摘要
为探究DNA分化抑制因子3(ID3)影响卵泡选择的分子机制,试验观察比较了35周龄与75周龄京红1号蛋鸡卵泡形态、大小及数量差异,采用实时荧光定量PCR检测ID3在不同发育阶段卵泡中的表达,并通过CRISPRi技术与rhBMP2蛋白分别调控鸡小黄卵泡颗粒细胞中ID3的表达,检测卵泡选择相关基因卵泡刺激素受体(FSHR)与类固醇生成急性调节蛋白(StAR)相对表达量。结果显示:35周龄京红1号蛋鸡等级卵泡数目显著高于75周龄蛋鸡(P<0.05);ID3在35周龄京红1号蛋鸡不同发育阶段卵泡中表达差异不显著(P>0.05),但75周龄蛋Fx(F6/7)卵泡中ID3表达显著高于其他卵泡(P<0.05),且75周龄蛋鸡的大白卵泡、小黄卵泡、Fx卵泡、F3卵泡、F1卵泡及卵巢基质中的ID3表达显著高于35周龄蛋鸡(P<0.05);通过CRISPRi抑制ID3表达时,FSHR与StAR的表达会显著增加(P<0.05);10、50、200 ng/mL rhBMP2能显著诱导ID3的表达(P<0.05),FSHR与StAR的表达下降,但差异不显著。研究证实ID3与鸡卵泡发育选择密切相关,并明确其影响卵泡选择的分子机制,对延长蛋鸡产蛋持久性具有重要意义。
To explore the mechanism that inhibitor of DNA binding and/or differentiation 3(ID3)affect follicular selection,this study compared the differences of follicle morphology and number of Jinghong No.1 laying hens at 35 and 75 weeks of age,and detected the ID3 gene expression in follicles at different developmental stages by real-time quantitative PCR.Furthermore,the expression of ID3 gene in granulosa cells of small yellow follicles was perturbed by CRISPRi technology or rhBMP2 protein,and the expression of follicle selection-related genes follicle-stimulating hormone receptor(FSHR)and steoidogenic acute regulatory(StAR)were detected.The results showed that the number of hierarchical follicles in Jinghong No.1 laying hens at 35 weeks of age was significantly higher than that in laying hens at 75 weeks of age(P<0.05).The expression of ID3 gene in Fx follicles was significantly higher than that in other follicles(P<0.05).However,the expression of ID3 was not significantly different in the follicles of different developmental stages of hens at 35 weeks of age(P>0.05),and the ID3 gene expression in large white follicles,small yellow follicles,Fx(F6/7)follicles,F3follicles,F1follicles and ovary of laying hens at 75 weeks of age was significantly higher than that laying hens at 35 weeks of age(P<0.05).When ID3 expression was inhibited by CRISPRi,the expressions of FSHR and StAR were significantly increased(P<0.05);10,50,and 200 ng/mL rhBMP2 could significantly induce the expression of ID3(P<0.05),but the expression regulation of FSHR and StAR was not significantly decreased.It was concluded that ID3 was closely related to follicular selection in chickens,and the molecular mechanism of its impact on follicular selection were clarified,which was of great significance for prolonging egg laying persistence in laying hens.
作者
王丹
王方
张泽春
邹可欣
李艾欣
魏泽辉
WANG Dan;WANG Fang;ZHANG Zechun;ZOU Kexin;LI Aixin;WEI Zehui(College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100)
出处
《中国家禽》
北大核心
2023年第2期1-7,共7页
China Poultry
基金
国家自然科学基金项目(32172736)。
