基于转录组数据挖掘藏羊立毛肌发生的关键基因

doi:10.11843/j.issn.0366-6964.2021.08.010

摘要/Abstract

摘要： 旨在探究调控藏羊皮肤内立毛肌发生的分子机制，挖掘影响立毛肌发生的关键基因。本研究采集了30个75~110日龄健康情况良好的藏羊胚胎，收集背部皮肤组织，制作石蜡组织切片，利用HE染色观察背部皮肤内不同日龄立毛肌的形态学变化特征，推测出立毛肌发生的关键时期为胚龄75~85天（E75~E85）；分别挑选胚龄大约在75和85天各3个背部皮肤组织样品，分样品提取RNA，利用Illumina Hiseq平台进行转录组测序，对测序数据进行质控、过滤、比对，筛选得到差异表达基因，并对差异基因进行功能注释、富集分析、蛋白网络互作和RT-qPCR验证，挖掘调控立毛肌早期发育的关键候选基因。测序结果共得到1 159个差异表达的基因（P<0.05），其中900个基因上调，259个基因下调。随机选择6个差异表达的基因进行RT-qPCR验证，表达趋势与转录组测序结果一致，表明测序结果可靠。通过对差异基因进行GO功能注释和KEGG富集分析，筛选到与立毛肌发育相关的5个生物学过程条目和5个信号通路共47个非冗余基因（如BAMBI、TNNI3、HOXA9等）。本研究表明，藏羊立毛肌开始发育的关键时期约为E75~E85天，该时期内SPP1、BAMBI、TNNI3、HOXA9、SOX15可能为影响藏羊立毛肌发生的关键候选基因。

关键词: 藏羊, 立毛肌, 转录组, 皮肤

Abstract: This study aimed to explore the molecular mechanism and key genes that affect the development of arrector pili muscle in Tibetan sheep dorsal skin. In this study, the dorsal skin tissues of 30 healthy Tibetan sheep embryos aged 75-110 days(E75-E85) were collected. The morphological changes of the arrector pili muscle on different days in the dorsal skin were observed by paraffin tissue section and hematoxylin-eosin (HE) staining. It was inferred that the key period of the occurrence of arrector pili muscle was E75-E85.RNA was extracted from 6 dorsal skin samples(E75 vs. E85, n=3), respectively, and transcriptome sequencing was carried out on the Illumina Hiseq platform. The sequencing data were qualitatively controlled, filtered and compared, and differentially expressed genes were screened. Functional annotation, enrichment analysis, protein-protein interaction network and RT-qPCR were carried out to explore the potential regulatory genes affecting arrector pili muscle development. A total of 1 159 differentially expressed genes were identified (P<0.05), 900 up-regulated genes and 259 down-regulated genes. Six differentially expressed genes were randomly selected for RT-qPCR verification, and the results were consistent with transcriptome sequencing, which indicated that the sequencing results were reliable. Forty seven non-redundant genes (such as BAMBI, TNNI3 and HOXA9, etc) in 5 biological process items and 5 signaling pathways related to arrector pili muscle development were screened through GO function annotation and KEGG enrichment analysis of differentially expressed genes. This study shows that the E75-E85 is probably the key period for the development of arrector pili muscle in Tibetan sheep skin. During this period, SPP1, BAMBI, TNNI3, HOXA9, and SOX15 may be the key genes regulating morphogenesis of arrector pili muscle in Tibetan sheep skin.

Key words: Tibetan sheep, arrector pili muscle, transcriptome, skin

中图分类号:

S826.83

楚金雨, 李绍梅, 杨戈, 牟春燕. 基于转录组数据挖掘藏羊立毛肌发生的关键基因[J]. 畜牧兽医学报, 2021, 52(8): 2171-2180.

CHU Jinyu, LI Shaomei, YANG Ge, MOU Chunyan. Mining Key Genes of Arrector Pili Muscle Development in Tibetan Sheep Based on Transcriptome Data[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2171-2180.

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