Chromatin configuration in the germinal vesicle (GV) undergoes dynamic changes during oocyte growth, yet little is known about the mechanisms regulating chromatin remodeling in mouse oocytes. The hypothesis that companion granulosa cells play a role in modulating chromatin configuration and subsequent transcriptional activity in the oocyte genome was tested. Analysis of transcriptional activity, as determined by Br-UTP incorporation, revealed a similar percentage of transcriptionally active and inactive oocytes present in the large antral follicles of mature females. However, gonadotropin stimulation of follicular development induced an increase in the proportion of transcriptionally inactive oocytes. Interestingly, a similar proportion of stage-matched, oocyte-granulosa cell complexes grown in vitro without gonadotropin stimulation displayed chromatin redistribution around the nucleolus and no transcriptional activity. In contrast, when cultured in the absence of companion granulosa cells, transcriptional activity remained unabated in the majority of denuded GV stage oocytes. Extended prophase arrest in fully grown transcriptionally inactive oocyte-granulosa cell complexes had no effect on the progression of meiosis after in vitro maturation. However, it reduced the competence to complete preimplantation embryo development. These results indicate that chromatin redistribution around the nucleolus is associated with transcriptional repression in the GV of both fully grown in vivo-derived oocytes and cultured oocyte-granulosa cell complexes. Moreover, the results presented here suggest that some aspects of intraovarian control mechanisms were abrogated during culture of oocyte-granulosa cell complexes, resulting in a higher proportion of oocytes with "mature" chromatin. Most importantly, companion granulosa cells played an active role in modulating the transcriptional activity of the oocyte genome.
Copyright 2001 Academic Press.