精子选择方法对牛精子质量及体外受精效果的影响  

Effects of sperm selection methods on bovine sperm quality and in vitro fertilization outcomes

作　　者：张楠 郭桃 韩僖彤 张建军 张家新 ZHANG Nan;GUO Tao;HAN Xitong;ZHANG Jianjun;ZHANG Jiaxin(Inner Mongolia Key Laboratory of Sheep&Goat Genetics,Breeding and Reproduction,Hohhot 010018,China;College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古自治区羊遗传育种与繁殖重点实验室,内蒙古呼和浩特010018 [2]内蒙古农业大学动物科学学院,内蒙古呼和浩特010018

出　　处：《黑龙江动物繁殖》2025年第1期24-30,36,共8页Heilongjiang journal of animal reproduction

基　　金：内蒙古自治区揭榜挂帅项目(2022JBGS0025)。

摘　　要：为了探究不同精子选择方法对精子质量及体外受精(IVF)后胚胎发育的影响,试验以牛冷冻精液为研究对象,使用TALP液直接离心法(对照组)、TALP-Percoll-密度梯度离心法(1组)、TALP-PureSperm-密度梯度离心法(2组)处理牛冷冻精液,然后分别评价处理后精液的精子活力、运动参数[前向运动精子比例、平均路径速度(VAP)、直线运动速度(VSL)、曲线运动速度(VCL)、直线度(STR)、头部横向位移幅度(ALH)]、质膜完整率、顶体膜完整率、线粒体膜电位水平、活性氧(ROS)浓度、DNA完整性、获能精子率、顶体反应精子率、酪氨酸磷酸化水平,并比较精子IVF后胚胎发育情况(卵裂率、囊胚率)。结果表明:1组和2组的精子活力、前向运动精子比例、VAP、VSL、VCL、STR、ALH、精子质膜完整率、顶体反应精子率均显著高于对照组(P<0.05),精子DNA完整率高于对照组(P>0.05),精子ROS含量显著低于对照组(P<0.05),其中除ROS含量外,1组的其他指标均高于2组,其中精子活力差异显著(P<0.05),其他指标两组间差异不显著(P>0.05);1组和对照组的精子顶体完整率、精子酪氨酸磷酸化水平显著高于2组(P<0.05),1组和对照组间差异不显著(P>0.05);1组精子线粒体膜电位、获能精子率显著高于对照组和2组(P<0.05);1组卵裂率和囊胚率显著高于对照组(P<0.05),也高于2组,但差异不显著(P>0.05)。说明牛冷冻精液经TALP-Percoll-密度梯度离心法选择出的精子质量较优,且体外受精后胚胎发育效果更好,可作为牛IVF精子选择的首选方法。To explore the effects of different sperm selection methods on sperm quality and embryo development after in vitro fertilization(IVF),bovine frozen semen was treated with the TALP liquid direct centrifugation method(control group),TALP Percoll density gradient centrifugation(group 1)and TALP PureSperm density gradient centrifugation(group 2)to evaluate the sperm motility,motility parameters(forward motile sperm ratio,average path velocity(VAP),linear velocity(VSL),curvilinear velocity(VCL),straightness(STR),head lateral displacement amplitude(ALH),plasma membrane integrity,acrosomal membrane integrity,mitochondrial membrane potential level,reactive oxygen species(ROS),concentration,DNA integrity,capacitated sperm rate,acrosome reaction sperm rate,tyrosine phosphorylation level and compare the embryonic development after IVF(cleavage rate,blastocyst rate)in different treatment groups.The results showed that the sperm motility,the proportion of forward motile sperm,the percentage of forward motile sperm and the percentage of forward motile sperm,VAP,VSL,VCL,STR,ALH,the sperm plasma membrane integrity rate and acrosome reaction sperm rate were significantly higher than those in the control group(P<0.05).The sperm DNA integrity rate was higher than that in the control group(P>0.05),and the sperm ROS content was significantly lower than that in the control group(P<0.05).Except for ROS content,other indicators in group 1 were higher than those in group 2,in which the difference in sperm motility was significant(P<0.05),while no significant difference in other indicators between the two groups(P>0.05).The sperm acrosomal membrane integrity rate and sperm tyrosine phosphorylation level in group 1 and in control group were significantly higher than those in group 2(P<0.05),while no significant difference between group 1 and control group(P>0.05).Sperm mitochondrial membrane potential and capacitated sperm rate in group 1 were significantly higher than those in control group and in group 2(P<0.05);The cleavage rate and bla

关 键 词：牛 体外受精 精子选择 精子获能 PERCOLL PureSperm 胚胎发育 

分 类 号：S823.34[农业科学—畜牧学]