RNA-Seq分析小鼠腭发育关键时期基因的时空表达

doi:10.13701/j.cnki.kqyxyj.2018.12.022

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (12): 1356-1359.DOI: 10.13701/j.cnki.kqyxyj.2018.12.022

RNA-Seq分析小鼠腭发育关键时期基因的时空表达

彭瑶,苏超南^#,王欣欢,王坤,乔玮玮,高倩,孟柳燕^*

武汉大学口腔医学院口腔生物医学工程教育部重点实验室湖北武汉 430079

收稿日期:2018-04-25 出版日期:2018-12-28 发布日期:2018-12-27
通讯作者: 孟柳燕,E-mail:mengliuyan@whu.edu.cn
作者简介:彭瑶(1993~ ),女,湖南衡阳人,研究生在读,主要从事唇腭裂病因研究。苏超南 (1989~ ), 女,河南濮阳人,硕士,主要从事唇腭裂及显微根尖手术方面研究。
基金资助:
国家自然科学基金(编号:81571438)

RNA-Seq Analysis of Temporal and Spatial Expression of Genes in the Key Period of Palatogenesis in Mice.

PENG Yao, SU Chao-nan^#, WANG Xin-huan, WANG Kun, QIAO Wei-wei, GAO Qian, MENG Liu-yan^*

Key Laboratory of Oral Biomedical Engineering of Ministry of Education, School of Stomatology, Wuhan University, Wuhan 430079, China.

Received:2018-04-25 Online:2018-12-28 Published:2018-12-27

摘要/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用

关键词: 腭发育, 唇腭裂, RNA测序

Abstract: Objective: To construct DEGB during palatogenesis in mice, and to explore the key regulatory factors and regulation mechanism of palate development.Methods: The embryonic palatal shelves of pregnant C57BL/6 at Ed13.5, Ed14.5, Ed15.5, and Ed16.5 were collected respectively for RNA-seq to screen DEGs.Selected DEGs were validated and analyzed by qPCR and immunohistochemistry.Results: RNA-seq identified 243 DEGs (27 DEGs were down-regulated and 216 DEGs were up-regulated) at the first stage of palatal development (Ed13.5-Ed14.5), 208 DEGs (30 DEGs down-regulated and 178 DEGs up-regulated) at the second stage (Ed14.5-Ed15.5), and 262 DEGs between E15.5 and Ed16.5 (31 DEGs down-regulated and 231 DEGs up-regulated) at the third stage (Ed15.5-Ed16.5).The expression of DEGs related with micro RNA and crystallin family changed significantly.Immunohistochemistry showed that Spp1 was gradually increased in palatal mesenchyme from Ed13.5 to Ed16.5.Conclusion: Members in micro RNA family and crystallin family and Spp1 gene play a role in the palatogenesis.

Key words: Palate development, Cleft lip and palate, RNA-Seq

彭瑶,苏超南,王欣欢,王坤,乔玮玮,高倩,孟柳燕. RNA-Seq分析小鼠腭发育关键时期基因的时空表达[J]. 口腔医学研究, 2018, 34(12): 1356-1359.

PENG Yao, SU Chao-nan, WANG Xin-huan, WANG Kun, QIAO Wei-wei, GAO Qian, MENG Liu-yan. RNA-Seq Analysis of Temporal and Spatial Expression of Genes in the Key Period of Palatogenesis in Mice.[J]. Journal of Oral Science Research, 2018, 34(12): 1356-1359.

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RNA-Seq分析小鼠腭发育关键时期基因的时空表达

RNA-Seq Analysis of Temporal and Spatial Expression of Genes in the Key Period of Palatogenesis in Mice.

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