Bovine ooplasm partially remodels primate somatic nuclei following somatic cell nuclear transfer.
Interspecies somatic cell nuclear transfer (iSCNT) has the potential to become a useful tool to address basic questions about the nucleus-cytoplasm interactions between species. It has also been proposed as an alternative for the preservation of endangered species and to derive autologous embryonic stem cells. Using chimpanzee/ bovine iSCNT as our experimental model we studied the early epigenetic events that take place soon after cell fusion until embryonic genome activation (EGA). Our analysis suggested partial EGA in iSCNT embryos at the eight-cell stage, as indicated by Br-UTP incorporation and expression of chimpanzee embryonic genes. Oct4, Stella, Crabp1, CCNE2, CXCL6, PTGER4, H2AFZ, c-MYC, KLF4, and GAPDH transcripts were expressed, while Nanog, Glut1, DSC2, USF2, Adrbk1, and Lin28 failed to be activated. Although development of iSCNT embryos did not progress beyond the 8- to 16-cell stage, chromatin remodeling events, monitored by H3K27 methylation, H4K5 acetylation, and global DNA methylation, were similar in both intra- and interspecies SCNT embryos. However, bisulfite sequencing indicated incomplete demethylation of Oct4 and Nanog promoters in eight-cell iSCNT embryos. ATP production levels were significantly higher in bovine SCNT embryos than in iSCNT embryos, TUNEL assays did not reveal any difference in the apoptotic status of the nuclei from both types of embryos. Collectively, our results suggest that bovine ooplasm can partially remodel chimpanzee somatic nuclei, and provides insight into some of the current barriers iSCNT must overcome if further embryonic development is to be expected.
Wang
Kai
K
Michigan State University, East Lansing, 48824, USA.
Beyhan
Zeki
Z
Rodriguez
Ramon M
RM
Ross
Pablo J
PJ
Iager
Amy E
AE
Kaiser
German G
GG
Chen
Ying
Y
Cibelli
Jose B
JB
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D065150:Cellular Reprogramming; D019976:Cloning, Organism; D003593:Cytoplasm / Q000378:metabolism*; D019175:DNA Methylation; D044127:Epigenesis, Genetic / Q000235:genetics / Q000502:physiology; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D006657:Histones / Q000378:metabolism; D007621:Karyotyping; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D002679:Pan troglodytes
D006657:Histones