POMC神经元Isl1基因条件性敲除小鼠模型构建及表型分析  

Construction and Phenotypic Analysis of Conditional Knockout Mice for the POMC Neuron Isl1 Gene

作　　者：黄笠纹 李雪[1,2] 潘朋歌 刘美辰 张茜 马萍[2] 尹海涛 裴秀英 HUANG Liwen;LI Xue;PAN Pengge;LIU Meichen;ZHANG Qian;MA Ping;YIN Haitao;PEI Xiuying(Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education,Ningxia Medical University,Yinchuan 750004,China;School of Basic Medical Sciences,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学生育力保持教育部重点实验室,银川750004 [2]宁夏医科大学基础医学院,银川750004

出　　处：《宁夏医科大学学报》2023年第6期553-559,共7页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(82160290);宁夏自然科学基金项目(2021AAC03133;2021AAC03134);宁夏医科大学重点项目(XZ2022003;XZ2020003)

摘　　要：目的构建POMC神经元上Isl1基因条件性敲除小鼠模型,并初步分析其表型。方法利用Cre/loxP系统构建条件性基因敲除小鼠模型,通过Isl1flox/+自交获得Isl1flox/flox小鼠,再将Isl1flox/flox小鼠与POMCCre基因型小鼠交配得到POMCCre;Isl1flox/+雄鼠,POMCCre;Isl1flox/+雄鼠再与Isl1flox/flox雌鼠交配,通过PCR进行基因型鉴定,得到POMC神经元上Isl1条件性敲除小鼠(POMCCre;Isl1flox/flox,缩写为Isl1ΔPOMC),免疫印迹法验证其敲除效率。通过功能学及组织病理学方法,观察主要组织器官是否存在自发病变。结果成功构建了Isl1ΔPOMC小鼠,Isl1ΔPOMC小鼠下丘脑弓状核中ISL1敲除效率>90%(P<0.05),其他组织中ISL1表达水平无变化(P>0.05),证明POMC神经元Isl1基因敲除小鼠模型构建成功。Isl1ΔPOMC小鼠呈现青春期后自发性进行性肥胖、代谢综合征及生育能力障碍等表型。结论应用Cre/loxP技术成功构建了POMC神经元Isl1基因条件性敲除小鼠,为研究Isl1在POMC神经元中生物学功能和调控机制提供了重要的动物模型。Objective To establish conditional knockout mice for Isl1 on POMC neurons and preliminary analysis of their phenotype.Methods A conditional knockout mouse model was constructed using the Cre/LoxP system,and Isl1flox/flox mice were obtained by Isl1flox/+self-crossing,then Isl1flox/flox mice were mated with POMCCre genotype mice to obtain POMCCre;Isl1flox/+male mice,POMCCre;Isl1flox/+male mice were then mated with Isl1flox/flox females and genotyped by PCR to obtain Isl1 conditional knockout mice on POMC neurons(POMCCre;Isl1flox/flox,abbreviated Isl1ΔPOMC),and the knockout efficiency was verified by immunoblotting.The presence of spontaneous lesions in major tissues and organs were investigated by functional and histopathological methods.Results The results showed that the knockout efficiency of ISL1 in the hypothalamic arcuate nucleus of Isl1ΔPOMC mice was greater than 90%(P<0.05),while the expression level of ISL1 in other tissues did not alter(P>0.05),which proved the POMC neuronal Isl1 knockout mouse model was established successfully.The phenotype of Isl1ΔPOMC mice showed spontaneous progressive obesity,metabolic syndrome and impaired fertility after puberty.Conclusion We successfully constructed Isl1 conditional knockout mice in POMC neurons by using Cre/LoxP technology,providing an important animal model for studying the biological function and regulatory mechanism of Isl1 in POMC neurons.

关 键 词：POMC神经元 胰岛素基因增强蛋白1 肥胖 生育力 

分 类 号：R589.2[医药卫生—内分泌]