In vivo and in vitro development of Tibetan antelope (Pantholops hodgsonii) interspecific cloned embryos

Abstract

The Tibetan antelope is endemic to the Tibetan Plateau, China, and is now considered an endangered species. As a possible rescue strategy, the development of embryos constructed by interspecies somatic cell nuclear transfer (iSCNT) was examined. Tibetan antelope fibroblast cells were transferred into enucleated bovine, ovine and caprine oocytes. These cloned embryos were then cultured in vitro or in the oviducts of intermediate animals.

Less than 0.5% of the reconstructed antelope-bovine embryos cultured in vitro developed to the blastocyst stage. However, when the cloned antelope-bovine embryos were transferred to caprine oviducts, about 1.6% of the embryos developed to the blastocyst stage. In contrast, only 0.7% of the antelope-ovine embryos developed to the morula stage and none developed to blastocysts in ovine oviducts. The treatment of donor cells and bovine oocytes with trichostatin A did not improve the embryo development even when cultured in the oviducts of ovine and caprine. When the antelope-bovine embryos, constructed from oocytes treated with roscovitine or trichostatin A, were cultured in rabbit oviducts 2.3% and 14.3% developed to blastocysts, respectively. It is concluded that although some success was achieved with the protocols used, interspecies cloning of Tibetan antelope presents difficulties still to be overcome. The mechanisms resulting in the low embryo development need investigation and progress might require a deeper understanding of cellular reprogramming.

Keywords

interspecific nuclear transfer / bovine / ovine / caprine / oviduct / apoptosis

References

[1] Lei R H, Jian Z G, Hu Z, Yang W. Phylogenetic relationships of Chinese antelopes (sub family Antilopinae) based on mitochondrial ribosomal RNA gene sequences. Journal of Zoology, 2003, 261(3): 227-237

[2] Ruan X D, He P J, Zhang J L, Wan Q H, Fang S G. Evolutionary history and current population relationships of the chiru (Pantholops hodgsonii) inferred from mtDNA variation. Journal of Mammalogy, 2005, 86(5): 881-886

[3] Leslie D M, Schaller G B. Pantholops hodgsonii (Artiodactyla: Bovidae). Mammalian Species, 2008, 817(1): 1-13

[4] Mallon D P. Pantholops hodgsonii. In: IUCN 2008. IUCN Red List of Threatened Species, 2008

[5] Fei J, Yang J, Zhou H, Tang M, Lu W, Yan A, Hou Y, Zhang S. A novel method for identifying shahtoosh. Journal of Forensic Sciences, 2014. first published online〕

[6] Chen D, Sun Q, Liu J, Li G, Lian L, Wang M, Han Z, Song X, Li J, Sun Q, Chen Y, Zhang Y, Ding B. The giant panda (Ailuropoda melanoleuca) somatic nucleus can dedifferentiate in rabbit ooplasm and support early development of the reconstructed egg. Science in China (Series C), 1999, 42(4): 346-353

[7] Li J S, Chen D Y, Han Z M, Zhu Z Y, Wen R C, Liu Z H, Wang M K, Lian L, Du J, Wang P Y, Zhang H M. The effect of series nuclear transfer protocol on the embryonic development of the giant panda interpecies cloning. Chinese Science Bulletin, 2001, 46(22): 1899-1901

[8] Zhao Z J, Li R C, Cao H H, Zhang Q J, Jiang M X, Ouyang Y C, Nan C L, Lei Z L, Song X F, Sun Q Y, Chen D Y. Interspecies nuclear transfer of Tibetan antelope using caprine oocyte as recipient. Molecular Reproduction and Development, 2007, 74(4): 412-419

[9] Ty L V, Hanh N V, Uoc N T, Duc N H, Thanh N T, Huu Q X, Nguyen B X. Preliminary results of cell cryobanking and embryo production of black bear (Ursus thibetanus) by interspecies somatic cell nuclear transfer. Theriogenology, 2003, 59: 290

[10] White K L, Bunch T D, Mitalipov S, Reed W A. Establishment of pregnancy after the transfer of nuclear transfer embryos produced from the fusion of argali (Ovis ammon) nuclei into domestic sheep (Ovis aries) enucleated oocytes. Cloning, 1999, 1(1): 47-54

[11] Sansinena M J, Hylan D, Hebert K, Denniston R S, Godke R A. Banteng (Bos javanicus) embryos and pregnancies produced by interspecies nuclear transfer. Theriogenology, 2005, 63(4): 1081-1091

[12] Ikumi S, Sawai K, Takeuchi Y, Iwayama H, Ishikawa H, Ohsumi S, Fukui Y. Interspecies somatic cell nuclear transfer for in vitro production of Antarctic minke whale (Balaenoptera bonaerensis) embryos. Cloning and Stem Cells, 2004, 6(3): 284-293

[13] Williams J B, Shin T, Liu L, Flores-Foxworth G, Romano J, Blue-McClendon A, Kraemer D, Westhusin M E. Cloning of exotic/endangered species: desert bighorn sheep. Methods in Molecular Biology, 2006, 348: 169-181

[14] Li Y, Dai Y, Du W, Zhao C, Wang L, Wang H, Liu Y, Li R, Li N. In vitro development of yak (Bos grunniens) embryos generated by interspecies nuclear transfer. Animal Reproduction Science, 2007, 101(1-2): 45-59

[15] Li Y, Dai Y, Du W, Zhao C, Wang H, Wang L, Li R, Liu Y, Wan R, Li N. Cloned endangered species takin (Budorcas taxicolor) by inter-species nuclear transfer and comparison of the blastocyst development with yak (Bos grunniens) and bovine. Molecular Reproduction and Development, 2006, 73(2): 189-195

[16] Song J, Hua S, Song K, Zhang Y. Culture, characteristics and chromosome complement of Siberian tiger fibroblasts for nuclear transfer. In Vitro Cellular& Developmental Biology-Animal, 2007, 43(7): 203-209

[17] Damiani P, Wirtu G, Miller F. Development of gianteland (Taurotragus oryx) and bovine (Bos taurus) oocytes. Theriogenology, 2007, 59: 390

[18] Tao Y, Liu J, Zhang Y, Zhang M, Fang J, Han W, Zhang Z, Liu Y, Ding J, Zhang X. Fibroblast cell line establishment, cryopreservation and interspecies embryos reconstruction in red panda (Ailurus fulgens). Zygote, 2009, 17(2): 117-124

[19] Bhuiyan M M, Suzuki Y, Watanabe H, Lee E, Hirayama H, Matsuoka K, Fujise Y, Ishikawa H, Ohsumi S, Fukui Y. Production of sei whale (Balaenoptera borealis) cloned embryos by inter- and intra-species somatic cell nuclear transfer. Journal of Reproduction and Development, 2010, 56(1): 131-139

[20] Vogel G. Cloned gaur a short-lived success. Science, 2001, 291(5503): 409

[21] Yang C Y, Li R C, Pang C Y, Yang B Z, Qin G S, Chen M T, Zhang X F, Huang F X, Zheng H Y, Huang Y J, Liang X W. Study on the inter-subspecies nuclear transfer of river buffalo somatic cell nuclei into swamp buffalo oocyte cytoplasm. Animal Reproduction Science, 2010, 121(1-2): 78-83

[22] Liu K, Wu X, Wei Z Y, Bai C L, Su G H, Li G P. Comparative of the cytogenetic characteristics of Pantholops hodgsonii and other three species in Bovidae. Acta Theriologica Sinica, 2014, 34(1): 71-79

[23] Xue L, Cheng L, Su G, Kang F, Wu X, Bai C, Zhang L, Li G P. Nuclear transfer procedures in the ovine can induce early embryo fragmentation and compromise cloned embryo development. Animal Reproduction Science, 2011, 126(3-4): 179-186

[24] Duan B, Cheng L, Gao Y, Yin F X, Su G H, Shen Q Y, Liu K, Hu X, Liu X, Li G P. Silencing of fat-1 transgene expression in sheep may result from hypermethylation of its driven cytomegalovirus (CMV) promoter. Theriogenology, 2012, 78(4): 793-802

[25] Li G P, Bunch T D, White K L, Rickords L, Liu Y, Sessions B R. Denuding and centrifugation of maturing bovine oocytes alters oocyte spindle integrity and the ability of cytoplasm to support parthenogenetic and nuclear transfer embryo development. Molecular Reproduction and Development, 2006, 73(4): 446-451

[26] Bai C, Liu H, Liu Y, Wu X, Cheng L, Bou S, Li G P. Diploid oocyte formation and tetraploid embryo development induced by cytochalasin B in bovine. Cellular Reprogramming, 2011, 13(1): 37-45

[27] Frankfurt O S, Krishan A. Identification of apoptotic cells by formamide-induced DNA denaturation in condensed chromatin. Journal of Histochemistry and Cytochemistry, 2001, 49(3): 369-378

[28] Zhao Z J, Ouyang Y C, Nan C L, Lei Z L, Song X F, Sun Q Y, Chen D Y. Rabbit oocyte cytoplasm supports development of nuclear transfer embryos derived from the somatic cells of the camel and Tibetan antelope. Journal of Reproduction and Development, 2006, 52(3): 449-459

[29] Dominko T, Mitalipova M, Haley B, Beyhan Z, Memili E, McKusick B, First N L. Bovine oocyte cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species. Biology of Reproduction, 1999, 60(6): 1496-1502

[30] Chen D Y, Wen D C, Zhang Y P, Sun Q Y, Han Z M, Liu Z H, Shi P, Li J S, Xiangyu J G, Lian L, Kou Z H, Wu Y Q, Chen Y C, Wang P Y, Zhang H M. Interspecies implantation and mitochondria fate of panda-rabbit cloned embryos. Biology of Reproduction, 2002, 67(2): 637-642

[31] Yang X, Smith S L, Tian X C, Lewin H A, Renard J P, Wakayama T. Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning. Nature Genetics, 2007, 39(3): 295-302

[32] Beebe L F, McIlfatrick S J, Nottle M B. Cytochalasin B and trichostatin a treatment postactivation improves in vitro development of porcine somatic cell nuclear transfer embryos. Cloning and Stem Cells, 2009, 11(4): 477-482

[33] Bui H T, Wakayama S, Kishigami S, Park K K, Kim J H, Thuan N V, Wakayama T. Effect of trichostatin A on chromatin remodeling, histone modifications, DNA replication, and transcriptional activity in cloned mouse embryos. Biology of Reproduction, 2010, 83(3): 454-463

[34] Wittayarat M, Namula Z, Luu V, Do L T, Sato Y, Taniguchi M, Otoi T. Effect of trichostatin A on in vitro embryo development of interspecies nuclear transfer embryos reconstructed from cat donor nuclei and bovine cytoplasm. Reproduction, Fertility, and Development, 2012, 25(1): 161

[35] Hofmann J F, Sykora M, Redemann N, Beug H. G1-Cdk activity is required for both proliferation and viability of cytokine-dependent myeloid and erythroid cells. Oncogene, 2001, 20(31): 4198-4208

[36] Van Blerkom J. Mitochondria as regulatory forces in oocytes, preimplantation embryos and stem cells. Reproductive Biomedicine Online, 2008, 16(4): 553-569

[37] Wen D C, Bi C M, Xu Y, Yang C X, Zhu Z Y, Sun Q Y, Chen D Y. Hybrid embryos produced by transferring panda or cat somatic nuclei into rabbit MII oocytes can develop to blastocyst in vitro. Journal of Experimental Zoology Part A: Comparative Experimental Biology, 2005, 303A(8): 689-697

[38] Song J, Hua S, Song K, Zhang Y. Culture, characteristics and chromosome complement of Siberian tiger fibroblasts for nuclear transfer. In Vitro Cellular & Developmental Biology-Animal, 2007, 43(7): 203-209

Acknowledgements

We would like to thank Dr. Thomas D. Bunch of Animal, Dairy and Veterinary Sciences, Utah State University for his reading and editing this manuscript, thank Zhenjiang Zhang and Junlong Li who provide the domestic animals and animal facilities and thank Li Zhang, Liguo Zhang, Zehua Jiao, Shuhua Dong, Feng Kang, Biao Duan for their technical help. This work was supported by the National Basic Research Program of China (Grant No. 2012CB22306) and the National Natural Science Foundation of China (Grant No. 31372289).

Compliance with ethics guidelines

All applicable institutional and national guidelines for the care and use of animals were followed.

RIGHTS & PERMISSIONS

Higher Education Press and Springer-Verlag Berlin Heidelberg