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Showing papers on "Nucleolus published in 1976"


Journal ArticleDOI
TL;DR: The results support earlier reports that the expression of human ribosomal RNA (rRNA) genes is suppressed in mouse-human hybrid cells and suggest that silver staining by the Ag-AS method reflects activity of rRNA genes rather than just the presence of these genes.

428 citations


Book ChapterDOI
TL;DR: The nucleusolus is organized at the nucleolus-organizing region of the chromosomes, which are generally visible as secondary constriction regions in metaphase chromosomes, and the chromatin within the constriction region is lost at interphase inside the nucleoli.
Abstract: Publisher Summary The nucleolus is organized at the nucleolus-organizing region of the chromosomes, which are generally visible as secondary constriction regions in metaphase chromosomes. The chromatin within the constriction region is lost at interphase inside the nucleolar mass. The chromatin is highly extended at this stage. At the ultrastructural level, the nucleolus has at least three components: (1) a granular component consisting mainly of ribonucleoproteins (RNP) granules—pars granulosa, (2) a fibrillar component, consisting of RNP fibrils—pars fibrosa, and (3) chromatin elements. Chromatin elements may be present in three forms: (1) nucleolus-associated chromatin, which most likely does not take part in nucleolus formation; however, the possibility of its association with condensed inactive ribosomal cistrons, at least in some cells, cannot be overruled at present, (2) septalike intranucleolar chromatin, and (3) isolated or dispersed intranucleolar chromatin threads. Intranucleolar chromatin is often associated with the pars fibrosa. Identical components can also be found in isolated nucleoli. Studies on the nucleoli in giant chromosomes indicate that the intranucleolar chromatin in these nucleoli is present as puffs of different sizes. The nucleolar chromatin is not an autonomous structure of the nucleolus but is a continuous structure and part of the nucleolar chromosome.

372 citations


Journal ArticleDOI
01 May 1976-Cell
TL;DR: The small molecular weight RNAs of the HeLa cell have been located in specific subcellular fractions and two short-lived cytoplasmic species behave kinetically as precursors to the stable nuclear RNAs.

356 citations


Journal ArticleDOI
TL;DR: In these mouse-human hybrids, unlike those that lose human chromosomes, only humanucleolus organizer activity is expressed, and mouse nucleolus organizers activity is suppressed.
Abstract: Most mouse-human somatic cell hybrids show preferential loss of human chromosomes, absence of human 28S ribosomal RNA, and suppression of human nucleolus organizer activity, as visualized by the Ag-AS silver histochemical stain. In contrast, the mouse-human hybrids studied here show preferential loss of mouse chromosomes. The hybrids were made by fusion of HT-1080-6TG human fibrosarcoma cells with BALB/c mouse peritoneal macrophages or strain 129 mouse teratocarcinoma cells. The Ag-AS staining method shows nucleolus organizer activity of chromosomes 13, 14, 15, 21 (rarely), and 22 in the human parent and chromosomes 12, 15, 16 (rarely), and 18 in the BALB/c mouse parent. In the hybrid cells the human nucleolus organizer regions are active, as shown by Ag-AS staining and involvement in "satellite association." The mouse nucleolus organizer regions are not stained by the Ag-AS method even though mouse chromosomes 12, 15, and 18 are present in the BALB/c hybrids and at least one copy of each mouse chromosome is present in the teratocarcinoma-derived hybrids. Thus, in these mouse-human hybrids, unlike those that lose human chromosomes, only human nucleolus organizer activity is expressed, and mouse nucleolus organizer activity is suppressed.

297 citations


Journal ArticleDOI
TL;DR: A difference in the progress of nuclear changes appeared at the beginning of nuclear maturation and remained unchanged throughout the whole period studied.
Abstract: The breakdown of the pig germinal vesicle (GV) was studied in follicular oocytes matured in vivo and in vitro. For better characterization, the whole process was divided into four well-defined stages, based on the chromatin changes, and on nucleolus and nuclear membrane disappearance. In the intact germinal vesicle (GV I) nuclear membrane and nucleolus are clearly visible and chromatin forms a ring or horseshoe around the nucleolus. In the GV II a few orceinpositive structures (chromocenters) on the nuclear membrane can be detected. For the GV III slightly stained chromatin clumps, localized especially around the nucleolus, and the beginning of strand formation are typical. In the last stage - GV IV - the nuclear membrane is less distinct and the nucleolus disappears completely. Chromatin is seen as an irregular network or as individual bivalents. According to these criteria GV breakdown in vivo was completed in most oocytes between 20 to 24 hours after HCG injection. In culture a similar stage of development was reached between 16 to 20 hours. This difference in the progress of nuclear changes appeared at the beginning of nuclear maturation and remained unchanged throughout the whole period studied.

192 citations


Journal ArticleDOI
01 Mar 1976-Cell
TL;DR: The synthesis of ribosomal precursor RNA in mouse ascites nucleoli derived from cells starved for amino acids is compared with the activity of nucleoli from control cells cultivated in the presence of all amino acids and it is shown that deprivation of a single essential amino acid from the culture medium results in a drastic decrease of the RNA-forming capacity of the isolated nucleoli.

126 citations


Journal ArticleDOI
TL;DR: It is proposed that messenger precursor RNA synthesis may largely be sensitive to inhibition by DRB, and in short-term experiments, DRB has no effect on protein synthesis in L or HeLa cells.
Abstract: 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) inhibits RNA synthesis in L-929 cells (mouse fibroblast line) and HeLa cells (human epitheloid carcinoma line) within 2 min of addition of the compound to the medium. By removing DRB from the medium, the inhibition is promptly and completely reversed after treatment of cells for as long as 1 h or even longer. The inhibitory effect of DRB on the overall rate of RNA synthesis is similar in L and HeLa cells and is markedly concentration-dependent in the low dose range (5-20 muM or 1.6-6.4 mug/ml), but not as higher concentrations of DRB. At a concentration of 12 muM, DRB has a highly selective inhibitory effect on the synthesis of nuclear heterogenous RNA in L cells. At higher concentrations, there is also inhibition of 45 S ribosomal precursor RNA synthesis, but at all concentrations the effect on heterogeneous RNA synthesis in L cells in considerably greater than that on preribosomal RNA synthesis. In HeLa cells, too, DRB has a selective effect on heterogeneous RNA synthesis, but quantitatively the selectivity of action is somewhat less pronounced. In both L and HeLa cells, the inhibition of synthesis of nuclear heterogeneous RNA is incomplete even at very high concentrations of DRB (150 muM). Thus, while DRB is a selective inhibitor of nuclear heterogeneous RNA synthesis, not all such RNA synthesis is sensitive to inhibition. It is proposed that messenger precursor RNA synthesis may largely be sensitive to inhibition by DRB. In short-term experiments, DRB has no effect on protein synthesis in L or HeLa cells. DRB has a slight to moderate inhibitory effect on uridine uptake into L cells and a moderate to marked effect on uptake of uridine into HeLa cells.

107 citations


Journal ArticleDOI
TL;DR: In this paper, high-resolution autoradiography using [3H]actinomycin D or [3h]TdR reveals that a small amount of DNA is present inside the fibrillar centres.

91 citations


Journal ArticleDOI
TL;DR: The results show that the pattern of transcriptional control of rRNA genes differs widely in different genes, even in the same genetic unit, and that rRNA synthesis is regulated primarily at the level of transcription.
Abstract: Natural changes in the transcription of rRNA genes were studied in nucleoli from three oogenic stages of the newt Triturus alpestris with electron microscope, auto-radiographic, and biochemical techniques From determinations of the uridine triphosphate pool sizes and [3H]uridine uptake, phosphorylation, and incorporation into 28S and 18S rRNAs in vivo it was estimated that the rate of rRNA synthesis was about 001% in previtellogenic oocytes and 13% in mature oocytes when compared to midvitellogenesis Spread preparations of nucleoli showed significant morphological changes in the transcriptional complexes The total number of lateral fibrils, ie, ribonucleoproteins containing the nascent rRNA precursor, were drastically decreased in stages of reduced synthetic activity This indicates that rRNA synthesis is regulated primarily at the level of transcription The resulting patterns of fibril coverage of the nucleolar chromatin axes revealed a marked heterogeneity On the same nucleolar axis occurred matrix units that were completely devoid of lateral fibrils, matrix units that were almost fully covered with lateral fibrils, and various forms of matrix units with a range of lateral fibril densities intermediate between the two extremes Granular particles that were tentatively identified as RNA polymerase molecules were not restricted to the transciptional complexes They were observed, although less regularly and separated by greater distances, in untranscribed spacer regions as well as in untranscribed gene intercepts The results show that the pattern of transcriptional control of rRNA genes differs widely in different genes, even in the same genetic unit

84 citations


Journal Article
TL;DR: Nucleoli of rat myocardial cells did not recover but underwent further fragmentation, segragation, and conversion to ring-shaped structures, which may represent an important aspect of adriamycin toxicity.
Abstract: Ultrastructural studies of the effects of adriamycin on liver and cardiac cell nucleoli of the rat showed that nucleolar segregation occurred within 1 hr after an i.v. injection of a 40-mg/kg dose. Between 3 and 27 hr after this single dose, liver cell nucleoli progressively reverted to a normal ultrastructure. Nucleoli of rat myocardial cells did not recover but underwent further fragmentation, segregation, and conversion to ring-shaped structures. The ultrastructural alterations of myocardial cell nucleoli may represent an important aspect of adriamycin toxicity.

58 citations


Journal Article
TL;DR: Injected nuclei appear to increase their rate of RNA synthesis during incubation in oocytes, correlated with nuclear enlargement, as well as with the loss of protein from injected nuclei and with their uptake of histone and nonhistone proteins from oocyte cytoplasm.
Abstract: Nuclei from HeLa and other mammalian cells have been injected into Xenopus oocytes. The synthesis, uptake, and release of RNA and proteins by injected nuclei have been investigated by autoradiography. Injected nuclei which undergo enlargement synthesize RNA continuously for up to 28 days. When oocytes are incubated in [3H]uridine or [3H]guanosine, injected nuclei are labelled nearly as strongly as the nucleoli, but much more strongly than the nucleoplasm of the oocyte's germinal vesicle. Injected nuclei appear to increase their rate of RNA synthesis during incubation in oocytes. This apparent increase in the rate of RNA synthesis is correlated with nuclear enlargement, as well as with the loss of protein from injected nuclei and with their uptake of histone and nonhistone proteins from oocyte cytoplasm. Injected HeLa nuclei lose most of the previously synthesized RNA from their nucleoplasm, but little if any of the RNA from their remaining nucleoli.

Journal ArticleDOI
TL;DR: In situ hybridization using tritium-labeled 28S and 18S RNA shows that in the interphase nucleus the acrocentric short arms, carriers of ribosomal cistrons, are associated with the nucleolus, confirming that theucleolus constitutes one of the orientation points determining the relative localization of chromosomes in theinterphase nucleus.
Abstract: Use of specific stains permits analysis of the frequency of nucleolus-associated heterochromatin in chromosomes 1 and 9 from human fibroblasts. In 81% of interphase nuclei the heterochromatic segment of both No. 1 chromosomes is associated with the nucleolus, while in 19% only one heterochromatic segment shows such an association with the other occupying a random position in the nucleoplasm. The nucleolar association of chromosome 9 heterochromatin is less constant: in 42.3% of the nuclei both segments are associated with the nucleolus, in 39% of the nuclei only one heterochromatic segment presents such an association, and in 18.7% neither of the two heterochromatic segments is in nucleolar association. In 6% of the cells, one or two chromosome 9 heterochromatic segments are in contact with the nuclear membrane.

Journal ArticleDOI
TL;DR: The localization of nucleolar organizers in the quail oocyte was studied during meiotic prophase I and no decisive argument exists excluding either the juxtanucleolar knob, which appears to be formed of facultative heterochromatin, or the DNP fibers of the nucleolonema.

Journal ArticleDOI
TL;DR: The onset of RNA synthesis in primary roots of germinating Zea mays embryos was studied and it was observed that there are three successive stages in the reactivation ofRNA synthesis.
Abstract: The onset of RNA synthesis in primary roots of germinating Zea mays embryos was studied. Incubation of excised embryos in the radioactive precursor solution was performed after different germination periods.It was observed that there are three successive stages in the reactivation of RNA synthesis. Stage one is characterized by the synthesis in the chromatin of heterodisperse nuclear RNA rich in adenine. Stage two is characterized by the synthesis of heterodisperse nuclear RNA in the chromatin and 2.3 x 10(6) rRNA precursor in the nucleolus. Stage three is characterized by the synthesis of heterodisperse nuclear RNA and rRNA precursor and by the processing of the latter in mature rRNA.

Journal ArticleDOI
TL;DR: All but a few proteins exhibited relative nucleolar stoichiometry values of approximately one, indicating that there are not significant pools of most ribosomal proteins in isolated nucleoli.

Journal ArticleDOI
TL;DR: As the number of phases of the DNA cycle increases during the first 48 h of lymphocyte culture, there is a corresponding increase in the variety of nucleolar morphology and some of the new nucleoli differentiating in post-telophase show processes of RNP from their surfaces.

Journal ArticleDOI
TL;DR: Results show that syntheses of individual low molecular weight nuclear (LMWN) RNAs are controlled by different regulatory mechanisms, and there appears to be a specific relationship between U3 RNA and functional states of the nucleolus.
Abstract: Studies were made on the effects of alpha-amanitin, cycloheximide, and thioacetamide on synthesis and content of low molecular weight nuclear RNA. Cycloheximide, an inhibitor of protein synthesis and the synthesis of 45S pre-rRNA and 5S RNA, also inhibited synthesis of nuclear U1 and U3 RNAs. alpha-Amanitin, an inhibited the synthesis of U1 and U2 low molecular weight nuclear RNA. Thioacetamide, which induces nucleolar hypertrophy and increased nucleolar RNA polymerase activity, markedly increased synthesis of 5.8S RNA and U3 RNA. These results show that syntheses of individual low molecular weight nuclear (LMWN) RNAs are controlled by different regulatory mechanisms. In particular, there appears to be a specific relationship between U3 RNA and functional states of the nucleolus.

Journal ArticleDOI
TL;DR: A new nucleolar fraction that is observed following drug treatment of tissue culture cells has been identified and its properties are described and evidence which suggests that microspherules represent a condensed form of chromatin has been presented.

Journal ArticleDOI
TL;DR: The results strongly suggest that the above-mentioned procedure is actually fractionating RNA polymerase I into free and bound forms, and that IA form is not an artifact of isolation but is always present in vivo at a certain amount, although the exact nature of this molecule is not known at present.
Abstract: RNA polymerase I (or A) was extracted from nuclear, nucleolar and nucleoplasmic fractions, and resolved into IA and IB forms on a phosphocellulose column. During the course of cyclo-heximide treatment, the activity of RNA polymerase IB decreased in the nucleoli with concomitant increase in the nucleoplasmic fraction, suggesting strongly that cycloheximide induced specific leakage of IB enzyme from the nucleolus. The activity of IA enzyme did not change in the nucleoli. When nucleoli were incubated in the presence of actinomycin D, all the IA enzyme activity and approximately 30% of the total IB enzyme activity were released in the incubation medium, whereas 70% of IB activity remained associated with the nucleolar pellet where no IA activity was detected. The enzyme which was released into the incubation medium was tentatively designated as free or unbound RNA polymerase I and that which was associated with the nucleolar pellet as template-bound enzyme. During the treatment with cycloheximide, the activity of bound enzyme, which contained exclusively IB form, decreased rapidly, with kinetics almost identical to that of nucleolar RNA synthesis in vivo. The activity of free enzyme did not change appreciably. At 2 h after partial hepatectomy, IB enzyme activity in the free RNA polymerase fraction increased to almost twice the control, while the bound enzyme activity did not increase appreciably until 4 h of regeneration. Enhancement of nucleolar RNA synthesis in vivo was not apparent at 2 h but became significant by 4 h after partial hepatectomy. These results strongly suggest that (a) the above-mentioned procedure is actually fractionating RNA polymerase I into free and bound forms, (b) RNA polymerase IB is the transcriptionally active form in vivo, (c) RNA polymerase IB exists in excess in the nucleoli, but the amount of bound IB molecules, which are engaged in transcription in vivo, must be determined by some other factor(s) than the mere concentration of IB enzyme in the nucleolus, and (d) IA form is not an artifact of isolation but is always present in vivo at a certain amount, although the exact nature of this molecule is not known at present.

Journal ArticleDOI
TL;DR: The results strongly suggest that the above-mentioned procedure is actually fractionating RNA polymerase I into free and bound forms, and that IA form is not an artifact of isolation but is always present in vivo at a certain amount, although the exact nature of this molecule is not known at present.
Abstract: RNA polymerase I (or A) was extracted from nuclear, nucleolar and nucleoplasmic fractions, and resolved into IA and IB forms on a phosphocellulose column. During the course of cycloheximide treatment, the activity of RNA polymerase IB decreased in the nucleoli with concomitant increase in the nucleoplasmic fraction, suggesting strongly that cycloheximide induced specific leakage of IB enzyme from the nucleolus. The activity of IA enzyme did not change in the nucleoli. When nucleoli were incubated in the presence of actinomycin D, all the IA enzyme activity and approximately 30% of the total IB enzyme activity were released in the incubation medium, whereaa 70% of IB activity remained associated with the nucleolar pellet where no IA activity was detected. The enzyme which was released into the incubation medium was tentatively designated as free or unbound RNA polymerase I and that which was associated with the nucleolar pellet as template-bound enzyme. During the treatment with cycloheximide, the activity of bound enzyme, which contained exclusively IB form, decreased rapidly, with kinetics almost identical to that of nucleolar RNA synthesis in vivo. The activity of free enzyme did not change appreciably. At 2 h after partial hepatectomy, IB enzyme activity in the free RNA polymerase fraction increased to almost twice the control, while the bound enzyme activity did not increase appreciably until 4h of regeneration. Enhancement of nucleolar RNA synthesis in vivo was not apparent at 2 h but became significant by 4 h after partial hepatectomy. These results strongly suggest that (a) the above-mentioned procedure is actually fractionating RNA polymerase I into free and bound forms, (b) RNA polymerase IB is the transcriptionally active form in vivo, (c) RNA polymerase IB exists in excess in the nucleoli, but the amount of bound IB molecules, which are engaged in transcription in vivo, must be determined by some other factor(s) than the mere concentration of IB enzyme in the nucleolus, and (d) IA form is not an artifact of isolation but is always present in vivo at a certain amount, although the exact nature of this molecule is not known at present.

Journal ArticleDOI
TL;DR: Results are interpreted to indicate that rRNAmethylases are regulated by cellular metabolites during the nucleolar biosynthesis of ribosomes and that r RNA methylases may provide a favorable site for selective action by cancer chemotherapeutic agents.
Abstract: The activity of rRNA methylases was stimulated by high-energy precursors of RNA (ribonucleoside triphosphates) and inhibited by degradation products of RNA (ribonucleotides and oligoribonucleotides) The response of methylases from rat Novikoff ascites tumor and liver to these metabolites was strikingly different The highly active tumor enzymes responded preferentially to inhibition by catabolic metabolites, whereas the less active liver enzymes responded exclusively to stimulation by anabolic metabolites When the activity of rRNA methylases was assayed in response to increasing concentration of S-adenosylmethionine, the tumor enzymes responded with a hyperbolic substrate dependence curve and the liver enzymes with a sigmoidal curve In the presence of an inhibitory dinucleotide, ApA, the tumor enzymes responded with a sigmoidal curve; in the presence of a stimulator, adenosine 5'-triphosphate, the liver enzymes responded with a hyperbolic substrate concentration curve When normal rats were subject to a series of treatments by thioacetamide, a hepatocarcinogen, the liver nucleolar rRNA methylases became responsive to inhibition by ApA and relatively unresponsive to stimulation by adenosine 5'-triphosphate When tumor-bearing rats were treated with polyinosinate:polycytidylate, an antitumor agent, the tumor nucleolar rRNA methylases became unresponsive to inhibition by ApA and more responsive to stimulation by adenosine 5'-triphosphate A correlation was noted between increased methylation efficiency in vivo and increased stability of nucleolar RNA during incubation in vitro, or vice versa These results are interpreted to indicate that rRNAmethylases are regulated by cellular metabolites during the nucleolar biosynthesis of ribosomes and that rRNA methylases may provide a favorable site for selective action by cancer chemotherapeutic agents

Journal ArticleDOI
TL;DR: Isolated rat hepatic nuclei were shown to contain poly(A) polymerase in two distinct physiologically active forms, one of which was associated with the chromatin fraction and was dependent on endogenous RNA, presumably mRNA and the other was localized in the nuclear sap in a 'free' form and was stimulated almost 30-40-fold by exogenously added poly( A).
Abstract: Isolated rat hepatic nuclei were shown to contain poly(A) polymerase in two distinct physiologically active forms. One form was associated with the chromatin fraction and was dependent on endogenous RNA, presumably mRNA. The other activity was localized in the nuclear sap in a 'free' form and was stimulated almost 30-40-fold by exogenously added poly(A). Isolated nucleoli were devoid of significant poly(A)-synthesizing activity.

Journal Article
TL;DR: Stages of meiosis from the bluebell Endymion non-scriptus (L.) were studied by electron microscopy and demonstrated that the pale-staining region of nucleoli is the nucleolus organizer and almost certainly the chromosome region containing the ribosomal cistrons.
Abstract: Stages of meiosis from the bluebell Endymion non-scriptus (L.) were studied by electron microscopy. The nucleolus went through the process of segregation at the beginning of meiosis with the movement to its surface of a pale-staining region. This region was shown to be the same as that called the 'L zone' or lacunae of nucleoli. Its chromosomal nature was strongly suggested by the presence of the synaptonemal complex within it. This demonstrated that the pale-staining region of nucleoli is the nucleolus organizer and almost certainly the chromosome region containing the ribosomal cistrons, and justifies the use of these terms to describe the structure when seen inside the nucleolus. The relationship between this zone and the heterochromatic knob called the nucleolar organizing body in maize by other workers is discussed.

Journal ArticleDOI
TL;DR: Analysis of the effect of translational inhibitors on mouse embryo cells of different genotypes indicates that activation of viral RNA is specific for one endogenous virus class and is a dominant genetic characteristic.
Abstract: Several biologically distinguishable type-C RNA viruses are genetically transmitted in mouse cells. In the present report, chemicals that inhibit several different steps in protein synthesis are shown to cause marked increases in the cellular concentration of virus-specific RNA and the subsequent induction of virus. Analysis of the effect of translational inhibitors on mouse embryo cells of different genotypes indicates that activation of viral RNA is specific for one endogenous virus class and is a dominant genetic characteristic. Two lines of evidence favor the hypothesis that the induction of viral RNA involves transcriptional derepression rather than an alteration in its post-transcriptional processing. First, nuclear and cytoplasmic fractions of induced cells are shown to demonstrate similar increases in their concentrations of virus-specific RNA. Second, the decay of induced viral RNA following inhibition of further RNA synthesis by actinomycin D is not prevented by continued exposure to the inducer. These findings weigh heavily against the possibility that translational inhibitors act to stabilize viral RNA post-transcriptionally. The results are consistent with a model in which the expression of one class of endogenous virus is regulated by a labile repressor protein acting at a transcriptional level.

Journal ArticleDOI
TL;DR: Allium cepa L. root meristems, after a brief fixing in cold methanol, retain endogenous RNA polymerase activity, supplying a new evidence in favour that prenucleolar bodies are not synthesized in telophase since they appear before any transcription reinitiates.

Journal ArticleDOI
TL;DR: Evolution of the nucleoli has been followed during oogenesis in theAcipenserid fishes, Acipenser ruthenus (the sterlet) and A. guidenstadti (the sturgeon) using light and electron microscopes.
Abstract: Evolution of the nucleoli has been followed during oogenesis in the Acipenserid fishes, Acipenser ruthenus (the sterlet) and A. guldenstadti (the sturgeon) using light and electron microscopes. In the ovaries of adults, the oogonial nuclei usually have a single nucleolus with an adjacent mass of paranucleolar fibrillar material. The cytoplasm of the oogonia contains two dense bodies peculiar only to gonocytes, one being electron dense and containing RNA and the other being electron-lucent and lacking RNA. Neither is surrounded by membrane. The fine structure of the electron-lucent body is identical to that of the paranucleolar material, while the RNA-containing body structurally resembles the nucleolus. A nuclear origin forboth cytoplasmic bodies is likely. Leptoten-stage oocytes usually still have a single nucleolus. During zygotene, it is adjacentto the nuclear envelope and opposite to the chromosomes contracted in synizesis. At pachy-tene, a ‘cap’ of extra chromosomal chromatin is formed under the nuclear envelope and around the nucleolus. Bivalents also contact this cap. In early diplotene, the primary nucleolus still persists. The material of the cap is dispersed beneath the entire nuclear envelope in the form of granules of extra DNA; each granule then produces a peripheral (secondary) nucleolus. These become typical amphinucleoli with differentially developed granular parts, depending on the age of the nucleolus and the stage of meiosis. Their fibrillar parts always face the nuclear envelope. New peripheral nucleoli continue to form as long as granules of extra DNA persist under the nuclear envelope, i.e. approximately until vitello-genesis. In early vitello-genesis, the peripheral nucleoli become transformed, by re-distribution of their fine structural components, into circular threads trailing towards the centre of the nucleus. The axis of the thread consists of fibres and is coated with granules. In late vitello-genesis, the nucleoli round up and become vacuolized; they are then peripheral again. Proteinaceous RNA-lacking structures which are also produced in the nuclei during oogenesis in the A cipenseridae, are the ‘nuclear bodies’ and ‘spheres’. The former are adjacent to peripheral nucleoli, the latter form on lampbrush chromosomes. Both are ultra-structurally alike. The loops of lamp brush chromosomes produce also RNP bodies (‘granules’ of Callan & Lloyd, 1960) which are ultra structurally similar to nucleoli but lack segregation of granules from fibres into spatially distinct parts. The evolution of the nucleolar apparatus during oogenesis in the Acipenseridae closely resembles that in amphibians.

Journal ArticleDOI
TL;DR: The relationship of ribosomal RNA (rRNA) synthesis to nucleolar ultrastructure was studied in partial nucleolar mutants of Xenopus laevis and demonstrated that nucleolar segragation is related specifically to a reduction in rRNA synthesis.
Abstract: The relationship of ribosomal RNA (rRNA) synthesis to nucleolar ultrastructure was studied in partial nucleolar mutants of Xenopus laevis. These mutations are the result of a partial deletion of rRNA genes and therefore alow studies on nucleolar structure and function without using drugs that inhibit rRNA synthesis. Ultrastructural studies demonstrated that normal embryos have reticulated nucleoli that are composed of a loose meshwork of granules and fibrils and a typical nucleolonema. In contrast, partial nucleolar mutants in which rRNA synthesis is reduced to less than 50% of the normal rate have compact nucleoli and nucleolus-like bodies. The compace nucleoli contain granules and fibrils, but they are segregated into distinct regions, and a nucleolonema is never seen. Since other species of RNA are synthesized normally by partial nucleolar mutants, these results demonstrate that nucleolar segragation is related specifically to a reduction in rRNA synthesis. The nucleolus-like bodies are composed mainly of fibrils,and the number of such bodies are composed mainly of fibrils, and the number of such bodies present in the different nucleolar mutants is inversely related to the relative rate of rRNA synthesis. Although the partial nucleolar organizers produce segregated nucleoli in these mutants, they organize morphologically normal, but smaller, nucleoli in heterozygous embryos. Alternative explanations to account for these results are discussed.

Journal ArticleDOI
TL;DR: Nucleoli were studied in chick fibroblasts cultured in vitro, under normal or under experimental conditions, and in several mammalian cell types in vivo and it appears that they could be related to the nucleolar organizers as suggested in Ehrlich tumours.
Abstract: Nucleoli were studied in chick fibroblasts cultured in vitro, under normal or under experimental conditions, and in several mammalian cell types in vivo. All these cells frequently contain nucleoli with fibrillar centres. The nucleolar fibrillar centres are composed of fibrous material of low electron density and are always intimately associated with the dense fibrillar component. Their morphology is very similar to that analysed cytochemically in Ehrlich tumour cells. It therefore appears that they could be related to the nucleolar organizers as suggested in Ehrlich tumour cells.

Journal ArticleDOI
TL;DR: The nucleolar proteins separated by SDS gel electrophoresis have been examined throughout the growth cycle of Acanthamoeba and into the strat of induced encystment, leaving the observation that extremely few protein homologies exist between the two.
Abstract: A technique is described for isolating nuceoli from Acanthamoeba castellanii. Nuclei isolated by a modification of the technique of F. J. Chlapowski and R. N. Band (1971) are sonicated in a surcrose-Tris-MgSO4-KC1-Triton X-100 buffer and centrifuged on a linear sucrose gradient extending from 1.3 M to 1.5 M with a 2.6 M cushion, at 41000 rpm for 90 min. The only apparent contaminants in the nucleolar preparation are outer cyst walls. A procedure is described for the isolation of chemically pure outer cyst walls, and a comparison of the proteins with the nucleolar preparation reveals that outer cyst walls represent negligible contaminants. The ultrastructure of these isolated nucleoli examined with transmission electron microscopy is found to be identical with that of nucleoli from whole cells, fixed in an identical manner. The 50 nucleolar proteins separated by SDS gel electrophoresis have been examined throughout the growth cycle of Acanthamoeba and into the strat of induced encystment, at which time 10 protein bands disappear, 11 bands are observed to decrease, and 8 are seen to increase in concentration. Phenol-soluble proteins are extracted from the nucleolus which correspond to 29 of the 50 nucleolar proteins, with 17 of these proteins corresponding to nucleolar proteins that change at the onset of encystment. Thes nucleolar proteins are also compared with those of rat liver nucleoli by gel electrophoresis, resulting in the observation that extremely few protein homologies exist between the two. Numerous quantitative and qualitative changes in the gel pattern of phenol-soluble nuclear proteins during early and late log phase growth and the onset of stationary phase were also observed.

Journal ArticleDOI
TL;DR: Observations on stain uptake by thin sections through condensed interphase chromosomes in cells from epithelial and muscle tissue in kidney and intestine, and also in fibroblasts, show a distribution into DNA-rich and DNA-poor phases similar to that already described in Cells from the connective tissue blood.
Abstract: Observations on stain uptake by thin sections through condensed interphase chromosomes in cells from epithelial and muscle tissue in kidney and intestine, and also in fibroblasts, show a distribution into DNA-rich and DNA-poor phases similar to that already described in cells from the connective tissue blood. In all the nuclei the nucleolus, when adjacent to the nuclear envelope, is separated from the inner membrane by a monolayer of chromatin structural units, similar to the monolayer enclosed on both sides by nuclear envelope, previously described in a wide variety of organisms. The data provide further support for the hypothesis that the condensed interphase chromosomes in eukaryotes are characterized by essentially similar structural units folded to form similar patterns. This hypothesis, regarding the higher order units, is consistent with data of others which show that histones and DNA fold to form similar repeating subunits in chromatin, irrespective of the base sequence in the DNA and the origin of the histones.