Lipid-Enriched Diet Advances Early Oocyte Development in the Endangered Leptobotia elongata: Metabolomics-Based...

Leptobotia elongata, an endangered freshwater fish endemic to the upper Yangtze River, increasingly depends on artificial reproduction for conservation. However, a captive juvenile population fed a conventional diet commonly exhibits ovarian development arrested at stages I-II, representing a critical bottleneck for captive reproduction. To address this, juvenile L. elongata were reared for 6 months on either a basal diet (control group, CG) or a lipid-enriched diet in which soybean oil was replaced by fish oil and soybean lecithin, with vitamin E added (treatment group, TG). Histological examination revealed that TG ovaries advanced from stage I-II to stage III, with markedly larger oocytes and centralized nucleoli. LC-MS/MS-based untargeted metabolomics profiling identified 1773 metabolites, of which 566 differed significantly between groups (374 up- and 192 down-regulated in TG). KEGG enrichment revealed 15 significantly perturbed pathways, with nucleotide metabolism showing the highest enrichment (Rich factor = 0.22), accompanied by changes in fatty acid biosynthesis, biosynthesis of unsaturated fatty acids, glycerophospholipid metabolism, aminoacyl-tRNA biosynthesis, pantothenate and CoA biosynthesis, and TCA-related modules. Coordinated up-regulation of long-chain polyunsaturated fatty acids (DHA, EPA, ARA, α-linolenic acid), DHA-phosphatidylethanolamine, phosphatidylcholine and CDP-choline, together with depletion of their precursors (sn-glycerol-3-phosphate, LPI(20:4), pyrophosphate), was consistent with enhanced phospholipid remodeling and Lands-cycle activity. A multi-tier regulatory network was constructed, anchored on three pathway hubs (nucleotide metabolism, pyrimidine metabolism and fatty acid elongation) and five functional modules. This network indicated that dietary lipid supplementation was associated with coordinated changes across four interconnected metabolic axes: lipid supply and membrane remodeling, nucleotide synthesis and turnover, amino acid-protein translation, and energy-coenzyme metabolism. Collectively, these findings propose a metabolic framework and identify candidate biomarkers (e.g., DHA-PE, PC) for optimizing feed formulations of L. elongata.