Lin Jian, Bao Ze Kun, Zhang Qiang, Hu Wei Wei, Yu Qing Hua, Yang Qian
College of Veterinary, Nanjing Agricultural University, Weigang 1, Jiangsu, PR China.
Key Lab of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing Agricultural University, PR China.
Gene. 2015 Jul 10;565(2):228-34. doi: 10.1016/j.gene.2015.04.017. Epub 2015 Apr 10.
Mammary glands are organs for milk production in female mammals. Growth hormone (GH) is known to affect the growth and development of the mammary gland, as well as to increase milk production in dairy goats. This study performed a comprehensive expression profiling of genes expressed in the mammary gland of early involution GH transgenic (n=4) and non-transgenic goats (n=4) by RNA sequencing. RNA was extracted from mammary gland tissues collected at day 3 of involution. Gene expression analysis was conducted by Illumina RNA sequencing and sequence reads were assembled and analyzed using TopHat. FPKM (fragments per kilobase of exon per million) values were analyzed for differentially expressed genes using the Cufflinks package. Gene ontology analysis of differentially expressed genes was categorized using agriGO, while KEGG pathway analysis was performed with the online KEGG automatic annotation server. Our results revealed that 75% of NCBI goat annotated genes were expressed during early involution. A total of 18,323 genes were expressed during early involution in GH transgenic goats, compared with 18,196 expressed genes during early involution of non-transgenic goats. In these expressed genes, the majority (17,589) were ubiquitously expressed in GH transgenic and non-transgenic goats. However, there were 745 differentially expressed genes, 421 of which were upregulated and 324 were downregulated in GH transgenic goats. GO and KEGG pathway analysis showed that these genes were involved in mammary gland physiology, including cell adhesion molecules, ECM-receptor interaction, Jak-STAT signaling pathway, and fat metabolism. Our results demonstrated that the GH receptor was strongly affected in GH transgenic goats, which may activate the IGF-1/Stat3 signaling pathway. Overall, our study provided a global view of the transcriptome during involution of GH transgenic and non-transgenic goats, which increases our understanding of the biology of involution in the goat.
乳腺是雌性哺乳动物的产乳器官。已知生长激素(GH)会影响乳腺的生长发育,还能提高奶山羊的产奶量。本研究通过RNA测序对早期退化阶段的GH转基因山羊(n = 4)和非转基因山羊(n = 4)乳腺中表达的基因进行了全面的表达谱分析。在退化第3天收集乳腺组织并提取RNA。通过Illumina RNA测序进行基因表达分析,并使用TopHat对序列读数进行组装和分析。使用Cufflinks软件包分析差异表达基因的每百万外显子千碱基片段数(FPKM)值。使用agriGO对差异表达基因进行基因本体分析,同时使用在线KEGG自动注释服务器进行KEGG通路分析。我们的结果显示,75%的NCBI注释山羊基因在早期退化阶段表达。GH转基因山羊在早期退化阶段共表达了18323个基因,相比之下,非转基因山羊在早期退化阶段表达了18196个基因。在这些表达的基因中,大多数(17589个)在GH转基因山羊和非转基因山羊中普遍表达。然而,有745个差异表达基因,其中421个在GH转基因山羊中上调,324个下调。GO和KEGG通路分析表明,这些基因参与乳腺生理过程,包括细胞粘附分子、细胞外基质-受体相互作用、Jak-STAT信号通路和脂肪代谢。我们的结果表明,GH转基因山羊中的GH受体受到强烈影响,这可能激活IGF-1/Stat3信号通路。总体而言,我们的研究提供了GH转基因山羊和非转基因山羊退化过程中转录组的全局视图,增进了我们对山羊退化生物学的理解。