Abstract
Skin color is a crucial economic trait for commercial fishes. Recently, excluding wild-type grayish black alternations, pufferfish (Takifugu fasciatus) with white and black colors were observed in China. The molecular mechanism underlying these fascinating skin color differences is still unknown. We examined T. fasciatus individuals with three different skin colors, namely, control (C), white (W), and black (B), utilizing histology, proteomics, metabolomics, and Western blotting techniques. Histological observations revealed that melanin content plays a pivotal role in determining skin color. Proteomics analysis comparing control and black individuals revealed significant enrichment of melanin-related proteins, including phosphatidylinositol 3-kinase (Pi3k), solute carrier family 2 (Slc27a), dual specificity phosphatase (Dusp), and Wnt8a. Similarly, compared to controls, white individuals exhibited significant enrichment of Ellis-van-Creveld syndrome protein 1 (Evc), smoothened (Smo). A total of 11 proteins that were linearly correlated with changes in melanin content were identified, and 4 proteins, namely, Wnt8a, Slc27a, Mucin-2 (Muc2), and gamma-glutamyl transferase (Ggt5), were selected for validation via Western blotting. Metabolomic analysis revealed strong correlations between L-tyrosine, 4-N-methyltransferase, indole-3-acetate salt, L-phenylalanine, L-threonine, L-glutamic acid, indole, and skin color formation in T. fasciatus. Notably, joint proteomic and metabolomic analyses revealed significant enrichment of ABC transporters across all three skin colors of T. fasciatus. Additionally, we identified 25 differentially expressed metabolites, suggesting potential feed additives such as adrenosterone and 1-(3-ethyl-2,4-dihydroxy-6-methoxyphenyl) butan-1-one for adjusting skin color. This research enhances our understanding of fish skin color formation mechanisms and provides valuable insights for cultivating fish with specific skin colors.
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