Inter-Specific Nucleartransfer Of Tibetan Antelope (Pantholops Hodgsoni)

Tibetan antelope is the unique wildlife in China, which mainly distributes in Tibetan area including Qinghai, Tibet and Xinjiang. Their populations decrease significantly due to the changes of environment and over hunting. It is urgent to protect this valuably species through every ways including modern biotechnology.In the present study, the fibroblast cells derived from a Tibetan antelope were transferred into enucleated bovine, ovine, and goat oocytes. The reconstructed inter-specific cloned embryos were cultured in vitro, or in the oviducts of sheep, goat, and rabbit, respectively. The results showed that the cleavage rates of the cloned embryos of Tibetan antelope-cattle and Tibetan antelope-sheep were 56% and 64%, respectively, which were significantly lower than that from bovine-bovine (75%) and sheep-sheep (81%). In the in vitro condition, around 0.47% of the Tibetan antelope-bovine reconstructed embryos develop to blastocysts. When goat, sheep and rabbit were used as intermediate animals, transfer of the inter-specific cloned embryos of Tibetan antelope-bovine,-goat, and-sheep to the oviducts of the recipient animals resulted in the similar embryo development to the in vitro culture. After transfer 429 Tibetan antelope- sheep cloned embryos to sheep oviducts, only 0.7% of the embryos developed to morula stage. Transfer of 168 Tibetan antelope-cattle cloned embryos to goat oviducts,1.6% developed to blastocysts. In order to improve the cloned embryo development, trichostatin A (TSA), an inhibitor of deacetylase, was used to treat the donor cells and bovine oocytes, the resultant cloned embryos were then transferred to the oviducts of sheep and goats. There are 10.5% embryos develop to the morula, but none of the embryos developed to blastocyst; while there were no Tibetan antelope-sheep cloned embryos developed to 8-cell stage. Roscovitine and TSA were further used to treat the bovine oocytes, respectively, and then the cloned Tibetan antelope-bovine embryos were transferred to rabbit oviducts. The roscovitine and TSA treated oocytes resulted in 2.3% and 14.3% embryos developed to blastocyst stage, respectively. In conclusion, Tibetan antelope-cattle reconstructed embryos could develop to blastocyst in vitro or in vivo, while goat and sheep oocytes did not support the cloned embryos developed to blastocyst stage.