红海海绵素A对小鼠精子冷冻活力的影响

doi:10.3969/j.issn.1674-5817.2013.01.002

实验动物与比较医学 ›› 2013, Vol. 33 ›› Issue (1): 7-11.DOI: 10.3969/j.issn.1674-5817.2013.01.002

红海海绵素A对小鼠精子冷冻活力的影响

程芙蓉¹, 杨世华²

1.深圳市罗湖区人民医院,深圳518001;
2.昆明理工大学生命科学与技术学院, 昆明650500

收稿日期:2012-06-07 出版日期:2013-02-25 发布日期:2013-02-25
作者简介:程芙蓉(1982-), 女, 博士, 从事传染性疾病与生殖疾病控制研究, E-mail: cheng820421@163.com
基金资助:
国家自然科学基金项目(31071279); 云南省科技创新人才计划项目(2011CI009)

Effect of Latrunculin A on Motility of Spermatozoa Cryopreservation in ICR Mouse

CHENG Fu-rong¹, YANG Shi-hua²

1. Shenzhen Louhu People’s Hospital, Shenzhen 518001, China;
2. Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650093, China

Received:2012-06-07 Online:2013-02-25 Published:2013-02-25

摘要/Abstract

摘要： 目的通过建立最佳的冷冻降温速率,研究红海海绵素A(latrunculin A, LATA)对精子的冷冻保护作用。方法用冷冻液平衡后的ICR小鼠附睾精子置于液氮面上方不同的位置,用快速 (-240℃/min)、中速(-109℃/min)、慢速(-41℃/min)和超慢速(-12℃/min)的降温速率分别冷冻5 或10 min后投入液氮冻存。结果在液氮上方冷冻5 min或10 min对小鼠精子的复苏活力没有影响。而用超慢速冷冻速率冻存精子的复苏活力显著高于其他组,中速冷冻速率保存的精子的复苏活力在各实验组中最低。用0.5 µmol/L、1 µmol/L、2 µmol/L和4 µmol/L 的红海海绵素A处理小鼠精子后,再用超慢速冷冻速率保存,结果用0.5 µmol/L、1 µmol/L和2 µmol/L 的红海海绵素A处理的精子未能提高精子的复苏活力,而高浓度的红海海绵素A(4 µmol/L)反而降低了精子的复苏活力度。结论红海海绵素A对精子的冷冻保护作用值得商榷。

关键词: 小鼠精子, 冷冻降温速率, 红海海绵素A, 活力

Abstract: Objective To evaluate the effect of latrunculin A on motility of spermatozoa combined with optimized cooling rate of cryopreservation. Methods ICR mouse epididymal spermatozoa balanced in freezing medium were frozen at fast (-240℃/min), medium (-109℃/min), slow (-41℃/min) and super slow (-12℃/min) cooling rate for 5 min or 10 min respectively, before being plunged into liquid nitrogen for storage. Results No difference of sperm motility was observed between spermatozoa frozen for 5 and 10 min above liquid nitrogen at each cooling rate. Spermatozoa cryopreserved at super slow and medium cooling rate showed highest and lowest post-thaw motility respectively. ICR mouse spermatozoa were treated with 0.5 µmol/L, 1 µmol/L, 2 µmol/L and 4 µmol/L Latrunculin A before freezing and then cryopreserved at super slow cooling rate. The result showed that treating mouse spermatozoa with 0.5 µmol/L, 1 µmol/L and 2 µmol/L of Latrunculin A could not improve the motility of spermatozoa. However, high level (4 µmol/L) of Latrunculin A showed detrimental effect on ICR mouse sperm motility. Conclusion Cryopreservation proficiency of Latrunculin A on sperm is open to discussion.

Key words: Mouse spermatozoa, Cooling rate, Latrunculin A, Motility

中图分类号:

Q95-33

程芙蓉, 杨世华. 红海海绵素A对小鼠精子冷冻活力的影响[J]. 实验动物与比较医学, 2013, 33(1): 7-11.

CHENG Fu-rong, YANG Shi-hua. Effect of Latrunculin A on Motility of Spermatozoa Cryopreservation in ICR Mouse[J]. Laboratory Animal and Comparative Medicine, 2013, 33(1): 7-11.

参考文献

[1] Waterston R, Lindblad-Toh K, Birney E, et al.Initial sequencing and comparative analysis of the mouse genome[J]. Nature, 2002, 420:520-562.
[2] Green JE, Hudson T.The promise of genetically engineered mice for cancer prevention studies[J]. Nat Rev Cancer, 2005, 5:184-198.
[3] Seong E, Saunders TL, Stewart CL, et al.To knockout in 129 or in C57BL/6: that is the question[J]. Trends Genet, 2004, 20:59-62.
[4] Thornton C, Brown SD, Glenister PH.Large numbers of mice established by in vitro fertilization with cryopreserved spermatozoa: implications and applications for genetic resource banks, mutagenesis screens, and mouse backcrosses[J]. Mamm Genome, 1999, 10:987-992.
[5] Nakagata N.Cryopreservation of mouse spermatozoa[J]. Mamm Genome, 2000, 11:572-576.
[6] Mazur P.Cryobiology: the freezing of biological systems[J]. Science, 1970, 168:939-949.
[7] Stacy R, Eroglu A, Fowler A, et al.Thermal characterization of Nakagata’s mouse sperm freezing protocol[J]. Cryobiology, 2006, 52:99-107.
[8] Koshimoto C, Mazur P.Effects of cooling and warming rate to and from -70 degrees C, and effect of further cooling from -70 to -196 degrees C on the motility of mouse spermatozoa[J]. Biol Reprod, 2002, 66:1477-1484.
[9] Sztein JM, Farley JS, Mobraaten LE.In vitro fertilization with cryopreserved inbred mouse sperm[J]. Biol Reprod, 2000, 63:1774-1780.
[10] Walters EM, Men H, Agca Y, et al.Osmotic tolerance of mouse spermatozoa from various genetic backgrounds: acrosome integrity, membrane integrity, and maintenance of motility[J]. Cryobiology, 2005, 50:193-205.
[11] Willoughby CE, Mazur P, Peter AT, et al.Osmotic tolerance limits and properties of murine spermatozoa[J]. Biol Reprod, 1996, 55:715-727.
[12] Mazur P, Leibo SP, Chu EH.A two-factor hypothesis of freezing injury. Evidence from Chinese hamster tissue-culture cells[J]. Exp Cell Res, 1972, 71:345-355.
[13] Agca Y, Mullen S, Liu J, et al.Osmotic tolerance and membrane permeability characteristics of rhesus monkey (Macaca mulatta) spermatozoa[J].Cryobiology, 2005, 51:1-14.
[14] Alapati R, Goff K, Kubisch HM, et al.Water transport in epididymal and ejaculated rhesus monkey (Macaca mulatta) sperm during freezing[J]. Cryobiology, 2008, 57:182-185.
[15] Devireddy RV, Fahrig B, Godke RA, et al.Subzero water transport characteristics of boar spermatozoa confirm observed optimal cooling rates[J]. Mol Reprod Dev, 2004, 67:446-457.
[16] Gilmore JA, Liu J, Peter AT, et al.Determination of plasma membrane characteristics of boar spermatozoa and their relevance to cryopreservation[J]. Biol Reprod, 1998, 58:28-36.
[17] Gilmore JA, McGann LE, Liu J, et al. Effect of cryoprotectant solutes on water permeability of human spermatozoa[J]. Biol Reprod, 1995,53:985-995.
[18] Mazur P.Limits to life at low temperatures and at reduced water contents and water activities[J]. Orig Life, 1980, 10:137-159.
[19] Eroglu A, Toth TL, Toner M.Alterations of the cytoskeleton and polyploidy induced by cryopreservation of metaphase II mouse oocytes[J]. Fertil Steril, 1998, 69:944-957.
[20] Morton WM, Ayscough KR, McLaughlin PJ. Latrunculin alters the actin-monomer subunit interface to prevent polymerization[J]. Nature Cell Biol, 2000, 2:376-378.
[21] Hosu BG, Mullen SF, Critser JK, Forgacs G.Reversible disassembly of the actin cytoskeleton improves the survival rate and developmental competence of cryopreserved mouse oocytes[J]. PLoS ONE, 2008, 3:e2787.
[22] Agca Y, Gilmore J, Byers M, Woods EJ, Liu J, Critser JK.Osmotic characteristics of mouse spermatozoa in the presence of extenders and sugars[J]. Biol Reprod, 2002, 67:1493-1501.

红海海绵素A对小鼠精子冷冻活力的影响

Effect of Latrunculin A on Motility of Spermatozoa Cryopreservation in ICR Mouse

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价

[1]	王秋静, 宋祥福, 李天舒. 兔“有机磷酸酯类中毒及解救”实验中乙酰胆碱酯酶活力测定技术的改进[J]. 实验动物与比较医学, 2020, 40(6): 528-530.
[2]	王美珊, 孔彭成, 朱燕, 朱莲, 陈学进, 李和平, 蒋满喜. 一种改良的小鼠附睾尾常温运输方法[J]. 实验动物与比较医学, 2014, 34(2): 145-148.