Ⅰ型毛-肝-肠综合征小鼠模型构建及表型分析  

Phenotypic evaluation of Ttc37 knockout mouse as typeⅠtrichohepato-enteric syndrome model

作　　者：李名亚 王雪琳[1] 韦晔 杨培红 孙磊[1] LI Ming-ya;WANG Xue-lin;WEI Ye;YANG Pei-hong;SUN Lei(Institute of Developmental Biology and Molecular Medicine,School of Life Sciences,Fudan University,Shanghai 200433,China;General Practice of Traditional Chinese Medicine,Pujin Community Health Service Centre,Shanghai 201112,China;School of Life Sciences,Inner Mongolia University,Hohhot 010070,Inner Mongolia Autonomous Region,China)

机构地区：[1]复旦大学发育生物学研究所,生命科学学院,上海200433 [2]浦锦社区卫生服务中心中医全科,上海201112 [3]内蒙古大学生命科学学院,呼和浩特010070

出　　处：《复旦学报（医学版）》2024年第2期249-256,共8页Fudan University Journal of Medical Sciences

摘　　要：目的 建立基于Ttc37(Tetratricopeptide Repeat Domain 37)基因缺失的Ⅰ型毛-肝-肠综合征(trichohepato-enteric syndrome,THES)的小鼠疾病模型。方法 利用CRISPR/CAS9技术在小鼠Ttc37基因中插入loxP序列构建Ttc37flox品系,通过与全身表达的CAG-Cre品系交配产生全身敲除小鼠Ⅰ型THES动物模型(Ttc37flox/flox;CAG-Cre),利用荧光定量PCR和Western blot确认敲除效果。选取8周龄小鼠,对皮肤、脾脏、肝脏、膀胱和胃肠道等主要组织进行苏木精-依红染色和病理分析,对血清中的谷草转氨酶(aspartate aminotransferase,AST)和谷丙转氨酶(alanine aminotransferase,ALT)进行酶学检测,对血清中的血红蛋白进行检测,注射抗原后利用ELISA检测免疫球蛋白IgM和IgG水平。结果 Ttc37flox/flox;CAG-Cre表现出与Ⅰ型THES相似的毛发、皮肤、B细胞和眼睛发育异常表型;常规饲养条件下未表现出肝脏、胃肠道、膀胱和血红蛋白异常。结论 Ⅰ型THES的小鼠疾病模型构建成功,可用于病理机制研究。Objective To establish a mouse model of typeⅠtricho-hepato-enteric syndrome(THES)induced by Ttc37 deficiency.Methods Ttc37 flox strain was established by site-specifically inserted loxP sites into Ttc37 gene via CRISPR/CAS9 technology.Ubiquitously expressed CAG-Cre was introduced for all-tissue removal of Ttc37 in Ttc37flox/flox;CAG-Cre mice.The knock-out effect was confirmed by fluorescence quantitative PCR and Western blot.Phenotypic evaluations were conducted in 8-week-old mice including hematoxylin-eosin staining of skin,spleen,liver,bladder,and gastrointestinal tract(GI),serum enzyme activity assay of aspartate aminotransferase(AST)and alanine aminotransferase(ALT),measurement of serum hemoglobin level,and ELISA for IgG and IgM level upon antigen immunization.Results Similar to typeⅠTHES patients,Ttc37flox/flox;CAG-Cre mice exhibited impaired development of hair shaft,epidermis,B cell and eyes,while liver,GI,bladder and serum hemoglobin level seemed normal under unstressed condition.Conclusion A novel mouse model of typeⅠTHES was constructed successfully,which was applicable for pathological study.

关 键 词：小鼠模型 毛-肝-肠综合征(THES) Ttc37基因 

分 类 号：R596.2[医药卫生—内科学] R-332[医药卫生—临床医学]