模拟失重对家蚕胚胎发育的影响及基因表达变化的研究

Effects of simulated microgravity on embryonic development and gene expression alterations in Bombyx mori

目的研究模拟失重对家蚕胚胎发育的影响与基因表达变化。方法将通过三维随机回转器模拟失重环境处理的家蚕胚胎作为回转组。对照组是未进行回转处理的同批次家蚕卵。在家蚕胚胎的中期和孵化期分别对回转组和对照组样本进行转录组测序。观察家蚕胚胎发育状态和蚁蚕孵化时间,分析转录组中差异表达的基因。通过GO和KEGG富集分析,探讨差异基因涉及的信号通路,并使用qRT-PCR验证这些差异基因的表达。结果模拟失重使家蚕胚胎发育提前。GO富集分析显示,与对照组相比,回转组胚胎和孵化蚁蚕表达变化的基因均主要富集于催化活性、膜结构和代谢的生物学过程。KEGG通路富集分析显示,与对照组相比,回转组胚胎表达变化的基因主要富集在核糖体和RNA转运通路,而回转组孵化蚁蚕主要富集在代谢、粘着斑和紧密连接通路。qRT-PCR结果表明,与对照组相比,回转组胚胎中的有机阳离子转运蛋白、突触囊泡糖蛋白2B等基因表达显著上调,回转组蚁蚕中的氨肽酶N等基因表达显著上调,而胶原蛋白和钙结合表皮生长因子结构域等基因表达显著下调。结论模拟失重可以加速家蚕胚胎的发育进程,且家蚕胚胎期基因表达对失重环境很敏感。

Objective To investigate the effects of simulated microgravity on embryonic development and gene expression alterations in Bombyx mori.Methods Bombyx mori embryos treated in simulated microgravity with 3D clinostat were used as treatment group.The control group consisted of untreated Bombyx mori embryos from the same batch.Transcriptomic sequencing was conducted on randomly selected samples of Bombyx mori embryos or newly-hatched larvae from both the control and treatment groups.Embryonic development status and larval hatching time of Bombyx mori were observed,and the differentially expressed genes in the transcriptome were analyzed.The signaling pathways involved in differentially expressed genes were investigated through GO and KEGG enrichment analysis,and the expression of these differential genes was validated by qRT-PCR.Results Simulated microgravity advanced the development of Bombyx mori embryos.GO enrichment analysis revealed that,compared with the control group,differentially expressed genes of embryos and newly-hatched larvae in the treatment group were primarily enriched in biological processes related to catalytic activity,membrane structure,and metabolism.KEGG pathway enrichment analysis showed that,compared with the control group,differentially expressed genes of embryos in the treatment group were primarily enriched in ribosome and RNA transport pathways,while those of newly-hatched larvae in the treatment group were mainly enriched in metabolism,focal adhesion,and tight junction pathways.qRT-PCR results indicated that,compared with the control group,embryos in the treatment group exhibited significant upregulation of genes such as organic cation transporter protein and synaptic vesicle glycoprotein 2B,and newly-hatched larvae in the treatment group showed significant upregulation of genes such as aminopeptidase N,and significant downregulation of genes such as collagen and calcium-binding EGF domain.Conclusion Simulated microgravity speeds up Bombyx mori embryonic development,and the gene expression of Bombyx mori is sensitive to microgravity.