程序化冷冻前后小鼠休眠胚胎中Apol8蛋白分布和表达

doi:10.11924/j.issn.1000-6850.casb17030120

中国农学通报 ›› 2017, Vol. 33 ›› Issue (21): 149-153.doi: 10.11924/j.issn.1000-6850.casb17030120

所属专题：生物技术；畜牧兽医

程序化冷冻前后小鼠休眠胚胎中Apol8蛋白分布和表达

王力红,郭勇,翟椿东,卢文瑾,刘迪,杨健,王相国,盛熙晖,齐晓龙,倪和民

(北京农学院动物育种与辅助生殖实验室,北京 102206）

收稿日期:2017-03-20 修回日期:2017-07-10 接受日期:2017-05-01 出版日期:2017-07-27 发布日期:2017-07-27
通讯作者: 王力红
基金资助:
2013 年度北京市教委北京市属高等学校创新团队建设与教师职业发展计划项目“体细胞转基因克隆肉牛新品系培育与利用”(PXM2013_ 014207_000067)；奶牛产业技术体系北京市创新团队。

Distribution and Differential Expression of Apol8 in Mouse Normal Hatched Blastocysts and Dormant Embryos Before and After Cryopreservation

Wang Lihong, Guo Yong, Zhai Chundong, Lu Wenjin, Liu Di, Yang Jian, Wang Xiangguo,Sheng Xihui, Qi Xiaolong, Ni Hemin

(Animal Breeding and Assisted Reproductive Laboratory, Beijing University of Agriculture, Beijing 102206)

Received:2017-03-20 Revised:2017-07-10 Accepted:2017-05-01 Online:2017-07-27 Published:2017-07-27

摘要/Abstract

摘要： 旨在研究小鼠孵化囊胚和休眠胚胎程序化冷冻前、后Apol8蛋白定位和相对表达量变化。通过超排技术获取小鼠体内孵化囊胚，通过小鼠延迟着床模型获取休眠胚胎，通过Confocal显微镜和Western Blot技术对孵化囊胚和休眠胚胎程序化冷冻前后Apol8蛋白的分布和表达进行检测。结果表明：作为抗冻潜力蛋白，在程序化冷冻前、后孵化囊胚和休眠胚胎Apol8均有表达。程序化冷冻后孵化囊胚中Apol8的表达量是冷冻前1.2倍(P＜0.01)，程序化冷冻后休眠囊胚中Apol8的表达量是冷冻前2.0倍(P＜0.01)，程序化冷冻后休眠胚胎Apol8表达量是冷冻后孵化囊胚的1.25倍(P＜0.01)。程序化冷冻后休眠胚胎Apol8相对表达量变化明显大于孵化囊胚，Apol8蛋白具有作为新型哺乳动物源抗冻蛋白的潜力。

关键词: 澳洲坚果, 澳洲坚果, 果实生长, 落果率, 干热河谷

Abstract: This study aimed to investigate the distribution and differential expression of Apol8 protein in both mouse normal hatched blastocysts and dormant embryos before and after cryopreservation. The normal hatched blastocysts were obtained by superovulation, and the dormant embryos were collected from mice with delayed implantation. Confocal microscopy and western blot were applied to detect the distribution and differential expression of Apol8 among different groups. The results showed that Apol8, as an antifreeze protein, was expressed in mouse normal hatched blastocysts and dormant embryos before and after cryopreservation. The expression of Apol8 in normal hatched blastocysts after cryopreservation was 1.2 times that before cryopreservation (P＜0.01). The expression of Apol8 in dormant embryo after cryopreservation was 2.0 times that before cryopreservation (P＜0.01). The expression of Apol8 in dormant embryo after cryopreservation was 1.25 times that in normal hatched blastocyst after cryopreservation (P＜0.01). The expression level of Apol8 in dormant embryo was significantly higher than that of normal hatched blastocysts. Apol8 has the potential to be used as a novel mammalian-derived antifreeze protein.

中图分类号:

S814.8

王力红,郭勇,翟椿东,卢文瑾,刘迪,杨健,王相国,盛熙晖,齐晓龙,倪和民. 程序化冷冻前后小鼠休眠胚胎中Apol8蛋白分布和表达[J]. 中国农学通报, 2017, 33(21): 149-153.

Wang Lihong,Guo Yong,Zhai Chundong,Lu Wenjin,Liu Di,Yang Jian,Wang Xiangguo,Sheng Xihui,Qi Xiaolong and Ni Hemin. Distribution and Differential Expression of Apol8 in Mouse Normal Hatched Blastocysts and Dormant Embryos Before and After Cryopreservation[J]. Chinese Agricultural Science Bulletin, 2017, 33(21): 149-153.

参考文献

[1] Whittingham D G. Survival of mouse embryos after freezing and thawing[J].Nature,1971,233:125-126.
[2] Wilmut I, Rowson L E. Experiments on the low-temperature preservation of cow embryos[J].Vet Rec,1973,92:686-690.
[3] Willadsen S M, Polge C, Rowson L E, et al. Deep freezing of sheep embryos[J].J Reprod Fertil,1976,46:151-154.
[4] 谢秀杰,贾宗超,魏群.抗冻蛋白结构与抗冻机制[J].细胞生物学杂志,2005(27):5-8.
[5] 姚佳.抗冻蛋白及其抗冻机理研究进展[J].四川畜牧兽医,2013(4):30-32.
[6] 樊绍刚,张党权,邓顺阳,等.抗冻蛋白和冰核蛋白对植物抗冻性能的作用机制[J].经济林研究,2009,27(2):125-130.
[7] 汪少芸,李晓坤,周焱富,等.抗冻蛋白的作用机制及基因工程研究进展[J].北京工商大学学报,2012,30( 2):58-62.
[8] Yu S O, Brown A, Iddleton A J, et al. Ice restructuring inhibition activities in antifreeze proteins with distinct differences in thermal hysteresis[J].Cryobiology,2010,61:327-334.
[9] 费云标,魏令波,高素琴.添加植物抗冻蛋白提高猪胚胎冷冻保存成活率的初步试验[J].黑龙江动物繁殖,1995,3(2):21-23.
[10] Lim J M, Hansel W. Improved development of in vitro-derived bovine embryos by use of a nitric oxide scavenger in a cumulusgranulosa cell coculture system[J].Mol Reprod,1998,50:45-43.
[11] Lim J M, Mei Y, Chen B, et al. Development of bovine IVF oocytes cultured in medium supplemented with a nitric oxide scavenger or inhibitor in a coculture system[J].Theriogenology,1999,51:941-949.
[12] Achache H, Revel A. Endometrial receptivity markers, the journey to successful embryo implantation[J].Hum Reprod Update,2006,12:731-746.
[13] Ruud J T. Vertebrates without erythrocytes and blood pigment[J].Nature,1954,173(4410):84-850.
[14] Lopes F L, Desmarais J A, Murphy B D. Embryonic diapause and its regulation[J].Reproduction,2004,128(6):669-678.
[15] Johnson D, Chatterjee S. Embryo implantation in the rat uterus induced by epidermal growth factor[J].Rreproduction and fertility,1993,99(2):557-559.
[16] Petak G E, Taccoin E, Czemik, et al. Embryonic diapause is conserved across mammals[J].PLoS One,2012,7(3):1-7.
[17] 张劭俣,刘云海,倪和民,等.经程序化冷冻的小鼠休眠胚胎的基因表达谱差异分析[J].中国实验动物学报,2012,20(5):15-20.
[18] 顾美超,卢天罡,刘云海,等.冻融后小鼠休眠胚胎超微结构变化的研究[J].中国实验动物学报,2014,22(3):53-56.
[19] Paria B, Das S K, Andrews G K, et al. Expression of the epidermal growth factor receptor gene is regulated in mouse blastocysts during delayed implantation[J].Proc Nat Acad Sci,1993,90(1):55-59.
[20] Paria B, Huet-Hudson Y, Dey S. Blastocystimes state of activity determines the “window” of implantation in the receptive mouse uterus[J].Proc Nat Acad Sci,1993,90(21):10159-10162.

程序化冷冻前后小鼠休眠胚胎中Apol8蛋白分布和表达

Distribution and Differential Expression of Apol8 in Mouse Normal Hatched Blastocysts and Dormant Embryos Before and After Cryopreservation

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