Integration analysis of metabolome and transcriptome reveals the effect of exogenous supplementation with mixtures of vitamins ADE, zinc, and selenium on follicular growth and granulosa cells molecular metabolism in donkeys (Equus asinus)

Vitamins and microelements play essential roles in mammalian ovarian physiology, including follicle development, ovulation, and synthesis and secretion of hormones and growth factors. However, it is nevertheless elusive to what extent exogenous supplementation with mixtures of vitamins ADE, zinc (Zn), and selenium (Se) affects follicular growth and granulosa cells (GCs) molecular function. We herein investigated their effect on follicular growth and GCs physiological function. We showed that follicular growth and ovulation time was accelerated and shortened with the increases of vitamins ADE, Zn, and Se doses by continually monitoring and recording (one estrus cycle of about 21 days) with an ultrasound scanner. Integrated omics analysis showed that there was a sophisticated network relationship, correlation expression, and enrichment pathways of the genes and metabolites highly related to organic acids and their derivatives and lipid-like molecules. Quantitative real-time PCR (qPCR) results showed that vitamin D receptor ( VDR ), transient receptor potential cation channel subfamily m member 6 ( TRPM6 ), transient receptor potential cation channel subfamily v member 6 ( TRPV6 ), solute carrier family 5 member 1 ( SLC5A1 ), arachidonate 5-lipoxygenase ( ALOX5 ), steroidogenic acute regulatory protein ( STAR ), prostaglandin-endoperoxide synthase 2 ( PTGS 2), and insulin like growth factor 1 ( IGF-1 ) had a strong correlation between the transcriptome data. Combined multi-omics analysis revealed that the protein digestion and absorption, ABC transporters, biosynthesis of amino acids, aminoacyl-tRNA biosynthesis, mineral absorption, alanine, aspartate and glutamate metabolism, glycine, serine and threonine metabolism, arginine biosynthesis, and ovarian steroidogenesis were significantly enriched. We focused on the gene-metabolite interactions in ovarian steroidogenesis, founding that insulin receptor ( INSR ), phospholipase a2 group IVA ( PLA2G4A ), adenylate cyclase 6 ( ADCY6 ), cytochrome p450 family 1 subfamily b member 1 ( CYP1B1 ), protein kinase camp-activated catalytic subunit beta ( PRKACB ), cytochrome p450 family 17 subfamily a member 1 ( CYP17A1 ), and phospholipase a2 group IVF ( PLA2G4F ) were negatively correlated with β-estradiol (E 2 ), progesterone (P 4 ), and testosterone (T) ( P < 0.05). while ALOX5 was a positive correlation with E2, P 4 , and T ( P < 0.05); cytochrome p450 family 19 subfamily a member 1 ( CYP19A1 ) was a negative correlat