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

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          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.

          Most cited references32

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          Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning.

          Therapeutic cloning, whereby somatic cell nuclear transfer (SCNT) is used to generate patient-specific embryonic stem cells (ESCs) from blastocysts cloned by nuclear transfer (ntESCs), holds great promise for the treatment of many human diseases. ntESCs have been derived in mice and cattle, but thus far there are no credible reports of human ntESCs. Here we review the recent literature on nuclear reprogramming by SCNT, including studies of gene expression, DNA methylation, chromatin remodeling, genomic imprinting and X chromosome inactivation. Reprogramming of genes expressed in the inner cell mass, from which ntESCs are derived, seems to be highly efficient. Defects in the extraembryonic lineage are probably the major cause of the low success rate of reproductive cloning but are not expected to affect the derivation of ntESCs. We remain optimistic that human therapeutic cloning is achievable and that the derivation of patient-specific ntESC lines will have great potential for regenerative medicine.
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            Effect of trichostatin A on chromatin remodeling, histone modifications, DNA replication, and transcriptional activity in cloned mouse embryos.

            Our group and others have found that the treatment of embryos with trichostatin A (TSA) after cloning by somatic cell nuclear transfer (SCNT) results in a significant improvement in efficiency. We believe that TSA treatment improves nuclear remodeling via histone modifications, which are important in the epigenetic regulation of gene silencing and expression. Some studies found that treatment of SCNT-generated embryos with TSA improved lysine acetylation of core histones in a manner similar to that seen in normally fertilized embryos. However, how histone methylation is modified in TSA-treated cloned embryos is not completely understood. In the present study, we found that TSA treatment caused an increase in chromosome decondensation and nuclear volume in SCNT-generated embryos similar to that in embryos produced by intracytoplasmic sperm injection. Histone acetylation increased in parallel with chromosome decondensation. This was associated with a more effective formation of DNA replication complexes in treated embryos. We also found a differential effect of TSA on the methylation of histone H3 at positions K4 and K9 in SCNT-generated embryos that could contribute to genomic reprogramming of the somatic cell nuclei. In addition, using 5-bromouridine 5'-triphosphate-labeled RNA, we showed that TSA enhanced the levels of newly synthesized RNA in 2-cell embryos. Interestingly, the amount of SCNT-generated embryos showing asymmetric expression of nascent RNA was reduced significantly in the TSA-treated group compared with the nontreated group at the 2-cell stage. We conclude that the incomplete and inaccurate genomic reprogramming of SCNT-generated embryos was improved by TSA treatment. This could enhance the reprogramming of somatic nuclei in terms of chromatin remodeling, histone modifications, DNA replication, and transcriptional activity.
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              Mitochondria as regulatory forces in oocytes, preimplantation embryos and stem cells.

              In addition to their role in generating energy (ATP), other possible regulatory functions for mitochondria in the mature oocyte, preimplantation stage embryo and differentiating embryonic stem cell are discussed. The question of the numerical size of a normal mitochondrial complement in the mature oocyte, which has been suggested to be a critical factor in the determination of oocyte and embryo developmental competence, is addressed in the context of mitochondrial DNA copy numbers and the apparent number of organelles detected in living mouse and human blastocysts with mitochondria-specific fluorescent probes. The existence of a spatially stable subplasmalemmal domain of high-polarized mitochondria in the mature oocyte and early embryo is proposed to form a microzone of differential activity that may locally influence the normality of fertilization. High-polarized mitochondria may also influence developmentally significant activities in the peri-implantation blastocyst and differentiating embryonic stem cell. The work discussed indicates that mitochondria may have multiple functions during early development, and specific areas of investigation required to confirm these possibilities are suggested.
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                Author and article information

                Contributors
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                Journal
                Front Agric Sci Eng
                FASE
                CN10-1204/S
                Frontiers of Agricultural Science and Engineering
                Higher Education Press (4 Huixin Dongjie, Chaoyang District, Beijing 100029, China )
                2095-7505
                2014
                : 1
                : 1
                : 28-36
                Affiliations
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Key Laboratory of Herbivore Reproductive Biotechnology and Breeding Ministry of Agriculture, Inner Mongolia University, Hohhot 010021, China
                Author notes
                xurg@cae.cn
                Article
                10.15302/J-FASE-2014012
                567d6390-c625-4b21-8e54-b642ed36b618
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 03 March 2014
                : 16 March 2014
                Categories
                RESEARCH ARTICLE

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

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