猪精子获能、顶体反应及冷休克对顶体酶抑制剂的影响
摘要
为了探究猪精子在获能、顶体反应及冷休克处理后对精子顶体酶抑制剂表达量的影响,以及顶体酶抑制剂在精子中何时被降解,低温是否会损伤顶体酶抑制剂,利用SDS-PAGE、Western blot分析射精精子、获能精子、顶体反应精子和冷休克精子组的总蛋白种类及猪精子顶体酶抑制剂表达量的变化。结果显示:射精精子处理组与获能精子、顶体反应精子处理组的顶体酶抑制剂表达量差异显著(P<0.05),且获能和顶体反应处理组间的差异不明显(P>0.05);射精精子与冷休克精子处理组的顶体酶抑制剂表达量差异极显著(P<0.01),且冷休克处理组的顶体酶抑制剂出现异常表达。推测:精子在获能或顶体反应期间,顶体酶抑制剂可能通过泛素-蛋白酶体途径被降解,释放出有活性的顶体酶;低温可能导致顶体酶抑制剂蛋白结构的改变或损坏,导致其表达量异常。
引文
[1]DavidováN,JonákováV,Maňásková-PostlerováP.Expression and localization of acrosin inhibitor in boar reproductive tract[J].Cell Tissue Res,2009(338):303-311.
[2]资捷,宋平,王莉平.NO通过人精子顶体酶对顶体反应的影响[J].中国男科学杂志,2006,20(5):42-43,47.
[3]JelínkováP,MaňáskováP,TicháM,et al.Proteinase inhibitors in aggregated forms of boar seminal plasma proteins[J].Int J Biol Macromol,2003,32(3/4/5):99-107.
[4]de Jonge C.The cAMP-dependent kinase pathway and human sperm acrosomal exocytosis[J].Frontiers in bioscience:a journal and virtual library,1996,1:d234.
[5]Barbelemy C,Royere D,Hammahah S,et al.Ultrastructural changes in membranes and acrosome of human sperm during cryopreservation[J].Arch Aodrol,1990,25(1):29-40.
[6]朱伟杰,姚汝华.冷冻保存对人类精子顶体蛋白酶的影响[J].暨南大学学报,1998,19(3):105-108.
[7]Harrison R A,Flechon J E.Immunocytochemical detection of acrosomal damage following cold shock:loss of acrosin from the acrosomal region of ram,bull and boar spermatozoa[J].Reprod Nutr Dev,1980,20(6):1801-1810.
[8]Jiang J H,Liu X F,Zhen C H,et al.Acrosin structure-based design,synthesis and biological activities of 7-azaindol derivatives as new acrosin inhibitors[J].Chin Chem Lett,2011,22(3):272-275.
[9]蔡萼.精子诱发顶体反应与精子顶体酶活性测定关系的研究[J].中国男科学杂志,2010,24(7):34-35.
[10]Cui Y H,Zhao R L,Wang Q,et al.Determination of sperm acrosin activity for evaluation of male fertility[J].Asian J Androl,2000(2):229-232.
[11]宋桂红.顶体酶抑制剂对精子形态及功能的影响[D].上海:第二军医大学,2008.
[12]邵毓敏.泛素—蛋白酶体通路在生精过程中的作用[D].北京:中国协和医科大学,2006.
[13]Nayernia K,Adham I M,Shamsadin R,et al.Proacrosin-deficient mice and zona pellucida modification in an expermental model of multifactorial infertility[J].Mol Hum Reprod,2002,8(5):434-440.
[14]Yi Y J,Manandhar G,Sutovsky M,et al.Inhibition of 19S proteasomal regulatory complex subunit PSMD8 increases polyspermy during porcine fertilization in vitro[J].Reprod Immunol,2010,84(2):154-163.
[15]朴海仙,金一,韩明铭.温度和保存时间对猪精子顶体酶活性的影响[J].安徽农业大学学报,2010,37(1):54-57.
[16]Aitken R J.Free radicals,1ipid peroxidation and sperm function[J].Reprod Fert Develop,1995,7(4):659-668.
[2]资捷,宋平,王莉平.NO通过人精子顶体酶对顶体反应的影响[J].中国男科学杂志,2006,20(5):42-43,47.
[3]JelínkováP,MaňáskováP,TicháM,et al.Proteinase inhibitors in aggregated forms of boar seminal plasma proteins[J].Int J Biol Macromol,2003,32(3/4/5):99-107.
[4]de Jonge C.The cAMP-dependent kinase pathway and human sperm acrosomal exocytosis[J].Frontiers in bioscience:a journal and virtual library,1996,1:d234.
[5]Barbelemy C,Royere D,Hammahah S,et al.Ultrastructural changes in membranes and acrosome of human sperm during cryopreservation[J].Arch Aodrol,1990,25(1):29-40.
[6]朱伟杰,姚汝华.冷冻保存对人类精子顶体蛋白酶的影响[J].暨南大学学报,1998,19(3):105-108.
[7]Harrison R A,Flechon J E.Immunocytochemical detection of acrosomal damage following cold shock:loss of acrosin from the acrosomal region of ram,bull and boar spermatozoa[J].Reprod Nutr Dev,1980,20(6):1801-1810.
[8]Jiang J H,Liu X F,Zhen C H,et al.Acrosin structure-based design,synthesis and biological activities of 7-azaindol derivatives as new acrosin inhibitors[J].Chin Chem Lett,2011,22(3):272-275.
[9]蔡萼.精子诱发顶体反应与精子顶体酶活性测定关系的研究[J].中国男科学杂志,2010,24(7):34-35.
[10]Cui Y H,Zhao R L,Wang Q,et al.Determination of sperm acrosin activity for evaluation of male fertility[J].Asian J Androl,2000(2):229-232.
[11]宋桂红.顶体酶抑制剂对精子形态及功能的影响[D].上海:第二军医大学,2008.
[12]邵毓敏.泛素—蛋白酶体通路在生精过程中的作用[D].北京:中国协和医科大学,2006.
[13]Nayernia K,Adham I M,Shamsadin R,et al.Proacrosin-deficient mice and zona pellucida modification in an expermental model of multifactorial infertility[J].Mol Hum Reprod,2002,8(5):434-440.
[14]Yi Y J,Manandhar G,Sutovsky M,et al.Inhibition of 19S proteasomal regulatory complex subunit PSMD8 increases polyspermy during porcine fertilization in vitro[J].Reprod Immunol,2010,84(2):154-163.
[15]朴海仙,金一,韩明铭.温度和保存时间对猪精子顶体酶活性的影响[J].安徽农业大学学报,2010,37(1):54-57.
[16]Aitken R J.Free radicals,1ipid peroxidation and sperm function[J].Reprod Fert Develop,1995,7(4):659-668.