Oocyte morphology and transcriptional silencing in relation to chromatin remodeling during the final phases of bovine oocyte growth.

The complexity of the events which orchestrate mammalian oocyte growth and ultimate acquisition of developmental competence is still unclear and under continuous investigation. Starting from the observation that germinal vesicle (GV) bovine oocytes exhibit different patterns of chromatin configuration (from GV0 to GV3), the present study aimed at analyzing the morphology of the nuclear and the cytoplasmic compartments and to determine the total transcriptional activity of immature oocytes sorted on the basis of their chromatin configuration. The oocyte morphology was analyzed by light microscopy (LM) on semi-thin sections and transmission electron microscopy, and the global transcriptional activity was analyzed by H(3)-Uridine incorporation and subsequent autoradiography on semi-thin sections. LM confirmed the increase in chromatin condensation from GV0 to GV3. Ultrastructurally, the nucleolus was fibrillo granular at GV0 while the other stages displayed an electron-dense fibrillar sphere with the remnant of a fibrillar center on the surface. Organelles were dispersed in the cytoplasm at GV0 while at GV1 and GV2 most of them were homogenously distributed in the oocyte cortex. At GV3 most organelles were found in clusters in the oocyte cortex. Typical features of completion of the oocyte growth phase, like undulation of the nuclear envelope and reduction of the size of Golgi complex were found at GV2 and GV3. Moreover, GV3 oocytes presented cortical granules that displayed varying degrees of degeneration. Autoradiographic labeling showed that oocytes with GV0 configuration exhibited high level of RNA synthesis, GV1 and GV2 stages showed a remarkable decrease of transcription, and the acquisition of GV3 configuration was associated with global repression of transcriptional activity. Our findings suggest a temporal relationship between the chromatin remodeling process and the main morpho-functional events that characterize the final growth phase in bovine oocyte.