Analysis of Differential Messenger RNA Expression Between Bovine Blastocysts Produced in Different Culture Systems: Implications for Blastocyst Quality

摘要: Using reverse transcriptase-amplified fragment length polymorphism (RT-AFLP) analysis of differential mRNA expression and semiquantitative transcriptase-polymerase chain reaction, we compared in bovine blastocysts from 4 sources, known to differ quality terms their ability withstand cryopreservation: 1) vitro culture synthetic oviduct fluid vitro-matured (IVM)/in fertilized (IVF) zygotes; 2) TCM-199 supplemented with granulosa cells (coculture) IVM/IVF 3) vivo the ewe or 4) superovulation, artificial insemination, nonsurgical embryo recovery. Total was isolated pools transcription performed. Triplicate reactions each sample were displayed, only consistent banding variations recorded. AFLP-differential display assay, found that cDNA patterns are highly conserved between groups studied; however, there a difference 7% bands either missing expressed across groups. Fifty reamplified, sequence comparison search revealed similarity 14 fragments ribosomal mitochondrial genes, 16 matched described cDNA, 20 corresponded unknown sequences may represent novel genes. The study 7 differentially mRNAs be involved developmental process suggests roles for apoptosis, oxidative stress, gap junctions, differentiation determination quality. aberrant detected vitro-produced embryos those produced explain reduced viability after cryopreservation.