show Abstracthide AbstractRNA from buffalo immature Leydig cells (ILCs) and adult Leydig cells (ALCs) were isolated and sequenced using BGI DNBseq. We found that development of Leydig cells from ILCs to ALCs was associated with decreased expression of genes related to cell cycle, cell junction, adhesion and increased expression of genes related to steroidogenesis and lipid transport. Based on functional enrichment analysis and network analysis, cAMP pathway may play a key role in the testosterone function of the buffalo was characterized during ILC-to-ALC transition. In brief, our discoveries will enhance our understanding of developmental mechanism studies related to buffalo Leydig cells testosterone function.