Differential gene expression in bovine elongated (Day 17) embryos produced by somatic cell nucleus transfer and in vitro fertilization.
Somatic cloning in cattle is associated with impaired embryo development, caused by inappropriate epigenetic reprogramming during embryogenesis; however, there is a paucity of data regarding gene expression at the critical elongation and peri-implantation stages. The objective of the present study was to identify genes differentially expressed in bovine cloned embryos at Day 17 of development (Day 0=day of nucleus transfer or IVF). Day 7 blastocysts (Hand Made Cloned or IVP) were transferred to recipient cattle and collected at Day 17. The efficiency of recovery of elongated embryos was similar, however cloned embryos elongated less than IVP embryos (91.8+/-45.8 vs. 174+/-50mm) and fewer had embryonic discs (63 vs. 83%). Qualitative and quantitative PCR detected expression of OCT4, NANOG, IFNtau, EOMES, FGF4, SOX2, and CDX2 in all IVP embryos. In most cloned embryos, NANOG and FGF4 were absent (verified by qPCR); NANOG, EOMES, and FGF4 were underexpressed, whereas IFNtau was overexpressed in cloned embryos. Based on qPCRs, other genes, i.e., SPARC, SNRB1, and CBPP22, were down-regulated in cloned embryos, whereas HSP70 and TDKP1 were overexpressed. In bovine microarrays, 47 genes (3.6%) were deregulated in cloned embryos, including several involved in trophoblast growth and differentiation. In conclusion, we inferred that these data were indicative of incomplete epigenetic reprogramming after cloning; this could lead to aberrant gene expression and subsequently early pregnancy loss. There was an apparent association between incomplete morphological elongation and aberrant reprogramming of a subset of genes critical for early embryonic development.
Rodríguez-Alvarez
Lleretny
L
Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Campus Chillán, Avenida Vicente Méndez 595, Chillán, Chile.
Sharbati
Jutta
J
Sharbati
Soroush
S
Cox
José Francisco
JF
Einspanier
Ralf
R
Castro
Fidel Ovidio
FO
D000818:Animals; D002417:Cattle / Q000196:embryology*; D019976:Cloning, Organism / Q000662:veterinary*; D046149:Embryo Culture Techniques / Q000662:veterinary; D004624:Embryo Transfer / Q000662:veterinary; D004622:Embryo, Mammalian / Q000378:metabolism*; D047108:Embryonic Development / Q000235:genetics; D044127:Epigenesis, Genetic / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005865:Gestational Age; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011247:Pregnancy