Jordi Roca, Maria A. Gil, Marta Hernandez, Inma Parrilla, Juan M. Vazquez, Emilio A. Martinez
{"title":"猪精子在添加丁基羟基甲苯的膨化剂中冻融后的存活率和育性","authors":"Jordi Roca, Maria A. Gil, Marta Hernandez, Inma Parrilla, Juan M. Vazquez, Emilio A. Martinez","doi":"10.1002/j.1939-4640.2004.tb02806.x","DOIUrl":null,"url":null,"abstract":"<p><b>ABSTRACT: </b> This study evaluated the protective effect of butylated hydroxytoluene (BHT), a lipid-soluble antioxidant, against cryopreservation injuries to boar spermatozoa. In experiment 1, the lowest BHT concentrations able to reduce lipid peroxidation in boar spermatozoa were determined. Nine BHT concentrations (ranging from 0.025 to 3.2 mM) were evaluated, and the lowest (P < .05) production of malondialdehyde (MDA), as an indicator of lipid peroxidation, was obtained when BHT ranged from 0.2 to 1.6 mM. In experiment 2, sperm survivability was evaluated when BHT was added to a postthaw freezing extender by measuring the degree of sperm lipid peroxidation (using MDA production) and by measuring parameter such as motility, plasma membrane and acrosome integrity, and cell apoptosis. The ability of thawed spermatozoa to fertilize in vitro—matured oocytes and of embryos to develop to the blastocyst stage in vitro was also assessed. Pooled sperm-rich fractions collected from 3 mature Pietrain boars were frozen in 0.5-mL straws after dilution with lactose-egg yolk-glycerol-Orvus ES Paste extender supplemented with 0, 0.2, 0.4, 0.8, and 1.6 mM BHT. Postthaw sperm survival, evaluated 30 and 150 minutes after thawing, was higher in BHT-treated spermatozoa, being significant (P < .05) when the freezing extender was supplemented with 0.2, 0.4, and 0.8 mM BHT. The addition of BHT to the freezing extender resulted in a significant (P < .05) decrease in the MDA concentration in thawed spermatozoa, irrespective of the level of BHT used. BHT had no effect on oocyte cleavage rates, but the development to blastocyst was improved for embryos derived from spermatozoa frozen in extender supplemented with 0.4 mM BHT (16% vs 29% of blastocysts per total oocytes; P < .05). In conclusion, under the conditions tested in the present study, the addition of BHT to the freezing extender improved the overall efficiency of thawed boar spermatozoa.</p>","PeriodicalId":15029,"journal":{"name":"Journal of andrology","volume":"25 3","pages":"397-405"},"PeriodicalIF":0.0000,"publicationDate":"2013-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/j.1939-4640.2004.tb02806.x","citationCount":"178","resultStr":"{\"title\":\"Survival and Fertility of Boar Spermatozoa After Freeze-Thawing in Extender Supplemented With Butylated Hydroxytoluene\",\"authors\":\"Jordi Roca, Maria A. Gil, Marta Hernandez, Inma Parrilla, Juan M. Vazquez, Emilio A. 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The ability of thawed spermatozoa to fertilize in vitro—matured oocytes and of embryos to develop to the blastocyst stage in vitro was also assessed. Pooled sperm-rich fractions collected from 3 mature Pietrain boars were frozen in 0.5-mL straws after dilution with lactose-egg yolk-glycerol-Orvus ES Paste extender supplemented with 0, 0.2, 0.4, 0.8, and 1.6 mM BHT. Postthaw sperm survival, evaluated 30 and 150 minutes after thawing, was higher in BHT-treated spermatozoa, being significant (P < .05) when the freezing extender was supplemented with 0.2, 0.4, and 0.8 mM BHT. The addition of BHT to the freezing extender resulted in a significant (P < .05) decrease in the MDA concentration in thawed spermatozoa, irrespective of the level of BHT used. BHT had no effect on oocyte cleavage rates, but the development to blastocyst was improved for embryos derived from spermatozoa frozen in extender supplemented with 0.4 mM BHT (16% vs 29% of blastocysts per total oocytes; P < .05). 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引用次数: 178
摘要
摘要:本研究探讨了脂溶性抗氧化剂丁基羟基甲苯(BHT)对猪精子低温保存损伤的保护作用。在实验1中，确定了降低猪精子脂质过氧化的最低BHT浓度。9种BHT浓度(范围从0.025到3.2 mM)被评估，最低(P <当BHT在0.2 ~ 1.6 mM范围内时，获得了作为脂质过氧化指标的丙二醛(MDA)的产生。在实验2中，通过测量精子脂质过氧化程度(利用MDA的产生)和测量运动性、质膜和顶体完整性以及细胞凋亡等参数，在解冻后的冷冻扩展剂中添加BHT来评估精子的存活能力。解冻的精子在体外成熟的卵母细胞中受精的能力和胚胎在体外发育到囊胚期的能力也被评估。从3头成熟的Pietrain公猪身上收集富精子的混合部分，用添加0、0.2、0.4、0.8和1.6 mM BHT的乳糖-蛋黄-甘油- orvus ES糊剂稀释后，冷冻在0.5 ml吸管中。解冻后30分钟和150分钟的精子存活率，bht处理的精子更高，具有显著性(P <分别添加0.2、0.4和0.8 mM BHT时，冷冻扩展剂的浓度为0.05)。在冻结剂中加入BHT可显著降低(P <.05)解冻精子中MDA浓度的降低，与BHT的使用水平无关。BHT对卵母细胞的分裂率没有影响，但在添加0.4 mM BHT的培养基中冷冻的精子胚胎向囊胚的发育有所改善(囊胚占卵母细胞总数的16% vs 29%);P & lt;. 05)。综上所述，在本试验条件下，冷冻膨化剂中添加BHT可提高猪精子解冻的整体效率。
Survival and Fertility of Boar Spermatozoa After Freeze-Thawing in Extender Supplemented With Butylated Hydroxytoluene
ABSTRACT: This study evaluated the protective effect of butylated hydroxytoluene (BHT), a lipid-soluble antioxidant, against cryopreservation injuries to boar spermatozoa. In experiment 1, the lowest BHT concentrations able to reduce lipid peroxidation in boar spermatozoa were determined. Nine BHT concentrations (ranging from 0.025 to 3.2 mM) were evaluated, and the lowest (P < .05) production of malondialdehyde (MDA), as an indicator of lipid peroxidation, was obtained when BHT ranged from 0.2 to 1.6 mM. In experiment 2, sperm survivability was evaluated when BHT was added to a postthaw freezing extender by measuring the degree of sperm lipid peroxidation (using MDA production) and by measuring parameter such as motility, plasma membrane and acrosome integrity, and cell apoptosis. The ability of thawed spermatozoa to fertilize in vitro—matured oocytes and of embryos to develop to the blastocyst stage in vitro was also assessed. Pooled sperm-rich fractions collected from 3 mature Pietrain boars were frozen in 0.5-mL straws after dilution with lactose-egg yolk-glycerol-Orvus ES Paste extender supplemented with 0, 0.2, 0.4, 0.8, and 1.6 mM BHT. Postthaw sperm survival, evaluated 30 and 150 minutes after thawing, was higher in BHT-treated spermatozoa, being significant (P < .05) when the freezing extender was supplemented with 0.2, 0.4, and 0.8 mM BHT. The addition of BHT to the freezing extender resulted in a significant (P < .05) decrease in the MDA concentration in thawed spermatozoa, irrespective of the level of BHT used. BHT had no effect on oocyte cleavage rates, but the development to blastocyst was improved for embryos derived from spermatozoa frozen in extender supplemented with 0.4 mM BHT (16% vs 29% of blastocysts per total oocytes; P < .05). In conclusion, under the conditions tested in the present study, the addition of BHT to the freezing extender improved the overall efficiency of thawed boar spermatozoa.
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