Development of bovine 13;ovine interspecies cloned embryos and mitochondria segregation in blastomeres during preimplantation
- 作者：Song Hua ; Yong Zhang ; Kai Song ; Jimei Song ; Zhipeng Zhang ; Lin Zhang ; Chi Zhang ; Junwei Cao ; Libing Ma
- 关键词：Bovine ; Ovine ; Nuclear transfer ; Mitochondrion ; Real-time PCR
- 刊名：Animal Reproduction Science
- 出版年：2008
- 期刊代码：8_03784320
- 类别：abs
- 出版时间：May 2008
- 卷：105
- 期：3-4
- 页码：245-257
- 文件大小：208 K
摘要
The objective of the study was to investigate interspecies somatic cell nuclear transfer (iSCNT) embryonic potential and mitochondrial DNA (mtDNA) segregation during preimplantation development. We generated bovine 13;ovine reconstructed embryos via iSCNT using bovine oocytes as recipient cytoplasm and ovine fetal fibroblast as donor cells. Chromosome composition, the total cell number of blastocyst and embryonic morphology were analyzed. In addition, mtDNA copy numbers both from donor cell and recipient cytoplasm were assessed by real-time PCR in individual blastocysts and blastomeres from 1- to 16-cell stage embryos. The results indicated the following: (1) cell nuclei of ovine fetal fibroblasts can dedifferentiate in enucleated bovine ooplasm, and the reconstructed embryos can develop to blastocysts. (2) 66%of iSCNT embryos had the same number of chromosome as that of donor cell, and the total cell number of iSCNT blastocysts was comparable to that of sheep parthenogenetic blastocysts. (3) RT-PCR analysis in individual blastomeres revealed that the ratio of donor cell mtDNA: recipient cytoplasm mtDNA remained constant (1%) from the one- to eight-cell stage. However, the ratio decreased from 0.6%at the 16-cell stage to 0.1%at the blastocyst stage. (4) Both donor cell- and recipient cytoplasm-derived mitochondria distributed unequally in blastomeres with progression of cell mitotic division. Considerable unequal mitochondrial segregation occurred between blastomeres from the same iSCNT embryos.