ENU诱变构建Jag1基因mRNA剪接异常眼畸形小鼠模型  

Generation of Eye Malformation Mouse Model of Abnormal Splicing of Jag1 mRNA by ENU Inducement

作　　者：徐佳 魏婉娥 吴秋一 释晓纯 王烁 陈兵[2,3,4] XU Jia;WEI Wan’e;WU Qiuyi;SHI Xiaochun;WANG Shuo;CHEN Bing(Editorial Department of Journal of Yangzhou University,Yangzhou 225009,China;College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co‑Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou 225009,China;Ministry of Education Key Lab for Avian Preventive Medicine/Key Laboratory of Jiangsu Preventive Veterinary Medicine,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学学报编辑部,江苏扬州225009 [2]扬州大学兽医学院,江苏扬州225009 [3]扬州大学江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [4]扬州大学教育部禽类预防医学重点实验室/江苏省动物预防医学重点实验室,江苏扬州225009

出　　处：《河南农业科学》2023年第9期148-155,共8页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金项目(31372269,31000987);江苏高校优势学科建设工程资助项目;教育部禽类预防医学点实验室/江苏省动物预防医学重点实验室开放课题(YF202102)。

摘　　要：为建立人类和动物眼病的小鼠模型,采用乙烷基亚硝基脲(ENU)诱导C57BL/6J(B6)小鼠突变手段获得眼病小鼠,利用苏木精-伊红(HE)染色法及免疫组织化学染色技术分析小鼠角膜病变特点,通过连锁分析及位置候选克隆确定致病基因。结果显示,ENU诱变获得1例眼畸形小鼠,小鼠角膜上皮细胞层出现明显空泡,且分化异常,角膜基质层胶原纤维排列杂乱松散,角膜内皮细胞部分脱落。染色体定位将致病基因定位于小鼠第2号染色体微卫星D2Mit107(距着丝粒65.13 cM)与D2Mit423(距着丝粒73.57 cM)之间。对候选基因齿状基因1(Jag1)测序发现,突变杂合子小鼠1条染色体上Jag1基因第24号内含子3′端AG碱基被GG碱基替代,导致第25号外显子存在7个碱基的缺失;蛋白质预测发现,该突变引起Jag1编码区移码及蛋白质编码提前终止。综合分析可知,Jag1基因突变是引起小鼠眼畸形表型的分子基础。In order to create an N‑ethyl‑N‑nitrosourea(ENU)‑induced eye disease mouse model,C57BL/6J male mice were injected intraperitoneally with ENU,HE and immunohistochemical staining analysis of eye sections were conducted for eye malformed mouse,and linkage and sequence analysis were conducted to determine the mutant gene.HE staining analysis showed vacuoles in the surface of the corneal epithelial cells,collagen fibrils were loose in the corneal stromal layer and some endothelial cells were absent in mutant corneas.Immunohistochemistry showed abnormal corneal epithelium differentiation in mutant corneas.The mutation was mapped to mouse chromosome 2 between markers D2Mit107 and D2Mit423,which were located at 65.13 and 73.57 cM,respectively.The Jag1 gene was finally selected as a candidate gene for this mutation.Automated sequencing revealed that the AG base at 3′end of the intron 24 in the Jag1 gene on one chromosome of the mutant heterozygous mouse was replaced by the GG base,resulting in the deletion of 7 bases in the exon 25.Translation of the mutant allele would produce frameshift and truncated protein.The results showed that the Jag1 gene mutation was associated with the eye malformation phenotype.

关 键 词：小鼠 乙烷基亚硝基脲 眼畸形 基因定位 齿状基因1 

分 类 号：S857.6[农业科学—临床兽医学] R771.1[农业科学—兽医学]