低温和获能对猪精子细胞骨架微管蛋白和肌动蛋白表达量的影响
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摘要
【目的】探究获能和低温冷冻对猪精子细胞骨架中微管蛋白和肌动蛋白表达量的影响。【方法】利用SDS-PAGE、Western blot技术,分析获能和低温冷冻(-80和-196℃)后猪精子总蛋白质种类以及猪精子细胞骨架微管蛋白和肌动蛋白表达量的变化。【结果】低温处理后,猪精子总蛋白质的种类与新鲜精子相比发生变化,而获能处理后几乎没有差异。低温冷冻导致猪精子细胞骨架微管蛋白和肌动蛋白的表达量较新鲜精子下降,且-196℃冷冻组肌动蛋白的表达量要略高于-80℃冷冻组,但二者微管蛋白的表达量基本没有差异。获能处理后猪精子肌动蛋白表达量略有下降,而微管蛋白表达量基本没有变化。【结论】低温冷冻处理对猪精子总蛋白质的种类以及猪精子细胞骨架微管蛋白和肌动蛋白的表达量有明显影响,而获能处理则影响较小。
【Objective】 This study aimed to examine the effects of capacitation and low temperature on expression of tubulin and actin in boar sperm.【Method】 Changes of tubulin and actin expression and types of total sperm protein were analyzed by SDS-PAGE and Western bolt after capacitation and low temperature treatments were conducted in this experiment.【Result】 Types of total sperm proteins changed significantly after treatment of low temperature,while capacitation did not change the proteins.The expression level of cytoskeletal tubulin and actin decreased after treatment of low temperature.Expression of tubulin after being frozen in-196 ℃ was higher than being frozen in-80 ℃,while expression of tubulin was almost same at these two temperatures.【Conclusion】 Low temperature treatments affect significantly the types of total sperm proteins and expression of cytoskeletal tubulin and actin,but effects of capacitation are neglectable.
【Objective】 This study aimed to examine the effects of capacitation and low temperature on expression of tubulin and actin in boar sperm.【Method】 Changes of tubulin and actin expression and types of total sperm protein were analyzed by SDS-PAGE and Western bolt after capacitation and low temperature treatments were conducted in this experiment.【Result】 Types of total sperm proteins changed significantly after treatment of low temperature,while capacitation did not change the proteins.The expression level of cytoskeletal tubulin and actin decreased after treatment of low temperature.Expression of tubulin after being frozen in-196 ℃ was higher than being frozen in-80 ℃,while expression of tubulin was almost same at these two temperatures.【Conclusion】 Low temperature treatments affect significantly the types of total sperm proteins and expression of cytoskeletal tubulin and actin,but effects of capacitation are neglectable.
引文
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[13]韩明铭,金一,方南洙,等.获能和冻融猪精子间细胞肌动蛋白表达的差异[J].西北农林科技大学学报:自然科学版,2007,37(11):47-50.Han M M,Jin Y,Fang N Z,et al.Difference between capaci-tation and frozen-thawed cell actin expression in boar sperm[J].Journal of Northwest A&F University:Nat Sci Ed,2007,37(11):47-50.(in Chinese)
[14]Cabello-Agueros J F,Hernandez-Gonzalez E O,Mujica A.Therole of F-actin cytoskeleton-associated gelsolin in the guineapig capacitation and acrosome reaction[J].Cell Motility andthe Cytoske-leton,2003,56(2):94-108.
[15]Rogers B J,Bastias C,Coulson R L,et al.Cytochalasin D inhibits penetration of hamster eggs by guinea pig and humanspermatozoa[J].Journal ofAndrology,1989,10(4):275-282.
[16]Brener E,Rubinstein S,Cohen G,et al.Remodeling of the ac-tin cytoskeleton during mammalian sperm capacitation and ac-rosome reaction[J].Biology of Reproduction,2003,68(4):837-845.
[2]Chen C K,Wang C W,Tsai W J,et al.Evaluation of meioticspindles in thawed oocytes after vitrification using polarizedlight microscopy[J].FertiSteril,2004,82(3):666-672.
[3]Kim N H,Day B N,Lee H T,et al.Mierofilament assemblyand cortical granule distribution during maturation,partheno-genetic activation and fertilization in the porcine oocyte[J].Zygote,1996,4(2):145-149.
[4]熊培颖,高黄生.细胞骨架与力信号传导[J].国外医学口腔医学分册,2005,32(2):120-122.Xiong P Y,Gao H S.Cytoskeleton and force signal transduc-tion[J].Foreign Medical Sciences,2005,33(2):120-122.(inChinese)
[5]袁韦军.细胞骨架的基本成分与功能[J].生物学教学,2006,32(4):5-8.Yuan W J.The basic components and functions of the cytoskel-eton[J].Biology Teaching,2006,32(4):5-8.(in Chinese)
[6]夏其昌.蛋白质电泳技术指南[M].北京:化学工业出版社,2007.Xia Q C.Protein electrophoresis technical guide[M].Beijing:Chemical Industry Press,2007.(in Chinese)
[7]Frixione E.Recurring views on the structure and function ofthe cytoskeleton[J].Cell Motility and the Cytoseleton,2000,46(2):73-94.
[8]金伯泉.细胞和分子免疫学[M].北京:科技出版社,2007:388-391.Jin B Q.Cellular and molecular immunology[M].Beijing:Sci-ence and Technology Press,2007:388-391.(in Chinese)
[9]Katerina D,Harry D M,Natasa Sebkova,et al.Cytoskeletonlocalization in the sperm head prior to fertilization[J].Repro-duction,2005,130(3):61-69.
[10]Dvorakova K,Palecek J,Peknicova J.Changes in immuno-cy-tochemical localization of cytoskeletal proteins in boar sper-matozoa after the AR induced by specific cytoskeletal inhibi-tors[J].Folia Biologica,2001,47(2):18-27.
[11]Liu R H,Sun Q Y,Li Y H,et al.Effects of cooling on meioticspindle structure and chromosome alignment within in vitromatured porcine oocytes[J].Molecular Reproduction and De-velopment,2003,65(1):212-218.
[12]武彩红,芮荣,戴建军,等.猪卵母细胞玻璃化冷冻后细胞骨架的变化[J].动物学研究,2006,27(4):382-389.Wu C H,Rui R,Dai J J,et al.Changes of the cytoskeleton incell vitrification of pig oocytes[J].Zoological Research,2006,27(4):382-389.(in Chinese)
[13]韩明铭,金一,方南洙,等.获能和冻融猪精子间细胞肌动蛋白表达的差异[J].西北农林科技大学学报:自然科学版,2007,37(11):47-50.Han M M,Jin Y,Fang N Z,et al.Difference between capaci-tation and frozen-thawed cell actin expression in boar sperm[J].Journal of Northwest A&F University:Nat Sci Ed,2007,37(11):47-50.(in Chinese)
[14]Cabello-Agueros J F,Hernandez-Gonzalez E O,Mujica A.Therole of F-actin cytoskeleton-associated gelsolin in the guineapig capacitation and acrosome reaction[J].Cell Motility andthe Cytoske-leton,2003,56(2):94-108.
[15]Rogers B J,Bastias C,Coulson R L,et al.Cytochalasin D inhibits penetration of hamster eggs by guinea pig and humanspermatozoa[J].Journal ofAndrology,1989,10(4):275-282.
[16]Brener E,Rubinstein S,Cohen G,et al.Remodeling of the ac-tin cytoskeleton during mammalian sperm capacitation and ac-rosome reaction[J].Biology of Reproduction,2003,68(4):837-845.