基于转录组学和代谢组学联合分析黑线姬鼠睾丸下降的功能与机制

Functions and mechanisms of testicular descent in Apodemus agrarius based on transcriptomics and metabolomics

摘要:

为了探究黑线姬鼠睾丸下降的功能与机制，分析其基因及代谢物水平的变化规律。本研究使用高通量测序技术和超高效液相色谱技术对黑线姬鼠下降期和正常期睾丸分别进行了转录组学和代谢组学分析。转录组学中基因本体数据库(gene ontology, GO)富集分析得到240个差异基因，包括Spesp1、Izumo1、Hyal5和Fabp9等，京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes, KEGG)富集分析得到52个差异表达基因，包括Pcyt1、Pla2g4e、Gpd1l和Lypla3等。实时荧光定量PCR结果表明，随机选取的6个基因在黑线姬鼠下降期和正常期睾丸中的表达模式与转录组结果一致。代谢组学结果表明，差异代谢集中与睾丸功能相关差异代谢物有28个，包括3-脱氢奎宁酸、α-亚麻酸、磷酸二羟丙酮和1,6-二磷酸果糖等。联合分析结果显示，甘油磷脂代谢、α-亚麻酸代谢和花生四烯酸代谢可能是调控黑线姬鼠睾丸下降及功能发生关键代谢通路。本研究将有助于解读黑线姬鼠睾丸下降对其功能的影响机制，也可为后续深入探究黑线姬鼠种群数量变化机制及实验动物资源开发奠定理论基础。

Abstract:

This study aims to explore the functions and mechanisms of testicular descent in Apodemus agrarius, and analyze the changes in genes and metabolite levels in this process. Illumina NovaSeq and liquid chromatography-mass spectrometry were used for the transcriptomic analysis and metabolomic analysis, respectively, of the normal and descending testis of A. agrarius. Gene ontology (GO) enrichment of the transcriptomic results revealed 240 differentially expressed genes (DEGs), such as Spesp1, Izumo1, Hyal5, and Fabp9. Kyoto encyclopedia of genes and genomes (KEGG) enrichment showed 52 DEGs, including Pcyt1, Pla2g4e, Gpd1l, and Lypla3. The qRT-PCR results were consistent with the transcriptomic results in terms of the expression patterns of six randomly selected genes in the normal and descending testis. The metabolomic results revealed 28 differential metabolites associated with the testicular function, including 3-dehydroquinic acid, α-linolenic acid, dihydroxyacetone phosphate, and fructose 1,6-bisphosphate. The conjoint analysis showcased that glycerophospholipid metabolism, α-linolenic acid metabolism, and arachidonic acid metabolism may be the key metabolic pathways regulating testicular descent in A. agrarius. This study will help to understand the mechanism of testicular descent and lay a theoretical foundation for exploring the mechanisms of the population changes of A. agrarius and developing laboratory animal resources.