LDL和PCS对山羊精子冷冻效果及脂类含量影响的研究
摘要
由于山羊冷冻精液的受精率低于自然交配,还没有在生产中推广,山羊冷冻精液受精率低的原因之一可能是在冷冻过程中精子的胆固醇流失,过早获能造成的。因此,本文以绒山羊为实验动物,研究在稀释液中添加低密度脂蛋白(LDL)和胆固醇硫酸脂(PCS)对山羊精子胆固醇和磷脂含量以及冷冻保存效果的研究。首先用LDL代替卵黄去探讨对精子冷冻保护的作用,然后进一步添加PCS,研究在冷冻过程中LDL和PCS对精子胆固醇的保护作用及对精子的活力、膜完整率和顶体完整率的影响。
1.在稀释液中以LDL替代卵黄后,LDL可以维持精液冷冻过程中山羊精子的胆固醇含量和胆固醇与磷脂的比例,能提高冷冻—解冻后山羊精子的活率、质膜完整率和顶体完整率。因此,LDL可以替代卵黄作为山羊精液冷冻的保护剂。LDL的添加量以9gLDL/100ml对精子的保护作用最好。
2.在LDL稀释液中添加PCS,可提高冷冻后山羊精子胆固醇与磷脂的比值,可进一步提高山羊精液冷冻后精子的活率和质膜完整率,而对精子顶体无不良影响。因此,在山羊LDL精液稀释液中添加PCS是可行的。
3.在EY稀释液中添加PCS可提高山羊精子冷冻后的磷脂含量,避免山羊精子在冷冻过程中胆固醇的流失,维持冷冻过程中精子胆固醇与磷脂的比例,提高精子活率、精子质膜完整率和精子顶体完整率(I型顶体),可以作为山羊精液冷冻稀释液的成分。添加量以0.10gPCS/100ml为宜。
For the lower pregnant rate of AI with goat frozen semen than that of natural mating, the goat frozen semen were still not used commercially. One of the reasons of lower pregnant rate might be the depletion of cholesterol from goat sperm during freezing leading to premature capacitation. Therefore, the paper studied on the effect of low density lipoprotein(LDL) and potassium cholesterol sulfate(PCS) to the extender on the cholestero, phospholipids contents of goat spermatozoa and their cryopreservation during Freezing Process. Firstly,LDL was used to instead of egg yolk in diluents to study on the protection of sperm from freeze. Additional, PCS was supplemented to LDL diluent and EY diluent to study on the ability of LDL+PCS and EY+PCS to maintain the cholesterol and phospholipids contents, motility, membrane and acrosome of sperm.
1.When EY was replaced with LDL in goat semen frozen diluent, LDL could maintain the cholesterol content and the ratio of cholesterol and phospholipids. LDL could also increase the motility, the rates of no-effected embrane and acrosome of frozen goat sperm.So, LDL could be used to replace the EY in diluent, and the suitable dosage was 9g LDL in 100ml diluent.
2.When PCS was added to the LDL diluent, PCS could increase the ratio of cholesterol and pgospholipid, motility and the rate of no-effected membrane of frozen goat sperm. PCS could be used as a composition in goat semen frozen diluent.
3.In EY diluent, PCS could increase the ratio of cholesterol and phospholipid, motility and the rate of no-effected membrane of goat sperm. So, PCS could be used as a composition in gaot semen frozen diluent and suitable dosage of PCS was 0.10g/100ml diluent.
1.在稀释液中以LDL替代卵黄后,LDL可以维持精液冷冻过程中山羊精子的胆固醇含量和胆固醇与磷脂的比例,能提高冷冻—解冻后山羊精子的活率、质膜完整率和顶体完整率。因此,LDL可以替代卵黄作为山羊精液冷冻的保护剂。LDL的添加量以9gLDL/100ml对精子的保护作用最好。
2.在LDL稀释液中添加PCS,可提高冷冻后山羊精子胆固醇与磷脂的比值,可进一步提高山羊精液冷冻后精子的活率和质膜完整率,而对精子顶体无不良影响。因此,在山羊LDL精液稀释液中添加PCS是可行的。
3.在EY稀释液中添加PCS可提高山羊精子冷冻后的磷脂含量,避免山羊精子在冷冻过程中胆固醇的流失,维持冷冻过程中精子胆固醇与磷脂的比例,提高精子活率、精子质膜完整率和精子顶体完整率(I型顶体),可以作为山羊精液冷冻稀释液的成分。添加量以0.10gPCS/100ml为宜。
For the lower pregnant rate of AI with goat frozen semen than that of natural mating, the goat frozen semen were still not used commercially. One of the reasons of lower pregnant rate might be the depletion of cholesterol from goat sperm during freezing leading to premature capacitation. Therefore, the paper studied on the effect of low density lipoprotein(LDL) and potassium cholesterol sulfate(PCS) to the extender on the cholestero, phospholipids contents of goat spermatozoa and their cryopreservation during Freezing Process. Firstly,LDL was used to instead of egg yolk in diluents to study on the protection of sperm from freeze. Additional, PCS was supplemented to LDL diluent and EY diluent to study on the ability of LDL+PCS and EY+PCS to maintain the cholesterol and phospholipids contents, motility, membrane and acrosome of sperm.
1.When EY was replaced with LDL in goat semen frozen diluent, LDL could maintain the cholesterol content and the ratio of cholesterol and phospholipids. LDL could also increase the motility, the rates of no-effected embrane and acrosome of frozen goat sperm.So, LDL could be used to replace the EY in diluent, and the suitable dosage was 9g LDL in 100ml diluent.
2.When PCS was added to the LDL diluent, PCS could increase the ratio of cholesterol and pgospholipid, motility and the rate of no-effected membrane of frozen goat sperm. PCS could be used as a composition in goat semen frozen diluent.
3.In EY diluent, PCS could increase the ratio of cholesterol and phospholipid, motility and the rate of no-effected membrane of goat sperm. So, PCS could be used as a composition in gaot semen frozen diluent and suitable dosage of PCS was 0.10g/100ml diluent.
引文
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2 胡建宏.猪精液冷冻保存研究.博士研究生学位论文.2006 年
3 Polge C.Artificial insemination in pigs[J].Vet Pec,1956,68:62-76
4 Pursel VG,Johnson LA.Freezing of boar spermatozoa: Fertilizing capacity with concentrated semen anda new thawing procedure[J]. J Animal Sci,1975,40:99-102
5 陶勇,经荣斌.家畜精液冷冻保存的研究综述. 国外畜牧学-猪与禽,1999,6:39-42
6 Rodriguez-Martinez H, Eriksson B,Lundeheim. Freezing boar semen in flat plastic bags[J].Membraneintegrity and fertility[J].Reproduction Fertility Development,1996,31:161-168
7 Dass den JHG, Jong de G,Lansbergen LM,et al. Capacitation as a regulatory event that primesspermatozoa for the acrosome reaction and fertilization [J]. Molecular Human Reproduction,1997,3:175–194
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