The Function of the Lysophospholipase 1 (LYPLA1) Gene in Fetal Sheep Myoblast Proliferation and Myogenic Differentiation

Evidence from genome-wide association studies (GWAS) links genetic variants near the <i>LYPLA1</i> locus to variations in sheep muscle development. To further investigate whether <i>LYPLA1</i> expression plays a role in fetal sheep myoblast development, we performed gain-of-function/loss-of-function analyses targeting <i>LYPLA1</i> using siRNA and recombinant vectors. CCK-8 assays, EdU staining, and flow cytometry were used to investigate the effects of both <i>LYPLA1</i> knockdown and overexpression on the proliferation of fetal sheep myoblasts. Furthermore, this study investigated the effect of <i>LYPLA1</i> on myogenic differentiation by means of indirect immunofluorescence staining with MyHC and quantitative analysis of myogenic regulatory factor (MRF) expression. The results showed that downregulation of <i>LYPLA1</i> significantly reduced cell viability and proliferation rate, arrested myoblasts in the G2 phase, and inhibited proliferation. These effects were reversed when <i>LYPLA1</i> was overexpressed. Furthermore, <i>LYPLA1</i> knockdown significantly impaired myotube formation during terminal differentiation (days 5-7). This defect was correlated with a marked reduction in the expression of myogenic regulatory factors (MRFs). This effect was reversed when <i>LYPLA1</i> was highly expressed. In summary, this study demonstrates that <i>LYPLA1</i> promotes the proliferation and differentiation of fetal sheep myoblasts, establishing its potential as a target for the molecular breeding of sheep.