曲古抑菌素A对克隆猪端粒长度的影响

doi:10.3724/SP.J.1005.2012.01583

遗传 ›› 2012, Vol. 34 ›› Issue (12): 1583-1590.doi: 10.3724/SP.J.1005.2012.01583

曲古抑菌素A对克隆猪端粒长度的影响

解炳腾¹, 纪光臻², 孔庆然¹, 毛剑², 石永乾¹, 刘世超¹, 武美玲¹, 王娟¹, 刘林², 刘忠华¹

1. 东北农业大学生命科学学院, 哈尔滨150030 2. 南开大学生命科学学院, 天津300192

收稿日期:2012-04-01 修回日期:2012-05-18 出版日期:2012-12-20 发布日期:2012-12-14
通讯作者: 刘忠华 E-mail:liu086@yahoo.com
基金资助:
转基因生物新品种培育科技重大专项：优质转基因猪新品种培育(编号：2008ZX08006-002)和黑龙江省杰出青年基金项目(编号：JC200905)资助

Telomere lengthening by trichostatin A treatment in cloned pigs

XIE Bing-Teng¹, JI Guang-Zhen², KONG Qing-Ran¹, MAO Jian², SHI Yong-Qian¹, LIU Shi-Chao¹, WU Mei-Ling¹, WANG Juan¹, LIU Lin², LIU Zhong-Hua¹

1. Laboratory of Embryo Biotechnology, College of life science, Northeast Agricultural University, Harbin 150030, China 2. Laboratory for Stem Cells and Developmental Biology, College of Life Sciences, Nankai University, Tianjin 300192, China

Received:2012-04-01 Revised:2012-05-18 Online:2012-12-20 Published:2012-12-14

摘要/Abstract

摘要： 端粒是染色体末端结构, 在细胞分裂时随着DNA复制而缩短, 体细胞核移植能不同程度地延长端粒长度, 但有些克隆动物端粒的长度在体细胞核移植过程中不能有效恢复, 因而这些克隆动物就会表现出早衰现象。文章发现克隆东北民猪以及eGFP、Mx和PGC1α转基因克隆猪的端粒长度与核供体成体成纤维细胞相比显著缩短(P<0.05), 表明体细胞核移植的重编程过程没能延长细胞的“寿命”。曲古抑菌素A(Trichostatin A, TSA)是一种去乙酰化酶抑制剂, 有研究表明其能提高某些物种的体细胞核重编程效率。为了使端粒长度有效恢复, 文章利用40 nmol/L TSA处理1细胞期猪克隆胚胎24 h, 结果发现, 与对照组相比, TSA处理能显著地提高克隆胚胎体外发育的囊胚率(16.35% vs. 2 7.09%, 21.60% vs. 34.90%, P<0.05), 而且囊胚期端粒长度也得到显著延长(P<0.05)。克隆胚胎移植受体后得到了TSA处理组与非处理组的克隆猪, 虽然TSA处理并没有提高克隆效率(1.3% vs. 1.7%, TSA vs. control), 但端粒长度与对照组和供体细胞相比均显著延长(P<0.05)。猪体细胞核移植不能有效恢复端粒长度, 但是TSA处理能有效延长克隆猪端粒长度。

关键词: 猪, 克隆, 重编程, 端粒, TSA

Abstract: Telomeres are repeated GC rich sequences at the end of chromosomes, and shorten with each cell division due to DNA end replication problem. Previously, reprogrammed somatic cells of cloned animals display variable telomere elongation. However, it was reported that the cloned animals including Dolly do not reset telomeres and show premature aging. In this study, we investigated telomere function in cloned or transgenic cloned pigs, including the cloned Northeast Min pigs, eGFP, Mx, and PGC1α transgenic cloned pigs, and found that the telomere lengths of cloned pigs were significantly shorter than the nuclear donor adult fibroblasts and age-matched noncloned pigs (P<0.05), indicating that nuclear reprogramming did not restore cellular age of donor cells after somatic cell nuclear transfer (SCNT). Trichostatin A (TSA), an inhibitor of histone deacetylase, has proven to enhance the efficiency of nuclear reprogramming in several species. In order to test whether TSA also can effectively enhance reprogramming of telomeres, TSA (40 nmol/L) was used to treat porcine cloned embryos at 1-cell stage for 24 h. Consistent with previous reports, the developmental rate of SCNT embryos to the blastocyst stage was significantly increased compared with those of the control group (16.35% vs. 27.09%, 21.60% vs. 34.90%, P<0.05). Notably, the telomere length of cloned porcine blastocysts was also significantly elongated (P<0.05). Although TSA did not improve the cloning efficiency (1.3% vs. 1.7%, TSA vs. control), the telomere lengths of cloned piglets were significantly longer compared with those of the control group and the donor fibroblasts (P<0.05). In conclusion, telomeres have not been effectively restored by SCNT in pigs but TSA can effectively lengthen the telomere lengths of cloned pigs.

Key words: pig, cloning, reprogramming, telomere, TSA

解炳腾，纪光臻，孔庆然，毛剑，石永乾，刘世超，武美玲，王娟，刘林，刘忠华. 曲古抑菌素A对克隆猪端粒长度的影响[J]. 遗传, 2012, 34(12): 1583-1590.

XIE Bing-Teng, JI Guang-Zhen, KONG Qing-Ran, MAO Jian SHI Yong-Qian LIU Shi-Chao WU Mei-Ling WANG Juan LIU Lin LIU Zhong-Hua. Telomere lengthening by trichostatin A treatment in cloned pigs[J]. HEREDITAS, 2012, 34(12): 1583-1590.

参考文献

[1] Zakian VA. Telomeres: Beginning to understand the end. Science, 1995, 270(5242): 1601-1607.
[2] Levy MZ, Allsopp RC, Futcher AB, Greider CW, Harley CB. Telomere end-replication problem and cell aging. J Mol Biol, 1992, 225(4): 951-960.
[3] Faragher RGA, Kipling D. How might replicative senescence contribute to human ageing? Bioessays, 1998, 20(12): 985-991.
[4] Shiels PG, Kind AJ, Campbell KHS, Waddington D, Wilmut I, Colman A, Schnieke AE. Analysis of telomere lengths in cloned sheep. Nature, 1999, 399(6734): 316- 317.
[5] Lanza RP, Cibelli JB, Blackwell C, Cristofalo VJ, Francis MK, Baerlocher GM, Mak J, Schertzer M, Chavez EA, Sawyer N, Lansdorp PM, West MD. Extension of cell life-span and telomere length in animals cloned from senescent somatic cells. Science, 2000, 288(5466): 665-669.
[6] Betts DH, Bordignon V, Hill JR, Winger Q, Westhusin ME, Smith LC, King WA. Reprogramming of telomerase activity and rebuilding of telomere length in cloned cattle. Proc Natl Acad Sci USA, 2001, 98(3): 1077-1082.
[7] Miyashita N, Shiga K, Yonai M, Kaneyama K, Kobayashi S, Kojima T, Goto Y, Kishi M, Aso H, Suzuki T, Sakaguchi M, Nagai T. Remarkable differences in telomere lengths among cloned cattle derived from different cell types. Biol Reprod, 2002, 66(6): 1649-1655.
[8] Kishigami S, Bui HT, Wakayama S, Tokunaga K, Van Thuan N, Hikichi T, Mizutani E, Ohta H, Suetsugu R, Sata T, Wakayama T. Successful mouse cloning of an outbred strain by trichostatin a treatment after somatic nuclear transfer. J Reprod Dev, 2007, 53(1): 165-170.
[9] Li J, Svarcova O, Villemoes K, Kragh PM, Schmidt M, Bøgh IB, Zhang Y, Du Y, Lin L, Purup S, Xue Q, Bolund L, Yang H, Maddox-Hyttel P, Vajta G. High in vitro development after somatic cell nuclear transfer and trichostatin a treatment of reconstructed porcine embryos. Theriogenology, 2008, 70(5): 800-808.
[10] Shi LH, Ai JS, Ouyang YC, Huang JC, Lei ZL, Wang Q, Yin S, Han ZM, Sun QY, Chen DY. Trichostatin a and nuclear reprogramming of cloned rabbit embryos. J Anim Sci, 2008, 86(5): 1106-1113.
[11] Takakura M, Kyo S, Sowa Y, Wang Z, Yatabe N, Maida Y, Tanaka M, Inoue M. Telomerase activation by histone deacetylase inhibitor in normal cells. Nucleic Acids Res, 2001, 29(14): 3006-3011.
[12] Khaw AK, Silasudjana M, Banerjee B, Suzuki M, Baskar R, Hande MP. Inhibition of telomerase activity and human telomerase reverse transcriptase gene expression by histone deacetylase inhibitor in human brain cancer cells. Mutat Res, 2007, 625(1-2): 134-144.
[13] 刘忠华, 田江天, 郑重, 王振坤, 宋军, 尹智, 高力, 马海鹍, 孔庆然, 孙爽, 李玉田, 王洪斌. 体细胞核移植克隆民猪, 培养液对卵母细胞成熟及胚胎发育的影响. 中国科学(C辑), 2007, 37(6): 634-640.
[14] Cawthon RM. Telomere measurement by quantitative pcr. Nucleic Acids Res, 2002, 30(10): e47.
[15] 孔庆然, 朱江, 黄波, 郇延军, 王峰, 石永乾, 刘仲凤, 武美玲, 刘忠华. TSA促进转基因猪体细胞核移植胚胎发育和外源基因表达. 遗传, 2011, 33(7): 749-756.
[16] Schaetzlein S, Lucas-Hahn A, Lemme E, Kues WA, Dorsch M, Manns MP, Niemann H, Rudolph KL. Telomere length is reset during early mammalian embryogenesis. Proc Natl Acad Sci USA, 2004, 101(21): 8034-8038.
[17] Liu L, Bailey SM, Okuka M, Munoz P, Li C, Zhou LJ, Wu C, Czerwiec E, Sandler L, Seyfang A, Blasco MA, Keefe DL. Telomere lengthening early in development. Nat Cell Biol, 2007, 9(12): 1436-1441.
[18] Hou M, Wang XB, Popov N, Zhang AJ, Zhao XY, Zhou R, Zetterberg A, Björkholm M, Henriksson M, Gruber A, Xu DW. The histone deacetylase inhibitor trichostatin a derepresses the telomerase reverse transcriptase (hTERT) gene in human cells. Exp Cell Res, 2002, 274(1): 25-34.
[19] Blasco MA. The epigenetic regulation of mammalian telomeres. Nat Rev Genet, 2007, 8(4): 299-309.
[20] Yamanaka K, Sugimura S, Wakai T, Kawahara M, Sato E. Acetylation level of histone h3 in early embryonic stages affects su

编辑推荐

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

曲古抑菌素A对克隆猪端粒长度的影响

Telomere lengthening by trichostatin A treatment in cloned pigs

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	王承贤, 容益康, 崔敏. 果蝇限制端粒转座子的分子机制[J]. 遗传, 2023, 45(3): 221-228.
[2]	田智琛, 尹晓娟. 诱导多能干细胞在儿童疾病的应用研究进展[J]. 遗传, 2023, 45(1): 42-51.
[3]	李亚楠, 张贤君, 张宁, 梁雅琳, 张宇星, 招华兴, 李紫聪, 黄思秀. 过表达组蛋白H3K9me3去甲基化酶对猪克隆胚胎发育的影响[J]. 遗传, 2023, 45(1): 67-77.
[4]	高菲, 王煜, 杜嘉祥, 杜旭光, 赵建国, 潘登科, 吴森, 赵要风. 遗传修饰猪模型在生物医学及农业领域研究进展及应用[J]. 遗传, 2023, 45(1): 6-28.
[5]	崔浩亮, 史佩华, 高锦春, 张新博, 赵顺然, 陶晨雨. 细胞重编程过程中核小体定位改变研究进展[J]. 遗传, 2022, 44(3): 208-215.
[6]	唐湘薇, 楚丹, 颜赛娜, 尹艳飞, 卞桥, 翁波, 陈斌, 冉茂良. miR-191靶向BDNF基因通过激活PI3K/AKT信号通路促进猪未成熟支持细胞增殖[J]. 遗传, 2021, 43(7): 680-693.
[7]	蒋婧, 赵安麒, 谢甜, 陈淑藯, 李劲松. 全基因组蛋白质的标签细胞和小鼠资源库建设[J]. 遗传, 2021, 43(7): 704-714.
[8]	周子文, 王雪, 丁向东. 基于高密度SNP标记估计群体间遗传关联[J]. 遗传, 2021, 43(4): 340-349.
[9]	彭定威, 李瑞强, 曾武, 王敏, 石翾, 曾检华, 刘小红, 陈瑶生, 何祖勇. 编辑MSTN半胱氨酸节基元促进两广小花猪肌肉生长[J]. 遗传, 2021, 43(3): 261-270.
[10]	魏强, 奥岩, 杨漫漫, 陈涛, 韩虎, 张兴举, 王然, 夏秋菊, 姜芳芳, 李勇. 利用全基因组重测序技术鉴定五指山猪GHR突变体转基因插入位点[J]. 遗传, 2021, 43(12): 1149-1158.
[11]	韩程程, 夏凯, 龚茹莹, 吴栩涵, 张蕾, 梁新乐. 适于检测非洲猪瘟病毒的点亮Spinach-p54 RNA适配体的设计及应用[J]. 遗传, 2021, 43(12): 1170-1178.
[12]	邢宝松, 王璟, 陈俊峰, 马强, 任巧玲, 张家庆, 张华, 滑留帅, 孙加节, 曹海. 去势和非去势公猪背最长肌circRNA差异表达分析[J]. 遗传, 2021, 43(11): 1066-1077.
[13]	曹岚, 李志强, 师咏勇, 刘赟. 端粒长度与2型糖尿病：孟德尔随机化研究与多基因风险评分分析[J]. 遗传, 2020, 42(9): 882-888.
[14]	周俊, 赵成成, 吴霄, 石俊松, 周荣, 吴珍芳, 李紫聪. 猪耳成纤维细胞转录组异质性及对核移植胚胎发育的潜在影响[J]. 遗传, 2020, 42(9): 898-915.
[15]	赵利楠, 王娜, 杨国良, 苏现斌, 韩泽广. 基于单细胞靶向测序探究基因碱基突变的方法[J]. 遗传, 2020, 42(7): 703-712.