Aberrant DNA methylation in porcine in vitro-, parthenogenetic-, and somatic cell nuclear transfer-produced blastocysts.
Early embryonic development in the pig requires DNA methylation remodeling of the maternal and paternal genomes. Aberrant remodeling, which can be exasperated by in vitro technologies, is detrimental to development and can result in physiological and anatomic abnormalities in the developing fetus and offspring. Here, we developed and validated a microarray based approach to characterize on a global scale the CpG methylation profiles of porcine gametes and blastocyst stage embryos. The relative methylation in the gamete and blastocyst samples showed that 18.5% (921/4,992) of the DNA clones were found to be significantly different (P < 0.01) in at least one of the samples. Furthermore, for the different blastocyst groups, the methylation profile of the in vitro-produced blastocysts was less similar to the in vivo-produced blastocysts as compared to the parthenogenetic- and somatic cell nuclear transfer (SCNT)-produced blastocysts. The microarray results were validated by using bisulfite sequencing for 12 of the genomic regions in liver, sperm, and in vivo-produced blastocysts. These results suggest that a generalized change in global methylation is not responsible for the low developmental potential of blastocysts produced by using in vitro techniques. Instead, the appropriate methylation of a relatively small number of genomic regions in the early embryo may enable early development to occur.
Bonk
Aaron J
AJ
Division of Animal Science, University of Missouri, Columbia, Missouri 65211, USA.
Li
Rongfeng
R
Lai
Liangxue
L
Hao
Yanhong
Y
Liu
Zhonghua
Z
Samuel
Melissa
M
Fergason
Emily A
EA
Whitworth
Kristin M
KM
Murphy
Clifton N
CN
Antoniou
Eric
E
Prather
Randall S
RS
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002467:Cell Nucleus / Q000502:physiology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D019175:DNA Methylation; D017931:DNA Primers; D047108:Embryonic Development; D005260:Female; D007839:Functional Laterality; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D009865:Oocytes / Q000502:physiology; D010312:Parthenogenesis; D012741:Sexual Maturation; D013094:Spermatozoa / Q000502:physiology; D013552:Swine
D017931:DNA Primers; D004247:DNA