精浆对猪精子冷冻效果影响的研究

›› 2012, Vol. 39 ›› Issue (11): 161-164.

精浆对猪精子冷冻效果影响的研究

刘彪, 逯心玉, 孔庆然, 苏兴盼, 何文腾, 胡魁, 刘忠华

东北农业大学胚胎工程实验室,黑龙江哈尔滨 150030

修回日期:2012-05-15 出版日期:2012-11-20 发布日期:2012-11-22
通讯作者: 刘忠华,教授,博士生导师,研究方向:克隆与转基因猪,早期胚胎发育机理。Tel:0451-55191729;E-mail:liu086@yahoo.com E-mail:liu086@yahoo.com
作者简介:刘彪(1985-),男,河南人,硕士生,主要从事猪冷冻精液的研究。
基金资助:
国家重大基础科学研究项目(973)资助(2009CB941002);黑龙江省杰出青年基金(JC200905)。

Study on the Effect of Seminal Plasma in Porcine Sperms Cryopreservation

LIU Biao, LU Xin-yu, KONG Qing-ran, SU Xing-pan, HE Wen-teng, HU Kui, LIU Zhong-hua

Laboratory of Embryo Engineering, Northeast Agricultural University,Harbin 150030,China

Revised:2012-05-15 Online:2012-11-20 Published:2012-11-22

摘要/Abstract

摘要： 为优化猪精子冷冻技术,提高解冻后精子的活力和受精能力,本试验分别以含精浆浓度为10%、20%和30%的冷冻保护剂处理精子,以冷冻前、解冻后精子活力和质膜完整性,解冻后精子进行体外受精(IVF)的卵裂率和囊胚率等作为检测指标,同时以含有卵黄的Tris-柠檬酸－葡萄糖(Tris-citric acid-glucose,TCG)冷冻基础液作为对照研究精浆对猪冷冻精子的保护作用。结果显示,在含有10%精浆浓度的稀释液中,冷冻前质膜完整性,解冻后精子活率、质膜完整性、IVF囊胚率相对于对照组均显著提高(P<0.05);当含有10%精浆的冷冻精液解冻后用于人工授精时,与配母猪妊娠率、窝产仔数、窝产活仔数等仍显著低于鲜精授精组(P<0.05)。上述结果表明,含10%精浆的冷冻保护剂能提高精子的冷冻后活力和IVF胚胎发育率,但用于人工授精配种与鲜精相比还有一定差距。

关键词: 猪精子; 精浆; 冷冻保存; 体外受精; 人工授精

Abstract: In order to improve the frozen porcine sperms motility and fertilizing ability after thawed, sperms were extended in basic cryoprotection extender with 0, 10%, 20%, 30% seminal plasma respectively before freezing. Sperm motility, membrane integrity, fertilization rate and blastocyst rate of IVF embryos and artificial insemination results were chosen as criteria for assessing the sperm quality. The results showed that the motility, membrane integrity of sperms extended in cryoprotection extender containing 10% seminal plasma were improved significantly (P<0.05) after thawed compared with the control group. The blastocyst rate of IVF embryos derived from these sperms was also increased significantly. Artificial insemination with the thawed sperms from the group extended in cryoprotection extender containing 10% seminal plasma resulted in a significantly lower pregnancy rates, litter size and number of live littermate compared with fresh sperm insemination(P<0.05). These results indicated that cryoprotectant with 10% seminal plasma could increase the motility of thawed porcine sperm after cryopreservation and the blastocyst rate after IVF, but for artificial insemination it was still need to be improved.

Key words: porcine sperm; seminal plasma; cryopreservation; IVF; artificial insemination

中图分类号:

S814

刘彪, 逯心玉, 孔庆然, 苏兴盼, 何文腾, 胡魁, 刘忠华. 精浆对猪精子冷冻效果影响的研究[J]. , 2012, 39(11): 161-164.

LIU Biao, LU Xin-yu, KONG Qing-ran, SU Xing-pan, HE Wen-teng, HU Kui, LIU Zhong-hua. Study on the Effect of Seminal Plasma in Porcine Sperms Cryopreservation[J]. , 2012, 39(11): 161-164.

参考文献

1 杨军祥. 除去精浆对波尔山羊冷冻精液品质的影响[J]. 中国草食动物,2009,29(2):22~24.
2 滕尚辉,宋成义. 猪PSP-Ⅰ/PSP-Ⅱ的生物学特性研究[J]. 上海畜牧兽医通讯,2007,5:54~55.
3 Caballero I,Vázquez J M,Centurión F,et al. Comparative effects of autologous and homologous seminal plasma on the viability of largely extended boar spermatozoa[J]. Reprod Domest Anim,2004,39:370~375.
4 Calvete J J,Mann K,Schafer W,et al. Boar spermadhesin PSP-Ⅱ:Location of posttranslational modifications,heterodimer formation with PSP-Ⅰ glycoforms and effect ofdimerization on the ligand-binding capabilities of the subunits[J]. FEBS Lett,1995,365:179~182.
5 Cosentino M J,Emilson L B V,Cockett A T K. Prostaglandins in semen and their relationship to male fertility:A study of 145 men[J]. Fertil S teril,1984,41:88.
6 Flesh F M,Gadella B M. Dynamics of the mammalian sperm plasma membrane in the process of fertilization[J]. Biochim Biophys Acta,2000,1469(3):197~235.
7 Fraser L, et al. Dialysis of boar semen prior to freezing-thawing:Its effects on post-thaw sperm characteristics[J]. Theriogenology,2007,56:994~1003.
8 Garciaa J C,Domingueza J C,Penac F J,et al. Thawing boar semen in the presence of seminal plasma:Effects on sperm quality and fertility[J]. Animal Reproduction Science,2010,119:160~165.
9 Larson J L,Miller D J.Simple histo-chemical stain for acrosomes on sperm from several species[J].Mol Reprod Dev,1999,52: 445~449.
10 Lindholmer C H. The importance of seminal plasma for human sperm motility[J]. Bio Reprod,1974,10:533.
11 Martine P F,et al. Seminal plasma improves cryopreservation of iberian red deer epididymal sperm[J]. Theriogenology,2006,66:1847~1856.
12 Polge C. Artificial insemination in pigs[J] . Vet Rec,1956, 68: 62~76.
13 Strzezek J,Wysocki P,Kordan W,et al. Proteomics of boar seminal plasma-current studies and possibility of their application in biotechnology of animal reproduction[J]. Reprod Biol,2005,5:279~290.

[1]	张笑梦, 焦安惠, 王玉琪, 方南洙, 金庆国. 添加罗格列酮对小鼠卵母细胞氧化还原稳态的影响[J]. 中国畜牧兽医, 2023, 50(5): 1818-1827.
[2]	张雨阳, 曲星霖, 陈璇, 吕艳秋, 曹立朋, 张君正, 金一. 泛素激活酶抑制剂对猪精子获能状态、膜重构及体外受精的影响[J]. 中国畜牧兽医, 2023, 50(1): 210-219.
[3]	梁家充, 吕春荣, 洪琼花, 吴国权, 权国波. 云南半细毛羊精子冷冻前后转录组表达差异分析[J]. 中国畜牧兽医, 2022, 49(3): 973-981.
[4]	王轶男, 孔令敏, 徐海峰, 张正文, 李春宇, 尹伊, 金一, 吕艳秋. 解冻稀释液中添加精浆对冻融猪精子品质的影响[J]. 中国畜牧兽医, 2022, 49(2): 615-623.
[5]	薄欣宇, 陶晓钊, 任战军, 董响贵, 王淑辉. β-神经生长因子在家兔诱导排卵中的研究进展[J]. 中国畜牧兽医, 2022, 49(11): 4288-4295.
[6]	李梦杰, 赵伟刚, 郭肖兰, 顾士钢, 赵全民, 徐超. 冷冻稀释液中添加大豆卵磷脂对梅花鹿冻融精子质量的影响[J]. 中国畜牧兽医, 2022, 49(11): 4327-4334.
[7]	王宇, 赵予晗, 焦安惠, 王玉琪, 高青山. 乙草胺对小鼠卵母细胞体外成熟的影响[J]. 中国畜牧兽医, 2022, 49(10): 3909-3917.
[8]	王娜, 张洁, 海超, 李欣, 李光鹏, 赵跃芳. 牛精子冷冻损伤及其改善方法[J]. 中国畜牧兽医, 2021, 48(6): 2101-2112.
[9]	王腾飞, 张燕, 王彦平, 赵善江, 朱化彬, 王玲玲, 麻柱, 戴蕴平, 庞云渭. 奶牛活体采卵-体外受精效率的影响因素研究[J]. 中国畜牧兽医, 2021, 48(2): 574-580.
[10]	张雨阳, 汪秋月, 吕艳秋, 陈璇, 徐海峰, 张正文, 金一. 谷氨酰胺替代部分甘油对冻融猪精子获能和体外受精能力的影响[J]. 中国畜牧兽医, 2021, 48(11): 4125-4132.
[11]	艾日夏提·地里夏提, 周正娜, 张也, 刘志强, 章亚峰, 赵茜, 阿布力孜·吾斯曼. 藏红花素对小鼠卵母细胞体外成熟能力的影响[J]. 中国畜牧兽医, 2021, 48(10): 3734-3740.
[12]	菅芯蕊, 钱芙蓉, 王珺, 李晓霞, 张绍萱, 张志彬, 于永生, 高凯, 刁云飞, 张树敏, 李兆华. 聚乙烯吡咯烷酮对公猪精液冷冻的影响[J]. 中国畜牧兽医, 2020, 47(8): 2553-2560.
[13]	吕艳秋, 徐曼, 徐一超, 程咪咪, 金一. 肉碱对冻融猪精子总抗氧化能力、抗氧化酶基因及凋亡相关基因表达的影响[J]. 中国畜牧兽医, 2020, 47(4): 1121-1129.
[14]	努日比娅姆·麦麦提托合提, 赵茜, 艾日夏提·地里夏提, 张博, 阿布力孜·吾斯曼. 白藜芦醇对绵羊卵母细胞体外受精的影响[J]. 中国畜牧兽医, 2020, 47(12): 4000-4007.
[15]	王杰, 李鑫垚, 屈金涛, 张佩华, 雷坤, 詹迎谷, 李莉, 易康乐. 羊腹腔镜子宫角人工输精技术[J]. 中国畜牧兽医, 2019, 46(10): 3016-3022.