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本研究以金黄仓鼠(Mesocricetus auratus)为实验动物，对乳脂球和乳腺进行转录组测序(transcriptome sequencing, RNA-seq),研究其基因表达模式的差异和相关性，探讨应用可多次取样的乳脂球作为研究泌乳特征相关基因表达模式的可行性。采用Trizol法提取金黄仓鼠乳腺组织及乳脂球中的总RNA进行转录组测序，并对乳腺组织和乳脂球中的基因表达模式进行对比分析。结果显示，在金黄仓鼠mRNA中共鉴定出了21 360个基因，其中12 624(59%)个基因在乳脂球和乳腺组织共表达，包括7 908个非差异表达基因(non-differentially expressed genes, NDEGs)和3 992个差异表达基因(differentially expressed genes, DEGs)。NDEGs仅占基因总数的37%,说明乳腺组织和乳脂球中的基因表达模式存在较大差异。对NDEGs和DEGs分别进行GO和KEGG富集分析，结果显示NDEGs和DEGs均主要富集在脂质代谢等泌乳功能相关的信号通路。随机选取4个与泌乳性状相关的NDEGs和DEGs通过实时荧光定量PCR分析其表达量，结果显示其表达趋势与RNA-seq相符。综上所述，乳脂球和乳腺组织中的基因表达模式具有较大差异，乳脂球不适宜完全替代乳腺组织作为基因表达相关研究的材料，仅可应用于部分泌乳相关基因表达的研究。本研究为应用乳脂球代替乳腺组织研究哺乳动物泌乳性状相关基因的表达及调控机制提供了新的理解和数据参考。
Abstract：Golden hamsters(Mesocricetus auratus) were used as the experimental animals to perform transcriptome sequencing(RNA-seq) on milk fat globules and mammary gland in this study. The differences and correlation in gene expression patterns have been stu-died to explore the feasibility of applying multiple milk fat globules that can be sampled multiple times to study gene expression patterns related to lactation characteristics. Total RNA from mammary gland and milk fat globules from golden hamsters was extracted for transcriptome sequencing by Trizol, and gene expression patterns in mammary gland and milk fat globules were compared. The results showed that a total of 21 360 genes were identified from golden hamsters mRNA, of which 12 624(59%) genes were co-expressed in milk fat globules and mammary gland, including 7 908 non-differentially expressed genes(NDEGs) and 3 992 differentially expressed genes(DEGs). NDEGs accounted for only 37% of the total number of genes, indicating large differences in gene expression patterns between milk fat globules and mammary gland. GO and KEGG enrichment analysis of the NDEGs and DEGs showed that both NDEGs and DEGs were mainly enriched in the signaling pathways related to the lactation function such as lipid metabolism. Four randomly selected NDEGs and DEGs associated with lactation traits were analyzed by real time quantitative PCR and the results showed that their expression trends were consistent with RNA-seq. In conclusion, the gene expression patterns in milk fat globules and mammary gland are quite different. Milk fat globules are not suitable to completely replace the breast tissue as materials for gene expression-related studies, and can only be applied to the study of partial lactation-related gene expression. This study provides a new understanding and data reference for the application of milk fat globules instead of mammary gland to study the expression and the regulation mechanism of genes related to lactation traits in mammals.
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基本信息:
DOI：10.13417/j.gab.043.001575
中图分类号:S813
引用信息:
[1]李玉洁,蒋晗偲,徐晓晛,等.金黄仓鼠乳脂球与乳腺组织转录谱的比较分析[J].基因组学与应用生物学,2024,43(Z1):1575-1585.DOI:10.13417/j.gab.043.001575.
基金信息:
广西科技重大专项项目(AA22068099); 广西自然科学基金项目(2021GXNSFBA196036); 广西大学“大学生创新创业训练计划”资助项目(S202210593337)共同资助
2024-10-25
2024-10-25