A Light- And Electron-Microscope Study of the Nucleolus During Growth of the Oocyte in the Prepubertal Mouse

1971 ◽  
Vol 9 (3) ◽  
pp. 637-663
Author(s):  
L. A. CHOUINARD
Keyword(s):  
Structural Organization ◽  
Functional Organization ◽  
Current Knowledge ◽  
Light And Electron Microscopy ◽  
Fold Increase ◽  
Dense Fibrillar Component ◽  
Parallel Increase ◽  
Nucleolar Material ◽  
Nucleolar Volume ◽  
Fibrillar Component

The ordered changes which occur in the structural organization of the nucleolus during growth of the mouse oocyte have been studied by both light and electron microscopy. All observations have been made on those oocytes whose growth is initiated on the day of birth and completed by postnatal day 14 in prepubertal animals of the ICR albino mouse strain. During that period the oocyte nucleolus undergoes an approximate 90-fold increase in volume. During the unilaminar follicle stage (from birth to postnatal day 4), the growing nucleolus exhibits an overall reticulated-type of structure consisting of: (1) a moderately electron-dense fibrillogranular component occupying most parts of the nucleolar framework; (2) an electron-transparent nucleoplasm-like component filling the numerous interstices of the nucleolar framework; (3) an electron-dense fibrillar component located in the peripheral portion of a number of small islands widely and uniformly scattered within the nucleolar framework, and (4) a slightly less-dense fibrillar component situated in the central portion of these same islands and referred to as fibrillar centres. Increase in nucleolar volume during that stage is brought about mainly through an increase in the overall dimensions of the fibrillogranular framework, accompanied by a parallel increase in the number and, to a certain extent, the size of its electron-transparent interstices. During the bilaminar follicle stage (postnatal day 5 through 8), the following structural and organizational changes take place more or less concomitantly within the still enlarging nucleolar mass: (1) the fibrillogranular framework becomes predominantly fibrillar in texture as a result of what appears to be an unravelling or unfolding of its constituent granules of ribosomal dimensions; (2) the nucleolar interstices decrease rapidly both in number and size because of the accumulation within their interior of a material the texture and density of which match that present in the nucleolar framework itself; and (3) a number of rounded electron-transparent spaces, the nucleolar vacuoles, make their appearance in the regions formerly occupied by some of the fibrillar islands and adjacent interstices. Increase in nucleolar volume during that stage is largely due to the appearance and subsequent enlargement of the nucleolar vacuoles in question. During the plurilaminar follicle stage (postnatal day 9 through 14), the following sequential events take place within the nucleolar mass: (1) a moderately electron-dense fibrillogranular material accumulates within the nucleolar vacuoles; (2) this fibrillogranular material, which eventually fills all vacuolar spaces, undergoes degranulation and a concomitant increase in density, eventually matching that of the rest of the nucleolar mass; (3) all remnants of the lightly stained nucleolar interstices disappear from view; and (4) the fully grown rounded nucleolus finally appears as a dense, compact mass, exclusively fibrillar in texture, and exhibiting no internal structural organization. An attempt is made to interpret these changes in the light of current knowledge concerning thearchitectural and functional organization of the mammalian nucleolus in general. The observations are consistent with the view that the nucleolus, during growth of the primary oocyte, is the site of massive synthesis and storage of nucleolar material.

A light- and electron-microscope study of the oocyte nucleus during development of the antral follicle in the prepubertal mouse

1975 ◽  
Vol 17 (3) ◽  
pp. 589-615
Author(s):  
L.A. Chouinard
Keyword(s):  
Meiotic Prophase ◽  
Functional Organization ◽  
Current Knowledge ◽  
Light And Electron Microscopy ◽  
Antral Follicle ◽  
Mouse Oocyte ◽  
Oocyte Nucleus ◽  
Follicle Development ◽  
Preovulatory Follicle ◽  
Functional Changes

The ordered changes which occur in the structural organization of the mouse oocyte nucelus during the preparatory, the maturative and the preovulatory stages of antral follicle development, have been studied under both light and electron microscopy. All observations have been made on those antral follicles whose development is initiated on postnatal day 14 and completed by postnatal day 28 in prepubertal animals of the ICR albino mouse strain. The formed entities that can be recognized within the oocyte nucleus during that period are the condensing bivalents, the heterochromatic knobs, the nucleolus and the extranucleolar bodies. At the onset of antral follicle development, the highly unravelled dictyate bivalents are seen to take on a lampbrush-type configuration. Subsequent condensation of these lampbursh bivalents appears to be a very gradual and lengthy process that extends over almost the entire period of antral follicle development. The shortening and thickening of the lampbrush bivalents are best interpreted as resulting from the withdrawal of their lateral loop-like projections into the chromosome axes and from the focal aggregation of these axes into compact chromatin masses. Electron-opaque granules, which appear within the oocyte nucleus during the preparatory and maturative follicle stages, are seen to be intimately associated with these condensing bivalents. A number of Feulgen-positive heterochromatic knobs make their appearance in contact with certain bivalents during the preparatory follicle stage. These knobs are not reincorporated as such into the condensing chromatin masses and undergo disintegration and dissolution during the preovulatory follicle stage. The size, shape and ultrastructural features of the nucleolus remain unchanged thoughout the period of antral follicle development. Breakdown and dissolution of the nucleolar mass is a swift process that takes place only in the fully mature preovulatory follicle and more or less concomitantly with the dismantling of the nuclear envelope. The extranucleolar bodies increase noticeably in size during the preparatory and the maturative follicle stages; they shrink in size and undergo dissolution during the preovulatory stage of antral follicle development. An attempt is made to interpret these morphological changes in the light of current knowledge concerning the architectural and functional organization of the oocyte nucleus in general during meiotic prophase. The relevant observational evidence would be consistent with the view that, during antral follicle deveopment, the mouse oocyte nucleus is not, as too often assumed, in a period of arrested evolution; its formed components undergo structural, maturational and functional changes which are of significance not only for the resumption of the first meiotic prophase but also for the early development of the embryo.

scholarly journals Inhibition of nucleolar reformation after microinjection of antibodies to RNA polymerase I into mitotic cells.

1987 ◽  
Vol 105 (4) ◽  
pp. 1483-1491 ◽  
Author(s):  
R Benavente ◽  
K M Rose ◽  
G Reimer ◽  
B Hügle-Dörr ◽  
U Scheer
Keyword(s):  
Rna Polymerase ◽  
Cultured Cells ◽  
Active Form ◽  
Nucleolar Organizer ◽  
Rna Polymerase I ◽  
Nucleolar Organizer Regions ◽  
Dense Fibrillar Component ◽  
Nucleolar Material ◽  
Fibrillar Component ◽  
Polymerase I

The formation of daughter nuclei and the reformation of nucleolar structures was studied after microinjection of antibodies to RNA polymerase I into dividing cultured cells (PtK2). The fate of several nucleolar proteins representing the three main structural subcomponents of the nucleolus was examined by immunofluorescence and electron microscopy. The results show that the RNA polymerase I antibodies do not interfere with normal mitotic progression or the early steps of nucleologenesis, i.e., the aggregation of nucleolar material into prenucleolar bodies. However, they inhibit the telophasic coalescence of the prenucleolar bodies into the chromosomal nucleolar organizer regions, thus preventing the formation of new nucleoli. These prenucleolar bodies show a fibrillar organization that also compositionally resembles the dense fibrillar component of interphase nucleoli. We conclude that during normal nucleologenesis the dense fibrillar component forms from preformed entities around nucleolar organizer regions, and that this association seems to be dependent on the presence of an active form of RNA polymerase I.

Nucleologenesis in Trypanosoma cruzi

2016 ◽  
Vol 22 (3) ◽  
pp. 621-629 ◽  
Author(s):  
Tomás Nepomuceno-Mejía ◽  
Reyna Lara-Martínez ◽  
Roberto Hernández ◽  
María de Lourdes Segura-Valdez ◽  
Luis F. Jiménez-García
Keyword(s):  
Cell Division ◽  
Trypanosoma Cruzi ◽  
Ethylenediaminetetraacetic Acid ◽  
Organizer Region ◽  
Nucleolar Organizer Region ◽  
Light And Electron Microscopy ◽  
Nucleolar Organizer ◽  
Nuclear Bodies ◽  
Nucleolar Organizer Regions ◽  
Nucleolar Material

AbstractNucleolar assembly is a cellular event that requires the synthesis and processing of ribosomal RNA, in addition to the participation of pre-nucleolar bodies (PNBs) at the end of mitosis. In mammals and plants, nucleolar biogenesis has been described in detail, but in unicellular eukaryotes it is a poorly understood process. In this study, we used light and electron microscopy cytochemical techniques to investigate the distribution of nucleolar components in the pathway of nucleolus rebuilding during closed cell division in epimastigotes of Trypanosoma cruzi, the etiologic agent of American trypanosomiasis. Silver impregnation specific for nucleolar organizer regions and an ethylenediaminetetraacetic acid regressive procedure to preferentially stain ribonucleoprotein revealed the conservation and dispersion of nucleolar material throughout the nucleoplasm during cell division. Furthermore, at the end of mitosis, the argyrophilic proteins were concentrated in the nucleolar organizer region. Unexpectedly, accumulation of nucleolar material in the form of PNBs was not visualized. We suggest that formation of the nucleolus in epimastigotes of T. cruzi occurs by a process that does not require the concentration of nucleolar material within intermediate nuclear bodies such as mammalian and plant PNBs.

A study on nucleolar DNA: isolation of DNA from fibrillar components and ultrastructural localization of different DNA probes

1993 ◽  
Vol 104 (4) ◽  
pp. 1199-1205 ◽  
Author(s):  
P. Hozak ◽  
C. Schofer ◽  
J. Sylvester ◽  
F. Wachtler
Keyword(s):  
Ribosomal Rna ◽  
Sertoli Cells ◽  
Ultrastructural Localization ◽  
Ribosomal Rna Genes ◽  
Border Region ◽  
Dense Fibrillar Component ◽  
Mitotic Chromosomes ◽  
Electron Microscopic ◽  
Fibrillar Component ◽  
Rna Genes

The nature and localization of DNA contained in the fibrillar centres and the dense fibrillar component (the fibrillar complex) in the nucleoli, was studied in human LEP cells, Sertoli cells, spermatogonia A and in mitotic chromosomes of stimulated lymphocytes. A novel procedure for isolating the intact fibrillar complex from LEP cells was used; the complex contains DNA that hybridizes to secondary constrictions of mitotic chromosomes and to 28 S rDNA sequences, on Southern blots. Electron microscopic DNA-DNA in situ hybridization was performed, with (a) a probe prepared from DNA extracted from the fibrillar complex of LEP cells, (b) a probe for human total genomic DNA, and (c) a probe for the transcribed part of human rDNA. On the basis of the results obtained we conclude that the ribosomal RNA genes in human Sertoli cells and spermatogonia A are predominantly associated with the dense fibrillar component, including the border region between fibrillar centres and the dense fibrillar component. The ribosomal RNA genes are the main, if not exclusive, DNA type present in the fibrillar complex in the studied cell types.

scholarly journals Mitochondrial Biogenesis and Remodeling during Adipogenesis and in Response to the Insulin Sensitizer Rosiglitazone

2003 ◽  
Vol 23 (3) ◽  
pp. 1085-1094 ◽  
Author(s):  
Leanne Wilson-Fritch ◽  
Alison Burkart ◽  
Gregory Bell ◽  
Karen Mendelson ◽  
John Leszyk ◽  
...  
Keyword(s):  
Mitochondrial Biogenesis ◽  
Gradient Centrifugation ◽  
Light And Electron Microscopy ◽  
Fold Increase ◽  
Mitochondrial Proteins ◽  
Whole Body ◽  
Brown Adipocyte ◽  
Insulin Sensitizer ◽  
Adipose Differentiation ◽  
Whole Body Metabolism

ABSTRACT White adipose tissue is an important endocrine organ involved in the control of whole-body metabolism, insulin sensitivity, and food intake. To better understand these functions, 3T3-L1 cell differentiation was studied by using combined proteomic and genomic strategies. The proteomics approach developed here exploits velocity gradient centrifugation as an alternative to isoelectric focusing for protein separation in the first dimension. A 20- to 30-fold increase in the concentration of numerous mitochondrial proteins was observed during adipogenesis, as determined by mass spectrometry and database correlation analysis. Light and electron microscopy confirmed a large increase in the number of mitochondrion profiles with differentiation. Furthermore, mRNA profiles obtained by using Affymetrix GeneChips revealed statistically significant increases in the expression of many nucleus-encoded mitochondrial genes during adipogenesis. Qualitative changes in mitochondrial composition also occur during adipose differentiation, as exemplified by increases in expression of proteins involved in fatty acid metabolism and of mitochondrial chaperones. Furthermore, the insulin sensitizer rosiglitazone caused striking changes in mitochondrial shape and expression of selective mitochondrial proteins. Thus, although mitochondrial biogenesis has classically been associated with brown adipocyte differentiation and thermogenesis, our results reveal that mitochondrial biogenesis and remodeling are inherent to adipose differentiation per se and are influenced by the actions of insulin sensitizers.

scholarly journals Localization of DNA in the fibrillar components of the nucleolus: a cytochemical and morphometric study.

1993 ◽  
Vol 41 (6) ◽  
pp. 829-836 ◽  
Author(s):  
M Derenzini ◽  
F Farabegoli ◽  
D Trerè
Keyword(s):  
Morphometric Analysis ◽  
Mean Value ◽  
Morphometric Study ◽  
Dense Fibrillar Component ◽  
Thin Sections ◽  
Fibrillar Center ◽  
The Mean ◽  
Fibrillar Component ◽  
Resting Lymphocytes ◽  
The Relationship

We studied the distribution of DNA in human circulating lymphocyte nucleoli using three different cytochemical methods for selective visualization of DNA in thin sections: the Feulgen-like osmium-ammine reaction, the NAMA-Ur procedure, and the osmium-ammine staining in glycine buffer, pH 1.5. All three methods indicated the presence of uniformly distributed, highly decondensed DNA filaments forming a large solitary agglomerate in the central part of the nucleolar area, corresponding to the solitary large fibrillar center (FC) as revealed by uranium and lead staining. We also studied the relationship between DNA agglomerates and nucleolar fibrillar components in resting and phytohemagglutinin (PHA)-stimulated lymphocytes by morphometric analysis of the areas occupied by these structures. In resting lymphocytes the mean area of the DNA agglomerates was 0.479 micron 2 +/- 0.161 SD, whereas that of FCs was 0.380 micron 2 +/- 0.149 SD, with a ratio of 1.26. In PHA-stimulated lymphocytes the mean area of the DNA agglomerates was 0.116 micron 2 +/- 0.056 SD, whereas that of the FCs was 0.075 micron 2 +/- 0.032 SD, with a ratio of 1.55. In PHA-stimulated lymphocytes we also measured the area occupied by the FCs plus the closely associated dense fibrillar component (DFC). The mean value of these two fibrillar components was 0.206 micron 2 +/- 0.081 SD. These data demonstrate that decondensed DNA filaments are uniformly distributed in the FCs and that in transcriptionally active nucleoli they are also present in the proximal portion of the DFC surrounding the FCs.

scholarly journals Apoptosis and Caspase-3 in Experimental Anti-Glomerular Basement Membrane Nephritis

2001 ◽  
Vol 12 (3) ◽  
pp. 485-495
Author(s):  
BIN YANG ◽  
TIMOTHY S. JOHNSON ◽  
GRAHAM L. THOMAS ◽  
PHILIP F. WATSON ◽  
BART WAGNER ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Light And Electron Microscopy ◽  
Fold Increase ◽  
Northern Blotting ◽  
Nuclear Antigen ◽  
Tubular Atrophy ◽  
Wistar Kyoto

Abstract. The caspase family is central to the proteolytic events of apoptosis. In particular, caspase-3 plays a key role in the execution of apoptosis. However, the importance of caspase-3 in renal cell apoptosis during kidney scarring has not been established. Here, nephrotoxic nephritis (NTN) was induced in Wistar Kyoto rats by a single intravenous injection of rabbit anti-rat glomerular basement membrane serum, with analysis at days 7, 15, 30, and 45 after injection. Cell apoptosis (in situ end labeling of DNA, light and electron microscopy), proliferation (proliferating cell nuclear antigen-positive cells), and inflammation (ED1-positive cells) all increased in NTN kidneys, peaking early (day 7) in the glomeruli and later (days 30 to 45) in the tubules and interstititum. The expression of caspase-3 mRNA (Northern blotting) was increased in NTN kidneys on days 7, 30, and 45 (173.3%, 228%, and 241.7%, respectively; P < 0.05). Western blotting showed that a 24-kD protein band (caspase-3 active subunit) increased with time in NTN kidneys (P < 0.01) and reached a maximum on day 45 (6.08-fold increase). A 32 kD band (caspase-3 precursor) was also increased on day 45 (3.92-fold; P < 0.01). Elevated caspase-3 activity (two- to threefold) was observed in NTN kidneys at all time points (P < 0.01). Upregulated expression of caspase-3 at all levels positively correlated with apoptosis, whereas both correlated closely with inflammation, proliferation, and subsequent fibrosis in glomeruli, tubules, and interstitium (P < 0.05). Inhibition of caspase-3 during the course of experimental nephritis may offer a new therapeutic approach for the prevention of renal apoptosis and the associated renal tubular atrophy and fibrosis.

scholarly journals Organization of the Macaque Extrastriate Visual Cortex Re-Examined Using the Principle of Spatial Continuity of Function

2011 ◽  
Vol 105 (1) ◽  
pp. 305-320 ◽  
Author(s):  
T. N. Aflalo ◽  
M.S.A. Graziano
Keyword(s):  
Functional Organization ◽  
Current Knowledge ◽  
Dimensional Space ◽  
Macaque Monkey ◽  
Specific Pattern ◽  
Extrastriate Cortex ◽  
Cortical Function ◽  
Spatial Continuity ◽  
Functional Relationships ◽  
Hierarchical Relationships

How is the macaque monkey extrastriate cortex organized? Is vision divisible into separate tasks, such as object recognition and spatial processing, each emphasized in a different anatomical stream? If so, how many streams exist? What are the hierarchical relationships among areas? The present study approached the organization of the extrastriate cortex in a novel manner. A principled relationship exists between cortical function and cortical topography. Similar functions tend to be located near each other, within the constraints of mapping a highly dimensional space of functions onto the two-dimensional space of the cortex. We used this principle to re-examine the functional organization of the extrastriate cortex given current knowledge about its topographic organization. The goal of the study was to obtain a model of the functional relationships among the visual areas, including the number of functional streams into which they are grouped, the pattern of informational overlap among the streams, and the hierarchical relationships among areas. To test each functional description, we mapped it to a model cortex according to the principle of optimal continuity and assessed whether it accurately reconstructed a version of the extrastriate topography. Of the models tested, the one that best reconstructed the topography included four functional streams rather than two, six levels of hierarchy per stream, and a specific pattern of informational overlap among streams and areas. A specific mixture of functions was predicted for each visual area. This description matched findings in the physiological literature, and provided predictions of functional relationships that have yet to be tested physiologically.

Distinguishing the sites of pre-rRNA synthesis and accumulation in Ehrlich tumor cell nucleoli

1991 ◽  
Vol 99 (4) ◽  
pp. 759-767
Author(s):  
M. Thiry ◽  
G. Goessens
Keyword(s):  
Tumor Cell ◽  
Condensed Chromatin ◽  
Rrna Genes ◽  
Rrna Synthesis ◽  
Rrna Gene ◽  
Dense Fibrillar Component ◽  
Ehrlich Tumor ◽  
Granular Component ◽  
Fibrillar Component

The precise location of transcribing rRNA genes within Ehrlich tumor cell nucleoli has been investigated using two approaches: high-resolution autoradiography of cells pulse-labelled with tritiated uridine, varying the exposure time, and in situ-in vitro transcription coupled with an immunogold labelling procedure. When autoradiographic preparations are exposed for a short time, silver grains are found associated almost exclusively with interphasic cell nucleoli. Labelling of extranucleolar areas requires longer exposure. Within the nucleolus, the first sites to be revealed are in the dense fibrillar component. Prolonging exposure increases labelling over the dense fibrillar component, with label becoming more and more apparent over the fibrillar centers. Under these conditions, however, labelling does not extend into the granular component, and no background is observed. Initiation of transcription on ultrathin cell sections occurs preferentially at the borders of condensed chromatin blocks and in their close vicinity. The condensed chromatin areas themselves remain unlabelled. Inside most nucleoli, gold-particle clusters are mainly detected in the fibrillar centers, especially at their periphery, whereas the dense fibrillar component and the granular component remain devoid of label. These results, together with previous observations made on the same cell type, clearly indicate that the fibrillar centers are the sites of rRNA gene transcription in Ehrlich tumor cell nucleoli, while the dense fibrillar component is the site of pre-rRNA accumulation.

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