Bulk RNA-seq analysis of developing ocular anterior segments from wildtype and Col4a1 mutant mice

COL4A1 mutations cause Gould syndrome, a multi-system disorder which is characterized by a broad range of cerebrovascular, ocular, muscular, renal, or cardiac manifestations. To investigate the patheogenic mechanism of Col4a1 mutations in ocular anterior segment dysgenesis, we used a Col4a1 mutant mouse model that recapitulate many aspects of the pathophysiological hallmarks of Gould syndrome including anterior segment dysgenesis. We then isolated developing anterior segments from wildtype and mutant mice and performed differntial expression analysis using bulk RNAseq. Anterior segments (cornea, iris, ciliary body and ocular drainage structures) were dissected from P0 Col4a1+/+ (wildtype) and Col4a1+/G1344D (mutant) eyes and comparative gene expression profiling analysis were performed.COL4A1基因突变（COL4A1 mutation）可引发古尔德综合征（Gould syndrome），这是一类多系统紊乱疾病，其特征为广泛的脑血管、眼部、肌肉、肾脏及心脏受累表现。为探究Col4a1基因突变在眼前节发育异常中的致病机制，我们使用了一款可重现古尔德综合征多种病理生理标志性特征（包括眼前节发育异常）的Col4a1突变小鼠模型。随后我们从野生型与突变小鼠体内分离发育中的眼前节组织，并通过批量RNA测序（bulk RNAseq）开展差异表达分析。具体操作如下：从出生后0天（P0）的Col4a1+/+（野生型）及Col4a1+/G1344D（突变型）小鼠眼球中分离眼前节组织（包含角膜、虹膜、睫状体与眼部引流结构），并完成比较基因表达谱分析。