摘要
目的利用Cre/LoxP系统建立晶状体组织特异性Msx2基因敲除小鼠模型,并初步研究其表型。方法设计基因敲除方案,获得条件基因敲除小鼠Msx2cko和对照野生型小鼠Msx2wt。通过PCR技术对小鼠或E10.5和E14.5胚鼠进行基因型鉴定,通过整胚LacZ染色鉴定Le-Cre重组酶活性,同时通过对P1小鼠晶状体组织行HE染色进行初步表型观察。结果通过基因型检测证实了Msx2基因敲除小鼠模型建立成功,LacZ染色显示Le-Cre重组酶特异性表达在发育中的小鼠胚胎形成晶状体板的表面外胚叶特定区域。Msx2cko P21小鼠表现为小眼球畸形,眼周至鼻线性脱毛。HE染色显示Msx2cko小鼠晶状体体积减小及晶状体茎形成。结论初步建立了Msx2条件性基因敲除小鼠模型,Msx2条件基因敲除后引起小鼠晶状体发育异常,小眼球畸形。
Objective To establish a Msx2 conditional knockout monsemodel and preliminary explore the phenotypes. Methods The knockout plan was designed and the Msx2cko homozygous mice with complete knockout of Msx2 and wild type control mice were obtained. Genotype type of mice and E10.5-E14.4 embryos were determined by PCR. The expression of Le-Cre from whole mount LacZ staining was visualized. Preliminary phenotype of Msx2eko mice were stained by hematoxylin-eosinand observed. Results The mouse model of Msx2 knockout was determined through genotyping for Msx2 and Cre. LacZ staining showed Le-Cre recombinase was expressed in the specified ectodeml for lens placode forma- tion. The Msx2 conditional deletion causes adult eyes (P21) to be microphthalrnia with missing fur and whiskers around the eye and snout. Hema- toxylin-eosin staining showed small lens size and persistence of the lens stalk. Conclusion The mouse model of conditional knockout of Msx2 is established successfully. The lack of Msx2 in mutant mice results in the abnormality of lens formation and microphthalmia.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2016年第11期964-967,共4页
Journal of China Medical University
基金
国家自然科学基金(81371003)
中国博士后科学基金(2015m570517)
沈阳市科学技术计划(F13-318-1-09)