鲤lpl-a序列特征及其过表达对脂肪变化的影响OA

To investigate the effects of overexpressing lipoprotein lipase a gene(lpl-a)in common carp(Cyprinus carpio)on fat deposition patterns and lipid metabolism-related genes expression,we analyzed the sequence and struc-tural characteristics of the carp lpl-a and examined its tissue distribution using quantitative Real-time(qPCR)in five internal tissues:abdominal fat,liver,kidney,muscle,and intestine.The lpl-a overexpression plasmid vector was constructed by homologous recombination and introduced into zebrafish(Danio rerio)and carp liver cells.The impacts of lpl-a overexpression on total cholesterol(T-CHO),triglyceride(TG)levels,and expressions of lipid metabolism marker genes(ppar γ,fabp 3,pnpla 2,etc.)were assessed in zebrafish(individual level)and carp hepatocytes(cell level).Results showed that the carp lpl-a was predicted to encode a 507-amino acid protein with 7 open reading frames(ORFs),and it demonstrated high evolutionary conservation in osteichthyes.Tissue-specific expression analysis revealed significantly higher lpl-a transcript levels in abdominal fat,liver,and kidney compared to muscle and intestine(P<0.05).In zebrafish,microinjection of the lpl-a overexpression vector resulted in an approximately 7-fold increase in muscle T-CHO and a 1-fold increase in both T-CHO and TG in liver,accompanied by upregulation of lpl-a,ppar γ,and fabp 3(P<0.05).In carp hepatocytes,transfection with the lpl-a construct for 72h significantly increased lpl-a and lipolytic gene pnpla 2 expressions(P<0.05).Altogether,these findings demonstrate that overexpression of lpl-a induced distinct lipid metabolism regulations at individual(enhanced muscle fat storage and hepatic lipid accumulation)and cellular(activated lipolysis and inhibited lipogenesis)levels,providing theoretical insights for targeting lpl-a to modulate lipid deposition in economically important aquatic animals.