条件敲除Sdr9c 7基因小鼠的构建及表型研究

Construction of Sdr9c7 Gene Conditional Knockout Mice and Phenotypic Analysis

利用在Sdr9c7基因第2外显子两侧的内含子区域重组添加有loxP位点的小鼠与上皮细胞特异性表达Cre重组酶的K14Cre小鼠交配产生的子代再进行杂交,获得基因型为Sdr9c7^(fl/fl)K14Cre^(+)的小鼠(上皮细胞Sdr9c7基因特异性敲除小鼠)。在体式镜下观察小鼠表型,用显微镜观察皮肤组织HE染色石蜡切片,拍照观察小鼠皮肤的组织形态结构,利用甲苯胺蓝渗透法检测小鼠皮肤的屏障功能。结果发现:Sdr9c7^(fl/fl)K14Cre^(+)小鼠皮肤发绀,在出生后6~8 h内死亡;HE染色结果表明Sdr9c7^(fl/fl)K14Cre^(+)基因型小鼠的皮肤角质层结构异常,堆积紧密。甲苯胺蓝染液渗透实验中Sdr9c7^(fl/fl)K14Cre^(+)基因型小鼠的皮肤可被染液透过而染成深色,表明其表皮屏障功能不全。

The mice carrying mutant Sdr9c7 gene with loxP site in the intron regions on both sides of its exon 2 were mated with K14-Cre mice,and their offspring were hybridized to obtain epithelial cell Sdr9c7 gene specific knockout mice.The offspring genotyped with Sdr9c7^(fl/fl)K14Cre^(+)were regarded as Sdr9c7 gene epithelial cell conditional knockout mice.Phenotype of mice was observed under stereoscope,HE stained skin paraffin sections were prepared for microscopic observation of the morphological structure of mouse skin,and the barrier function of mouse skin was detected by toluidine blue stain permeation assay.The results revealed that Sdr9c7^(fl/fl)K14Cre^(+)mice exhibited skin cyanosis and died within 6 to 8 hours after birth.HE staining revealed Sdr9c7^(fl/fl)K14Cre^(+)mice had abnormal skin stratum corneum structure which was tightly accumulated.Toluidine blue stain permeation assay showed that the stain could pass through the skin barrier of Sdr9c7^(fl/fl)K14Cre^(+)mice and stain the skin darker,indicating that the epidermal barrier function was disrupted.