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

• 研究报告 • 上一篇    下一篇

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. 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. 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: 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