Tbx18-Cre基因敲入小鼠的繁殖、鉴定及其应用  

The Breeding,Genotyping and Application of Tbx18-Cre Knock-in Mice

作　　者：张进[1] 刘亚杰[1] 翁敏杰[1] 佘强[1] 

机构地区：[1]重庆医科大学附属第二医院心血管内科,重庆400010

出　　处：《动物学杂志》2012年第3期28-34,共7页Chinese Journal of Zoology

基　　金：国家自然科学基金面上项目(No.30971213);重庆市卫生局重点项目(No.2009-1-13);2010重庆医科大学校级重点项目(No.201010)

摘　　要：为了探讨Tbx18-Cre基因敲入小鼠(Tbx18:Cre knock-in Mus musculus)的繁殖、鉴定及Tbx18基因敲除小鼠和遗传示踪小鼠模型的应用,将Tbx18-Cre基因敲入杂合子小鼠进行繁殖,应用PCR法鉴定其子代基因型。将子代雌雄杂合子小鼠互交,应用H.E染色观察Tbx18基因敲除胚鼠心的形态学变化。将杂合子小鼠与RosaEYFP报告小鼠交配,应用心冰冻切片技术观察Tbx18:Cre/Rosa26REYFP双转基因遗传示踪胚鼠心内Tbx18阳性心外膜祖细胞发育命运。结果表明,用于繁殖、基因敲除研究及基因遗传示踪的子代基因型均符合孟德尔遗传规律。同时心H.E染色和心冰冻切片发现,Tbx18敲除小鼠心窦房结发育存在缺陷,而Tbx18阳性心外膜祖细胞是心发育重要的祖细胞来源。研究结果揭示,Tbx18-Cre基因敲除小鼠是研究先天性心脏病发病机制的理想模式动物,Tbx18阳性心外膜祖细胞可能是心脏病患者心脏修复和再生潜在的种子细胞。To investigate the breeding and genotyping for Tbxl8 knock-out mice （Tbxl8：Cre knock-in Mus musculus） and the application of Tbxl8 knock-out and genetic tracing mice, we introduced into Tbxl8：Cre knock-in heterozygote mice and breed offspring. The genotyping of the offspring were performed by PCR using genomic DNA. We observed the morphology of knock-out （KO） hearts from male and female heterozygate mice inter-crossed by Hematoxylin and eosin-staining（ H. E）analysis. We also observed the fate of TbxlS-expressing epicardial cells within Tbxl8： Cre/Rosa26REYrP embryos by cryostat sections. The results indicated that the genotyping of heterozygote offspring, Tbxl8 knock-out and Tbx18：Cre/Rosa26REYrP embryos were consistent with Mendel＇s law of segregation. We also found that there was malformation of the SAN head in Tbxl8 knock- out heart compared with the wild-type. The Tbxl8-expressing epicardial cells were important cardiac progenitor resource in mouse heart development. These results suggest that Tbxl8 knock-out mice is an ideal model for studying the mechanisms of congenital heart diseases. The Tbxl8-expressing epicardial cells are candidate cardiac progenitor for cardiac repair and regeneration in heart disease.

关 键 词：转录因子Tbx18 基因敲入 Cre重组酶基因 聚合酶链式反应(PCR) 心发育 

分 类 号：R-332[医药卫生]