| 张进,刘亚杰,翁敏杰,佘强.2012.Tbx18-Cre基因敲入小鼠的繁殖、鉴定及其应用.动物学杂志,47(3):28-34. |
| 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 |
| 投稿时间：2011-11-25 修订日期：2012-02-29 |
| DOI： |
| 中文关键词: 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 |
| 英文关键词:T-box transcription factor Tbx18 Gene knock-in Cre recombination enzyme gene Polymerase chain reaction (PCR) Heart development |
| 基金项目: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) |
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| 中文摘要: |
| 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Rosa EYFP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。 |
| 英文摘要: |
| To investigate the breeding and genotyping for Tbx18 knock-out mice (Tbx18:Cre knock-in Mus musculus) and the application of Tbx18 knock-out and genetic tracing mice, we introduced into Tbx18: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 Tbx18-expressing epicardial cells within Tbx18:Cre/Rosa26REYFP embryos by cryostat sections. The results indicated that the genotyping of heterozygote offspring, Tbx18 knock-out and Tbx18:Cre/Rosa26REYFP embryos were consistent with Mendel's law of segregation. We also found that there was malformation of the SAN head in Tbx18 knock-out heart compared with the wild-type. The Tbx18-expressing epicardial cells were important cardiac progenitor resource in mouse heart development. These results suggest that Tbx18 knock-out mice is an ideal model for studying the mechanisms of congenital heart diseases. The Tbx18-expressing epicardial cells are candidate cardiac progenitor for cardiac repair and regeneration in heart disease. |
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