Dong Yaqun, Luo Haoyun, Huang Lihua, Liu Lin
Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, China.
Int J Mol Sci. 2025 Jun 10;26(12):5564. doi: 10.3390/ijms26125564.
The , a Lepidopteran pest known for its high fecundity, undergoes a complete metamorphosis, including a distinctive process during which the male testes fuse from two separate organs into a single entity, significantly enhancing its fertility. To elucidate the molecular mechanisms underlying this testicular fusion, this study employed an integrated multi-omics approach to investigate concurrent changes at the transcriptomic and proteomic levels. We identified a series of synchronized alterations on the peritestic larval membrane, including heme binding, peptidase activity, hydrolase activity, metal ion transport, redox reactions, and chitin metabolism, all of which are substantially enriched at specific temporal points during testicular fusion. Nine genes/proteins co-expressed at the mRNA and protein levels were selected for targeted quantitative proteomics (PRM) and quantitative PCR (qPCR) validation, leading to the identification of five genes potentially involved in the testicular fusion process: , , , , and . Notably, the gene , once knocked out, not only disrupted the normal fusion process but also resulted in reduced testis size, thickened peritestic membranes, and abnormal sperm development. Transcriptomic sequencing of the knockout mutant revealed its primary influence on the fusion process by affecting the assembly of the microtubule system and cytoskeleton. This research, for the first time, provides a multi-omics perspective on the response of key signaling pathways and molecular changes during the testicular fusion of and validates the role of the previously uncharacterized gene in this process, offering valuable insights into the complex mechanisms of testicular fusion in this species.
[该鳞翅目害虫]以其高繁殖力而闻名，经历完全变态，包括一个独特的过程，在此过程中雄性睾丸从两个独立的器官融合成一个单一实体，显著提高其生育能力。为了阐明这种睾丸融合背后的分子机制，本研究采用综合多组学方法来研究转录组和蛋白质组水平上的同步变化。我们在睾丸周围幼虫膜上发现了一系列同步变化，包括血红素结合、肽酶活性、水解酶活性、金属离子转运、氧化还原反应和几丁质代谢，所有这些在睾丸融合过程中的特定时间点都大量富集。选择了在mRNA和蛋白质水平上共表达的9个基因/蛋白质进行靶向定量蛋白质组学（PRM）和定量PCR（qPCR）验证，从而鉴定出5个可能参与睾丸融合过程的基因：[基因名称1]、[基因名称2]、[基因名称3]、[基因名称4]和[基因名称5]。值得注意的是，[基因名称5]基因一旦被敲除，不仅会破坏正常的融合过程，还会导致睾丸尺寸减小、睾丸周围膜增厚以及精子发育异常。[基因名称5]基因敲除突变体的转录组测序揭示了其通过影响微管系统和细胞骨架的组装对融合过程的主要影响。本研究首次提供了关于[该物种名称]睾丸融合过程中关键信号通路反应和分子变化的多组学观点，并验证了先前未表征的[基因名称5]基因在这一过程中的作用，为该物种睾丸融合的复杂机制提供了有价值的见解。
