学院生物与环境学院,浙江宁波315100 [3]中国海洋大学海洋生命学院,山东青岛266000

出　　处：《生物工程学报》2024年第7期2308-2321,共14页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(32325049,U22A20529);宁波市自然科学基金(2022J193)。

摘　　要：为探究3种雌激素受体(estrogen receptor) Esr1、Esr2和Gper1在红耳龟早期胚胎性腺分化中的作用,本研究在分析受体基因表达特征的基础上,通过向性腺分化启动前的产雄温度(male-producing temperature,MPT)龟胚分别注射Esr1、Esr2和Gper1激动剂PPT、WAY 200070和G-1,从性腺形态结构、生殖细胞分布模式、性别分化关键基因和蛋白表达分布方面对处理后的胚胎性腺进行了性逆转分析。表达分析结果显示,esr1在性别分化关键时期性腺中的表达量显著高于esr2和gper1 (表达极低),且呈现产雌温度(female-producing temperature,FPT)性腺高表达。功能验证实验显示,PPT处理后的MPT性腺形态结构明显雌性化,生殖细胞呈现雌性分布模式;雄性分化关键基因dmrt1、amh和sox9 m RNA表达明显下降,雌性分化关键基因foxl2和cyp19a1 m RNA表达则显著上升;Amh和Sox9蛋白表达的荧光信号几乎消失,Foxl2和Arom蛋白被激活出现大量表达,表明性腺由雄性逆转为雌性(性逆转率:70.27%)。而WAY 200070和G-1处理后的MPT性腺仍分化为睾丸,雌雄基因和蛋白的表达及分布与雄性性腺类似。结果表明在红耳龟中,单独激活Esr1能够充分启动早期性腺的雌性分化过程,提示雌激素可能通过其受体1 (Esr1)诱导早期卵巢分化。本研究为进一步解析雌激素在龟性别决定和分化中的调控机制提供了参考。This study aims to explore the roles of three estrogen receptors(Esr1,Esr2,and Gper1)in early differentiation of embryonic gonads of Trachemys scripta.The expression characteristics of the receptor genes were studied first.The Esr1,Esr2,and Gper1 agonists PPT,WAY 200070,and G-1 were respectively injected into the embryos at the male-producing temperature(MPT)before initiation of gonadal differentiation.The sex reversal of the treated embryonic gonads was analyzed in terms of morphological structure of gonads,distribution pattern of germ cells,and expression of key genes and proteins involved in sex differentiation.The expression level of esr1 during the critical stage of sex differentiation was higher than those of esr2 and gper1(very low expression)and was particularly high in the gonads at the female-producing temperature(FPT).After treatment with PPT,the MPT gonads presented obviously feminized morphology and structure,with the germ cells exhibiting a female distribution pattern.Furthermore,the mRNA expression levels of the key genes(dmrt1,amh,and sox9)for male differentiation were down-regulated significantly,while those of the key genes(foxl2 and cyp19a1)for female differentiation were up-regulated observably.The fluorescent signals of Amh and Sox9 expression almost disappeared,while Foxl2 and Arom were activated to express abundantly,which fully demonstrated the sex reversal of the gonads from male to female(sex reversal rate:70.27%).However,the MPT gonads treated with WAY 200070 and G-1 still differentiated into testes,and the expression patterns of the key genes and proteins were similar to those in male gonads.The above results demonstrate that activation of Esr1 alone can fully initiate the early female differentiation process of gonads,suggesting that estrogen may induce early ovarian differentiation via Esr1 in Trachemys scripta.The findings provide a basis for further revealing the mechanisms of estrogen regulation in sex determination and differentiation of turtles.

关 键 词：雌激素受体 性腺分化 雌激素 性逆转 红耳龟 

分 类 号：Q953[生物学—动物学]