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肾上腺皮质束状带特异性表达Cre重组酶转基因小鼠的构建

张宁宁¹, 王长楠¹, 倪鑫^1,2,*

¹海军军医大学生理学教研室，上海 200433；²中南大学湘雅医院分子代谢组学研究中心，长沙 410008

摘要

肾上腺是人体重要的内分泌器官。由于缺乏肾上腺皮质束状带特异性表达Cre酶的工具鼠，目前对肾上腺皮质束状 带细胞中特异表达基因的功能缺乏深入的解析。CYP11B1基因编码类固醇11β-羟化酶，该酶是糖皮质激素合成的关键酶，在 肾上腺皮质束状带中特异性表达。本研究旨在利用CYP11B1基因在束状带特异性表达的特点，构建在肾上腺皮质束状带中 特异性表达Cre重组酶的转基因动物。采用CRISPR/Cas9技术在CYP11B1基因终止密码子位点定点敲入2A-GfpCre表达框，获 得CYP11B1-2A-GfpCre同源重组载体，进而构建CYP11B1Cre小鼠，并通过mTmG和LacZ染色确定Cre酶主要表达在小鼠肾上 腺皮质束状带。在此基础上，本研究还用该工具鼠与胱硫醚-γ-裂解酶(cystathionine γ-lyase, CTH)条件性敲除鼠交配，获得了 肾上腺皮质束状带CTH特异性敲除的小鼠，并证实了该动物肾上腺皮质束状带中CTH表达缺失。以上结果充分说明肾上腺 皮质束状带特异性表达Cre重组酶小鼠构建成功。该工具鼠的成功构建，为深入研究肾上腺皮质束状带相关功能提供了有力工具。

关键词： 肾上腺皮质细胞; 束状带; Cre重组酶; 转基因小鼠

分类号：Q81

Construction of transgenic mice with specific Cre recombinase expression in the zona fasciculata in adrenal cortex

ZHANG Ning-Ning¹, WANG Chang-Nan¹, NI Xin^1,2,*

¹Departmentof Physiology, Navy Medical University, Shanghai 200433, China；²Research Centre for Molecular Metabolomics, Xiangya Hospital, Central Southern University, Changsha 410008, China

Abstract

The adrenal gland is an important endocrine organ of human body. CYP11B1 gene was specifically expressed in the zona fasciculata in adrenal cortex. In order to better study the function of genes specifically expressed in the zona fasciculata in adrenal cortex, the mice with Cre recombinase specifically expressed in the zona fasciculata in adrenal cortex were constructed. It was then confirmed that CYP11B1 was specifically expressed in adrenal glands. Then, using CRISPR/Cas9 technique, CYP11B1-2A-GfpCre recombinant vector was constructed and subsequently injected into the fertilized eggs of mice. It was confirmed that the Cre gene was mainly expressed in the zona fasciculata in adrenal cortex of CYP11B1Cre mice by using mTmG and LacZ staining. The CYP11B1Cre mice were then mated with cystathionine γ-lyase (CTH)f/f mice, thereby generating CTHf/f/CYP11B1Cre mice. It was also confirmed that CTH gene in the zona fasciculata in adrenal cortex was specifically knocked out in these mice. These results suggest that transgenic mice with specific Cre recombinase expression in the zona fasciculata in adrenal cortex were constructed successfully. This animal model can be a powerful tool for the study of the function of genes expressed in the zona fasciculata in adrenal cortex.

Key words: adrenocortical cells; zona fasciculata; Cre recombinase; transgenic mice

收稿日期：2020-01-20　　录用日期：2020-03-20

通讯作者：倪鑫　　E-mail: nixin@smmu.edu.cn

DOI: 10.13294/j.aps.2020.0024

引用本文：

张宁宁, 王长楠, 倪鑫. 肾上腺皮质束状带特异性表达Cre重组酶转基因小鼠的构建[J]. 生理学报 2020; 72 (2): 148-156.

ZHANG Ning-Ning, WANG Chang-Nan, NI Xin. Construction of transgenic mice with specific Cre recombinase expression in the zona fasciculata in adrenal cortex. Acta Physiol Sin 2020; 72 (2): 148-156 (in Chinese with English abstract).