TGF-β2、TGF-βR1和TGF-βR2在小尾寒羊卵泡发育过程中的功能研究  

Functional study on TGF-β2,TGF-βR1 and TGF-βR2 during follicular development in small-tailed Han sheep

作　　者：肖玉宝 李明娜 王继卿 郝志云 任思语 郭雅静 何兆华 刘圆 邵鹏阳 陈占朝 XIAO Yubao;LI Mingna;WANG Jiqing;HAO Zhiyun;REN Siyu;GUO Yajing;HE Zhaohua;LIU Yuan;SHAO Pengyang;CHEN Zhanzhao(College of Animal Science and Technology/Gansu Key Laboratory of Herbivorous Animal Biotechnology,Gansu Agricutural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物科学技术学院甘肃省草食动物生物技术重点实验室,兰州730070

出　　处：《黑龙江畜牧兽医》2024年第23期55-61,126,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31960664,31560634);甘肃农业大学学科团队项目(GAU-XKTD-2022-21);兰州市青年科技人才创新项目(2023-QN-187)。

摘　　要：为了探究转化生长因子-β2(TGF-β2)、转化生长因子-βR1(TGF-βR1)和转化生长因子-βR2(TGF-βR2)在小尾寒羊卵巢、卵泡中的生物学功能,试验对雌性小尾寒羊实施同期发情,采用免疫荧光检测、实时荧光定量PCR扩增、Western-blot等方法检测TGF-β2、TGF-βR1和TGF-βR2在不同生理阶段卵巢中的表达模式;随后分离不同直径卵泡,探究目的蛋白在不同大小卵泡中的表达差异。结果表明:TGF-β2基因及其编码蛋白在撤栓后第18小时时卵巢中的相对表达量显著高于撤栓后第42小时时(P<0.05);TGF-βR1基因及其编码蛋白在撤栓后第18,42小时时卵巢中的相对表达量差异不显著(P>0.05);TGF-βR2基因编码蛋白在撤栓后第18小时时卵巢中的相对表达量显著高于撤栓后第42小时时(P<0.05),而其mRNA相对表达量差异不显著(P>0.05)。TGF-β2、TGF-βR1基因及其编码蛋白在大卵泡中的相对表达量显著高于中卵泡(P<0.05);TGF-βR2基因及其编码蛋白在大卵泡和中卵泡中的相对表达量差异极显著(P<0.01)。TGF-β2蛋白主要表达于卵泡膜细胞中,TGF-βR1蛋白主要表达于颗粒细胞中,TGF-βR2蛋白在颗粒细胞和卵泡膜细胞中均高表达。说明TGF-β2、TGF-βR1和TGF-βR2对小尾寒羊卵泡选择至优势化阶段的发育具有调控作用,并且TGF-β2可能还参与发情起始至排卵前卵巢的其他生理过程。In order to explore the biological functions of transforming growth factor-β2(TGF-β2),transforming growth factor-βR1(TGF-βR1)and transforming growth factor-βR2(TGF-βR2)in ovarian follicles of small-tailed Han sheep,female small-tailed Han sheep were subjected to simultaneous estrus,immunofluorescence(IF),real-time quantitative PCR and Western-blot were used to detect the expression patterns of TGF-β2,TCF-βR1 and TGF-βR2 in the ovaries at different physiological stages.Subsequently,different diameter follicles were isolated to explore the difference in the expression of target protein in follicles of different sizes.The results showed that the expression level of TGF-β2 gene and its coding protein in the ovary at 18 h after thrombosis withdrawal was significantly higher than that at 42 h(P<0.05),there was no significant difference in the expression of TGF-βR1 gene and its coding protein in the ovary at 18 h and 42 h after thrombosis withdrawal(P>0.05),and the expression level of TGF-βR2 protein in the ovary at 18 h after thrombosis withdrawal was significantly higher than that at 42 h(P<0.05),but there was no significant difference in the mRNA level(P>0.05).The relative expression levels of TCF-β2 and TCF-βRl genes and their coding proteins in large fllicles were significantly higher than those in middle follicles(P<0.05),and there were significant dfferences in the relative expression level of TCF-βR2 gene and its coding protein between large follicles and middle folicles(P<0.01).TGF-β2 protein was mainly expressed in follicular membrane cells;TCF-βR1 protein was mainly expressed in granulosa cells,and TGF-βR2 protein was highly expressed in granulosa cells and follicular membrane cells.These results indicated that TGF-β2,TGF-βR1 and TCF-βR2 had a regulatory effect on the development of follicles from follicle selection to dominance in small-tailed Han sheep,and TGF-β2 might also be involved in other physiological processes in the ovaries from estrus initiation to pre-ovulation.

关 键 词：TGF-Β2 TGF-ΒR1 TGF-βR2 小尾寒羊 卵泡 

分 类 号：S826[农业科学—畜牧学]