Multi-omics analysis and longitudinal study of reprogramming by dietary creatine to endogenous metabolism in largemouth bass (Micropterus salmoides)

Creatine is a feed additive with physiological pleiotropic properties and also an energy homeostasis protector in vertebrates and is successfully used in terrestrial livestock and aquaculture. Here, two feeding trials were performed to investigate dietary creatine on endogenous creatine metabolism and physiological reprogramming in largemouth bass. The results showed that the endogenous creatine metabolism genes AGAT, GAMT, and SLC6A8 of largemouth bass are highly conserved with the amino acid sequences of other teleosts and are clustered separately from mammals. Among the 16 major tissues in largemouth bass, both creatine synthesis genes (agat, gamt) and transporter gene slc6a8 are most highly expressed in muscle. Muscle has a high threshold but sensitive creatine negative feedback to regulate endogenous creatine metabolism. Dietary creatine intake significantly inhibits endogenous creatine synthesis and transport in muscle in a dose-dependent manner, and this inhibitory effect recovers with a decrease in dietary creatine content. In addition, physiological creatine saturation required prolonged exogenous creatine intake, and it would be shortened by high doses of creatine, which provides guidance for maximizing economic benefits in aquaculture. Metabolome and transcriptome showed that dietary creatine significantly affected the metabolism of the creatine precursor substance-arginine. Exogenous creatine intake spared arginine that would otherwise be used for creatine synthesis, increased arginine levels, and caused reprogramming of arginine metabolism. Overall, these results demonstrate that the addition of creatine to largemouth bass diets is safe and recoverable, and the benefits of creatine intake in largemouth bass are not limited to enhancing the function of creatine itself but also include a reduction in the metabolic burden of essential amino acids to better growth performance.