Gene expression differences in oocytes derived from adult and prepubertal Japanese Black cattle during in vitro...

The present study was carried out to compare the gene expression profiles in oocytes derived from adult and prepubertal Japanese Black cattle during in vitro maturation (IVM) using microarray gene chips (Bovine genome array containing 24,072 probe sets representing over 23,000 transcripts). Microarray experiments were conducted using total RNA isolated from immature [germinal vesicle (GV)] and in vitro matured [metaphase II, (MII)] oocytes derived from adult and prepubertal animals. A total of 333 (1.4%) and 549 (2.3%) genes were differentially expressed between prepubertal vs adult bovine GV and MII stages oocytes, respectively. Of these, 176 and 312 genes were up-regulated, while 157 and 237 were down-regulated in prepubertal when compared with adult GV and MII oocytes, respectively. It was also observed that 695 (2.9%) and 553 (2.3%) genes were differentially expressed between GV vs MII stage oocytes in the adult and prepubertal groups, respectively. Gene ontological classification of the differentially expressed genes revealed that up-regulated genes in adult oocytes were involved in signal transduction, transcriptional control and transport. Quantitative reverse transcription-PCR validated the expression profile of some selected transcripts and confirmed differences in the expression levels of transcripts between adult vs prepubertal groups in both GV and MII stages oocytes as identified by microarray data analysis. This study indicated for the first time that significant number of genes were differentially expressed (>2-fold, p < 0.01) between oocytes derived from adult and those from prepubertal Japanese Black cattle, and this difference increased during IVM.