Comprehensive Multi-Omics Analysis of Muscle Tissue Alterations in Male Induced by Frequent Mating.

During the breeding process of , a male-to-female ratio of 1:3 or higher is typically adopted, so as a result, the quality of the male broodstock significantly influences the quality of the offspring. We observed that overused males exhibited notable changes in body color, particularly in the tail fan region, which turned orange or red due to the excessive accumulation of astaxanthin in the muscles and exoskeleton. Frequent mating also led to a significant decrease in male body weight, with histological analysis revealing disorganized muscle fiber patterns and increased tissue damage. To investigate the molecular mechanisms underlying these physiological changes, we performed transcriptomic and metabolomic analyses of muscle tissues. A total of 1069 differentially expressed genes (DEGs), 540 differentially expressed proteins (DEPs), and 385 differentially expressed metabolites (DEMs) were identified. Pathway analysis revealed that the DEGs were significantly enriched in pathways related to energy metabolism and degenerative diseases, while the DEMs were notably associated with cancer metabolism, signal transduction, substance transport, energy metabolism, nucleic acid metabolism, neurotransmission, immune response, and metabolic diseases. Proteome analysis showed that proteins and lipids were involved in muscle energy supply. These findings suggest that male upregulate energy metabolism in muscles to cope with frequent mating stress, but this adaptation leads to physiological damage. This study provides valuable insights for optimizing male broodstock selection and mating frequency in breeding practices.