Ultrastructural organization, sites of transcription and distribution of fibrillar centres in the nucleolus of the mouse oocyte

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Abstract
The emergence of newly formed nucleoli and their development have been studied in mouse oocytes from pachytene to diplotene stages. At mid-pachytene, the nucleolus first appears as a fibrillar centre surrounded by a layer of electron-dense fibrils and penetrated by chromatin fibres emanating from the secondary constriction region of the nucleolar bivalent. Since this bivalent contains 2 paired nucleolar organizers, 2 nucleoli are formed in a symmetrical fashion. At advanced pachytene, the nucleoli are extended by strands of fibrillar component which become fibrillogranular distally. The 2 nucleoli fuse together at late pachytene. At diplotene, the nucleolus becomes large and reticulated. The development of the nucleolonema coincides with the appearance of numerous secondary fibrillar centres. Three-dimensional reconstruction of the reticulated nucleolus shows that the number of fibrillar centres largely exceeds that of nucleolar organizers. Radioautography after [‘HJuridine incorporation demonstrates that during the first step of nucleologenesis the labelling is limited to the layer of electron-dense fibrils surrounding the fibrillar centre. Study of the time course of tritiated uridine incorporation from pachytene to diplotene shows that the labelling extends with the extending strands of fibrillar component. In the fully developed nucleolus, all fibrillar strands are labelled and contain, therefore, actively transcribed rDNA. These observations suggest that the rDNA, which is initially compacted in the primary fibrillar centre at the onset of nucleogenesis, progressively unravels and becomes distributed throughout the fibrillar parts of the nucleolonema. The lack of labelling of the secondary fibrillar centres suggests that they are zones of inactivity of the ribosomal genes where the rDNA remains locally compacted. A model of the ultrastructural organization of the nucleolus is proposed based on our observations.