J. Nurkowska

Abstract: We have shown that the extrachromosomal DNA body present in the oocyte nucleus of Creophilus maxillosus contains amplified copies of ribosomal DNA and that multiple nucleoli associated with extrachromosomal DNA body contain ribosomal RNA. In addition, we assessed the level of rDNA amplification in Creophilus oocytes. The amount of DNA in single extra DNA body corresponds to 86% of total nuclear DNA content or 617% of chromosomal DNA content. In Creophilus, there are two phases of transcriptional activity of extrachromosomal rDNA, first in oogonia, and second in growing oocytes, separated by a short period of transcriptional quiescence in the ovary of newly hatched imago. We have described the organization and activity of extra DNA body in RNA synthesis and production of multiple nucleoli in previtellogenic stages of oocyte growth in imaginal ovariole of Creophilus.

Abstract: Nucleoli disassemble in prophase of the metazoan mitotic cycle, and they begin their reassembly (nucleologenesis) in late anaphase?early telophase. Nucleolar disassembly and reassembly were obvious to the early cytologists of the eighteenth and nineteenth centuries, and although this has lead to a plethora of literature describing these events, our understanding of the molecular mechanisms regulating nucleolar assembly and disassembly has expanded immensely just within the last 10-15 years. We briefly survey the findings of nineteenth-century cytologists on nucleolar assembly and disassembly, followed by the work of Heitz and McClintock on nucleolar organizers. A primer review of nucleolar structure and functions precedes detailed descriptions of modern molecular and microscopic studies of nucleolar assembly and disassembly. Nucleologenesis is concurrent with the reinitiation of rDNA transcription in telophase. The perichromosomal sheath, prenucleolar bodies, and nucleolar-derived foci serve as repositories for nucleolar processing components used in the previous interphase. Disassembly of the perichromosomal sheath along with the dynamic movements and compositional changes of the prenucleolar bodies and nucleolus-derived foci coincide with reactivation of rDNA synthesis within the chromosomal nucleolar organizers during telophase. Nucleologenesis is considered in various model organisms to provide breadth to our understanding. Nucleolar disassembly occurs at the onset of mitosis primarily as a result of the mitosis-specific phosphorylation of Pol I transcription factors and processing components. Although we have learned much regarding nucleolar assembly and disassembly, many questions still remain, and these questions are as vibrant for us today as early questions were for nineteenth- and early twentieth-century cytologists.

Abstract: The ovaries of the studied earwig species (Forficula auricularia, Chelidurella acanthopygia, Doru lineare and Opisthocosmia silvestris) are meroistic-polytrophic and composed of numerous short ovarioles that consist of a terminal filament, germarium and vitellarium. The germaria of adult females comprise meiotic (pachytene) and postmeiotic (differentiating) germ cell clusters, as well as small prefollicular cells. All germ cell clusters consist of two cells that are connected by a single intercellular bridge. In the vitellarium there are usually 2 ovarian follicles only. The individual follicle consists of a transcriptionally dormant oocyte and a single, polyploid nurse cell and is surrounded by a layer of somatic follicular cells (FCs). During previtellogenesis the nurse cell enlarges and becomes highly transcriptionally active. Concurrently its nucleus attains a characteristic, irregular shape. In the nurse cell nucleus of one studied species, F. auricularia, in addition to chromatin aggregations and RNA- and Ag-NOR-positive nucleoli, a single compact DNA-positive body is present. During advanced vitellogenesis the molecules synthesized in the nurse cells (RNAs, proteins, as well as nurse cell organelles) are transferred to the ooplasm via the intercellular bridge. During this transfer the nurse cell nucleus is retained in the cell centre and does not occlude the intercellular bridge. The results of our studies indicate that such position of the nurse cell nucleus is maintained solely by its extended shape. In other words, the rigid extensions keep the nucleus in the cell centre while the cytoplasm flows, in between these extensions, towards the intercellular bridge connecting the nurse cell with the oocyte.

Abstract: Oogenesis in the glossiphoniid leech Glossiphonia heteroclita (Hirudinea, Rhynchobdellida) is nutrimental, i.e., the growing oocyte is supported by specialized germline cells, the nurse cells. The main function of the nurse cells is to provide oocytes with cell organelles and RNAs (mainly rRNA). However, in studied leech species, irrespective of the nutrimental mode of oogenesis, the germinal vesicle (GV = oocyte nucleus) seems to be very active in rRNA production. As shown in the present study, during early previtellogenesis in the GV the meiotic chromosomes and prominent primary nucleoli occur. In late previtellogenesis the chromosomes condense and occupy a limited space of nucleoplasm in close vicinity to primary nucleolus, forming a karyosome. At the onset of vitellogenesis several prominent extrachromosomal DNA bodies appear in close association with the karyosome. At the same time, the primary nucleolus is no longer visible in the GV. As vitellogenesis proceeds the extrachromosomal DNA bodies undergo fragmentation and numerous spherical, RNA- and AgNOR-positive inclusions occur in the nucleoplasm. They are regarded as multiple nucleoli. Finally, in late oogenesis numerous accessory nuclei are formed in close proximity to the nuclear envelope. They usually contain one dense body, morphologically similar to multiple nucleoli. The amplification of rDNA genes, the occurrence of extrachromosomal DNA bodies, as well as the presence of multiple nucleoli and accessory nuclei are described for the first time in the phylum Annelida. J. Morphol. © 2005 Wiley-Liss, Inc.

Abstract: Preliminary histological analysis showed that the ovaries in Nannochorista neotropica are composed of numerous ovarioles of panoistic type. In the ovaries of adult females each ovariole consists of a terminal filament, vitellarium and pedicle while a germarium is absent. Morphological observations supported by simple histochemical tests revealed the presence of multiple nucleoli in the oocyte nucleus. The appearance of multiple nucleoli suggests that the extrachromosomal amplification of ribosomal DNA may take place in the oocytes of Nannochorista. The results show that in the structure of the ovariole and the course of oogenesis nannochoristids share noteworthy derived features with boreids, but differ significantly from all other mecopterans. These findings indicate the possible monophyly of an assemblage comprising the Nannomecoptera, Neomecoptera and Siphonaptera.

Abstract: The ovaries of the snow scorpionfly, Boreus hyemalis (Mecoptera : Boreidae) are panoistic and comprise 7–8 ovarioles. Each ovariole consists of a terminal filament, elongated vitellarium, and ovariole stalk (=pedicel) only ; in adult specimens, functional germaria are absent. Five consecutive stages of oogenesis i.e., early, mid- and late previtellogenesis, vitellogenesis, and choriogenesis have been distinguished in imagines. Oocyte nuclei (=germinal vesicles) of previtellogenic oocytes contain numerous polymorphic multiple nucleoli (or nucleolar masses), endobodies, and chromatin aggregations. Next to the nuclear envelope, large accumulations of nuage material are localized. The ooplasm of late previtellogenic oocytes is differentiated into transparent (perinuclear) and opaque (peripheral) regions. Ultrastructural investigations have revealed that within the latter, abundant ribosomes as well as mitochondria, elements of endoplasmic reticulum, Golgi complexes, annulate lamellae, symbiotic bacteroids, lipid droplets and distinctive accumulations of membrane-free clathrin-like cages are present. Early- and mid previtellogenic oocytes are invested with flat somatic cells that gradually transform into a follicular epithelium. In the vicinity of 3-cell junctions, neighbouring follicular cells are joined by narrow intercellular bridges. During late previtellogenesis, numerous microvilli develop on the oocyte surface. They interdigitate with morphologically similar but less frequent microvilli of the follicular cells. Concurrently, first endocytotic vesicles appear in the cortical ooplasm. In the context of presented results, the phylogenetic relationships between mecopterans (boreids) and fleas are discussed.