Supplementation of L-ascorbic acid improves the in vitro development of buffalo(Bubalus bubalis)embryos and alters the expression of apoptosis-related genes

摘要

Objective:To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic,anti-apoptotic,and embryonic development-related genes.Methods:In experiment 1,we evaluated the effect of the addition of 0(control),50,and 100μM L-ascorbic acid to the in vitro maturation medium on the developmental competence in terms of blastocyst rate and relative mRNA abundance of some pro-apoptotic(BAX,BID),anti-apoptotic(BCL-XL,MCL1),and embryonic development(GDF9,BMP15)related genes.Based on the results,we chose 50μM as the suitable dose of L-ascorbic acid for the subsequent experiments.We further evaluated the blastocyst rates following the addition of 50μM L-ascorbic acid to the in vitro culture medium(experiment 2),and in vitro maturation and in vitro culture media(experiment 3).In all three experiments,the maturation and culture media devoid of L-ascorbic acid served as the control group.Results:The blastocyst rate after adding 50μM L-ascorbic acid to the in vitro maturation medium was significantly higher than the control group(P<0.05),whereas 100μM L-ascorbic acid exhibited a negative effect on the blastocyst rate.The blastocyst rates for embryos cultured in 50μM L-ascorbic acid in the in vitro culture medium alone and both in vitro maturation and in vitro culture media were significantly higher than their corresponding control groups(P<0.05).The relative mRNA abundance of BAX significantly decreased in blastocysts produced after the addition of 50μM L-ascorbic acid as compared with the control group(P<0.05),whereas,for MCL1,it significantly decreased in blastocysts produced after the addition of 100μM L-ascorbic acid(P<0.05).Conclusions:The supplementation of 50μM L-ascorbic acid to in vitro maturation and in vitro culture media supports in vitro embryonic development in buffaloes by improving developmental competence and altering the expression of apoptosis-related genes.