CUSTOM MICROARRAY ANALYSIS OF GENE EXPRESSION OF OVINE OOCYTES SUBJECTED TO VITRIFICATION

Aim of this work was to evaluate the effects of vitrification procedures. on the transcriptome of ovine oocytes using a customized ovine cDNA microarray.. The analysis was performed on vitrified-warmed in vitro matured oocytes (VTR) and on. control oocytes (CTR) subjected only to in vitro maturation. Vitrification was. performed with the minimum essential volume method using cryotop as device. (Kuwayama et al., 2005).. We generated a medium density microarray based on Illumina DASL technology,. comprising 1536 independent assays addressing 1039 transcripts relevant to several cell. functions.. VTR and IVM oocytes were subjected to mRNA isolation and subsequently hybridized. to the customized microarray. Comparison of mRNA content between VTR and CTR. oocytes revealed 29 differentially expressed genes, mainly down-regulated (25. transcripts) after vitrification (P<0.01). Microarray analysis results were validated by. quantitative reverse transcription Real Time PCR of six selected transcripts.. This study highlights alterations in the transcriptome of ovine oocytes as a consequence. of vitrification procedures. The identification of the molecular mechanisms involved in. the oocyte reaction to cryopreservation is an important step for the improvement of the. experimental procedures. ""