超低温冷冻对脊尾白虾精子几种酶活性的影响
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南京农业大学资源与环境科学学院 江苏省海洋生物学重点实验室,淮海工学院 江苏省海洋生物技术重点建设实验室 江苏 连云港,淮海工学院 江苏省海洋生物技术重点建设实验室 江苏 连云港,南京农业大学资源与环境科学学院 江苏省海洋生物学重点实验室,淮海工学院 江苏省海洋生物技术重点建设实验室 江苏 连云港

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“十二五”国家科技支撑计划项目(2011BAD13B03);国家自然科学基金项目(31072213);江苏省水产三项工程项目(P2009-18)


Effects of cryopreservation on enzyme activity of Exopalaemon carinicauda spermatozoa
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College of Resources and Environmental Sciences,Nanjing Agricultural University,China Jiangsu Key Laboratory of Marine Biotechnology,Huaihai Institute of Technology,China Jiangsu Key Laboratory of Marine Biotechnology,Huaihai Institute of Technology,College of Resources and Environmental Sciences,Nanjing Agricultural University,China Jiangsu Key Laboratory of Marine Biotechnology,Huaihai Institute of Technology

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    摘要:

    研究了超低温冷冻保存(-196 ℃)对脊尾白虾精子内琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、Na+/K+-ATP酶、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)与顶体酶活性的影响,以期为提高脊尾白虾精子超低温冷冻效果提供理论依据。设置对照组(未添加抗冻剂)、3个实验组[分别添加DMSO(V/V)10.0%、12.5%、15.0%],于冷冻0、1、3、5、7、15 d 取样测定各酶活性。结果表明,经冷冻后,除GR外,其他所测酶活性均出现显著下降(P<0.05),且以对照组酶活性下降幅度最大。GR活性在冷冻7 d内显著升高,且对照组明显高于实验组(P<0.05),在冷冻15 d时又出现下降。添加15.0%DMSO组所测酶活性均高于同期其它各组,表明15.0%DMSO对精子内酶的保护作用较好。冷冻15 d后15.0%DMSO组的SDH、LDH和Na+/K+-ATP酶活性由(28.500±1.453)U/mL、(1290.836±27.603)U/L和(2.605±0.232)μmol/(mg?h)分别降至(15.300±0.950)U/mL、 (363.713±13.943)U/L和(0.542±0.186)μmol/(mg?h);SOD和CAT活性由(106.497±7.217)U/mL、(383.632±4.731)U/g分别降至(17.036±0.321)U/mL、(166.940±1.910)U/g;顶体酶活性从(3.521±0.010)μIU/106降至(1.212±0.043)μIU/106;而GR活性由(217.042±6.962)U/L上升至(302.787±24.558) U/L。从冷冻后各酶下降幅度来看,超低温冷冻对SOD活性的影响最大,其次是Na+/K+-ATP酶。

    Abstract:

    Exopalaemon carinicauda is one of the marine aquaculture species with high economic value in China.This paper studied the effects of cryopreservation(-196 ℃)on the enzyme activity(SDH,LDH,Na+K+-ATP,SOD,CAT,GR and Acrosin)in E.carinicauda spermatozoa,in order to provide a theoretical basis for cryopreservation effect on E.carinicauda spermatozoa,and lay a foundation for artificial propagation of E.carinicauda.We set up a control group(without cryoprotectant),and three experimental groups(DMSO(V/V)10%,12.5%,15%),detected the enzyme activities on day 0,1,3,5,7,15.The result showed that,after cryopreservation,in addition to GR,the other enzyme activities decreased significantly(P<0.05),with the biggest drop in the control group.GR activity significantly increased in 7 days,and the control group was significantly higher than the experimental group(P<0.05),declining again in 15 days.The group of 15% DMSO measured enzyme activities were higher than other groups,indicating that 15% DMSO had protective effect on enzyme of spermatozoa.15 days later,under the condition of 15% DMSO,the activities of SDH,LDH and Na+/K+-ATPase decreased from(28.500±1.453)U/mL,(1290.836±27.603)U/L and(2.605±0.232)μmol/(mg?h)to(15.300±0.950)U/mL,(363.713±13.943)U/L and(0.542±0.186)μmol/(mg?h)respectively;the activities of SOD and CAT decreased from(106.497±7.217)U/mL,(383.632±4.731)U/g to(17.036±0.321)U/mL and(166.940±1.910)U/g respectively;the activities of acrosin decreased from(3.521±0.010)μIU/106 to(1.212±0.043)μIU/106;but the activities of GR increased from(217.042±6.962)U/L to(302.787±24.558)U/L.From the decline range,cryopreservation had the biggest influence on SOD,followed by Na+/K+-ATPase.

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浦蕴惠,许星鸿,高焕,刘兆普,阎斌伦.超低温冷冻对脊尾白虾精子几种酶活性的影响[J].水产学报,2013,37(1):101~108

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  • 收稿日期:2012-02-22
  • 最后修改日期:2012-10-22
  • 录用日期:2012-12-02
  • 在线发布日期: 2013-01-15
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