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- 2018
RNA-Seq分析小鼠腭发育关键时期基因的时空表达
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Abstract:
摘要 目的: 构建小鼠腭发育时期差异表达基因库(differentially expressed genes bank,DEGB),探究腭发育关键调控因子和调控机制。方法: 分别收集C57BL/6孕鼠妊娠13.5、14.5、15.5、16.5 d(embryonic day,Ed)胚胎腭板,RNA测序技术筛查差异表达基因(differentially expressed genes,DEGs),选择部分基因进行功能注释,定量聚合酶链反应( quantitative polymerase chain reaction,qPCR )验证和免疫组织化学染色。结果: 腭发育第一阶段(Ed13.5~Ed14.5)筛出243个DEGs(27个下调,216个上调);第二阶段(Ed14.5~Ed15.5)筛出208个DEGs(30个下调,178个上调);第三阶段(Ed15.5~Ed16.5)筛出262个DEGs(31个下调,231个上调)。其中微小RNA(micro RNA,miRNA)和晶体蛋白家族多个成员表达变化显著。免疫组织化学结果显示Ed13.5~Ed16.5腭间充质中分泌型磷酸蛋白1(secreted phosphoprotein,Spp1)表达量逐渐升高。结论: miRNA家族、晶体蛋白家族成员以及Spp1基因在腭形成时期可能发挥关键作
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