Nuclear lamin antigen and messenger RNA expression in bovine in vitro produced and nuclear transfer embryos.
The nuclear lamina is a complex meshwork of nuclear lamin filaments that lies on the interface of the nuclear envelope and chromatin and is important for cell maintenance, nucleoskeleton support, chromatin remodeling, and protein recruitment to the inner nucleolus. Protein and mRNA patterns for the major nuclear lamins were investigated in bovine in vitro fertilized (IVF) and nuclear transfer embryos. Expression of lamins A/C and B were examined in IVF bovine germinal vesicle (GV) oocytes, metaphase II oocytes, zygotes, 2-cell, 8-cell, 16-32-cell embryos, morulae, and blastocysts (n = 10). Lamin A/C was detected in 9/10 immature oocytes, 10/10 zygotes, 8/10 2-cell embryos, 4/10 morulae, 10/10 blastocysts but absent during the maternal embryonic transition. Lamin B was ubiquitously expressed during IVF preimplantation development but was only detected in 4/10 GV oocytes. Messenger RNA expression confirms that the major lamins, A/C and B1 are expressed throughout preimplantation development and transcribed by the embryo proper. Lamin A/C and B expression were observed (15 min, 30 min, 60 min, 120 min) following somatic cell nuclear transfer using adult fibroblasts and at the 2-cell, 8-cell, 16-32-cell, morula and blastocyst stage (n = 5). Altered expression levels and localization of nuclear lamins A/C and B was determined in nuclear transfer embryos during the first 2 hr post fusion, coincidental with only partial nuclear envelope breakdown as well as during the initial cleavage divisions, but was restored by the morula stage. This mechanical and molecular disruption of the nuclear lamina provides key evidence for incomplete nuclear remodeling and reprogramming following somatic cell nuclear transfer.
Hall
Vanessa J
VJ
Centre for Early Human Development, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia.
Cooney
Melissa A
MA
Shanahan
Prue
P
Tecirlioglu
R Tayfur
RT
Ruddock
Nancy T
NT
French
Andrew J
AJ
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D019976:Cloning, Organism / Q000379:methods; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D034882:Lamins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D009028:Morula / Q000166:cytology / Q000378:metabolism*; D009685:Nuclear Envelope / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D015053:Zygote / Q000166:cytology / Q000502:physiology*
D034882:Lamins; D012333:RNA, Messenger