Cytological evidence on the ability of the nucleolus organizing regions to assemble pre-existing nucleolar material

Summary The treatment ofVicia faba root tips with 5-azacytidine caused unequal allotment of the chromosomes; after one cell cycle, a pair of daughter nuclei was produced, one of which carried the nucleolus organizing regions (NORs) and the other of which lacked them. The pre-existing nucleolar material brought into the NOR-carrying daughter nucleus by the chromosomes disappeared concurrently with the appearance of the new nucleoli, whereas in another daughter nucleus without the NOR a number of small nucleolus-like bodies became visible at random positions. These findings suggest that in addition to ribosomal RNA synthesis the NOR has another role in cells; to stimulate the pre-existing nucleolar material to assemble at the NOR itself.     

Detection of nucleolus organizer regions (NOR) in the chromosomes of the domestic pig (Sus scrofa domestica L.)

Summary In the domestic pig (Sus scrofa domestica L.) nucleolar organizer regions (NOR) were detected by a combined silver-Giemsa method (Ag-G). The main sites of NORs are the secondary constrictions of chromosomes 8 and 10. Sometimes an additional NOR was observed near the centromere of 1 homologue of chromosome 11. Associations of NORs were seen only between chromosomes 10 and at a very low frequency.     

Detection of nucleolus organizer regions (NOR) in the chromosomes of the domestic pig (Sus scrofa domestica L.)

In the domestic pig (Sus scrofa domestica L.) nucleolar organizer regions (NOR) were detected by a combined silver-Giemsa method (Ag-G). The main sites of NORs are the secondary constrictions of chromosomes 8 and 10. Sometimes an additional NOR was observed near the centromere of 1 homologue of chromosome 11. Association of NORs were seen only between chromosomes 10 and at a very low frequency.     

The silver-staining technique—a tool for characterizing lymphocyte populations in mammalian peripheral blood

Summary Nucleolus organizer regions (NORs) of the peripheral blood lymphocytes (PBL) of 9 mammalian species were analyzed by means of a silver-staining procedure. Species-specific NOR patterns were demonstrated. The number of NOR chromosomes was positively correlated with the nucleolar coefficient, and negatively correlated with the relative frequencies of uninucleolar cells in PBL interphase.Acknowledgment. The authors thank Professor Dr E. Groll-Knapp, of the Institute of Environmental Hygiene at the University of Vienna, for statistically evaluating the data provided.     

Position of the nucleolus within the nuclei of pachytene spermatocytes of Dromiciops australis and Marmosa elegans (Didelphoidea-Marsupialia).

The location of the nucleoli within the nuclei of pachytene spermatocytes, and their relation with the position of the nucleolar organizer region (NOR) was studied. It appears that a terminal NOR determines a peripheral location of the nucleolus, due to the position of the NOR over the synaptonemal complex and to the attachment of the nucleolar chromosome telomeres at the nuclear membrane.     

Telomere dynamics unique to meiotic prophase: formation and significance of the bouquet

Telomeres carry out conserved and possibly ancient functions in meiosis. During the specialized prophase of meiosis I, meiotic prophase, telomeres cluster on the nuclear envelope and move the diploid genetic material around within the nucleus so that homologous chromosomes can align two by two and efficiently recombine with precision. This recombination is in turn required for proper segregation of the homologs into viable haploid daughter cells. The meiosis-specific telomere clustering on the nuclear envelope defines the bouquet stage, so named for its resemblance to the stems from a bouquet of cut flowers. Here, a comparative analysis of the literature on meiotic telomeres from a variety of different species illustrates that the bouquet is nearly universal among life cycles with sexual reproduction. The bouquet has been well documented for over 100 years, but our understanding of how it forms and how it functions has only recently begun to increase. Early and recent observations document the timing and provide clues about the functional significance of these striking telomere movements.     

Chromosome arrangement throughout mitosis and interphase inAllium sativum (Liliaceae)

Summary Allium sativum (garlic) root-tip chromosomes were subjected to a C-banding procedure. In addition to the nucleolar bands reported previously in this species, bands which are telomeric or close to the telomeres have been detected in some pairs. This has allowed us to analyze the arrangement of chromosomes during interphase.     

Chromatin structure contribution to the synaptonemal complex formation

Meiosis is a key cellular and molecular process for sexual reproduction contributing to the genetic variability of organisms. This process takes place after DNA replication and consists in a double cellular division, giving rise to four haploid daughter cells or gametes. Meiotic recombination between homologous chromosomes, in the meiotic prophase I, is mediated by a tripartite structure named Synaptonemal Complex (SC). The SC is a peptidic scaffold in which the chromatin of homologous chromosomes is organized during the pachytene stage, holding chromosomes together until the meiotic recombination and genetic exchange have taken place. The role of chromatin structure in formation of the SC and the meiotic recombination at meiotic prophase I remain largely unknown. In this review we address the epigenome contribution to the SC formation at meiotic prophase I, with particular attention on the chromatin structure modifications occurring during the sub-stages of meiotic prophase I. Received 18 September 2008; received after revision 10 October 2008; accepted 24 October 2008     

Fine structure of differentiated chromosome segments during the meiosis of pollen mother cells of Allium cepa

Summary Differentiated chromosome segments were observed during studies of pollen mother cells of A. cepa during the first meiotic division. Their structure is like those of the nucleolar organizing region (NOR) described in A. cepa microspores. It is suggested that these differentiated chromosome segments correspond to the secondary constrictions seen under the optical microscope.     

Distribuzione di fosfatasi alcaline in gonadi di Crostacei isopodi in rapporto al ciclo degli acidi nucleici

Summary The distribution of alkaline phosphatases has been studied in male and female gonads of some isopod crustaceans by means of Gomori's reaction.Monophosphatases are found on all Feulgen-positive nuclear structures, in the nucleolus and in the cytoplasm of young oocytes. They disappear from the cytoplasm in auxocytosis, during yolk formation, although they persist in the germinal vesicle. The authors think that the cytoplasmic localization is coincident with ribonucleic acid distribution.From the experiments related in the following note, nucleolar phosphatases appear to be in some way different from the nuclear (sensu stricto) and cytoplasmic phosphatases.Diphosphatases do not follow so closely the nucleic acid localization. They are absent from cytoplasm; in the nucleolus they are not distributed uniformly but show characteristic structure; in the nucleus they are not localized in the chromosomes but homogeneously diffused.     

Nucleolar localization and circadian regulation of Per2S,a novel splicing variant of the Period 2 gene

In this work, we show for the first time that a second splicing variant of the core clock gene Period 2 (Per2), Per2S, is expressed at both the mRNA and protein levels in human keratinocytes and that it localizes in the nucleoli. Moreover, we show that a reversible perturbation of the nucleolar structure acts as a resetting stimulus for the cellular clock. Per2S expression and periodic oscillation upon dexamethasone treatment were assessed by qRT-PCR using specific primers. Western blot (WB) analysis using an antibody against the recombinant human PER2 (abRc) displayed an intense band at a molecular weight of ~55 kDa, close to the predicted size of Per2S, and a weaker band at the expected size of Per2 (~140 kDa). The antibody raised against PER2 pS662 (abS662), an epitope absent in PER2S, detected only the higher band. Immunolocalization studies with abRc revealed a peculiar nucleolar signal colocalizing with the nucleolar marker nucleophosmin, whereas with abS662 the signal was predominantly diffuse all over the nucleus and partially colocalized with abRc in the nucleolus. The analysis of cell fractions by WB confirmed the enrichment of PER2S and the presence of PER2 in the nucleolar compartment. Finally, a pulse (1 h) of actinomycin D (0.01 μg/ml) induced reversible nucleolar disruption, PER2S de-localization and circadian synchronization of clock and Per2S genes. Our work represents the first evidence that the Per2S splicing isoform is a clock component expressed in human cells localizing in the nucleolus. These results suggest a critical role for the nucleolus in the process of circadian synchronization in human keratinocytes.     

Variable positions of nucleolus organizer regions in Bovidae

Summary Silver NOR staining has been applied to cattle (Bos taurus L.), goat (Capra hircus L.) and sheep (Ovis aries L.) chromosomes. The sites of silver NORs showed variation within the family Bovidae probably due to a reciprocal translocation event.     

The karyotypes of the Corsican and Sardinian mountain salamanders,Euproctus montanus andE. platycephalus (Urodela: Salamandridae)

Summary The chromosomes ofEuproctus montanus andE. platycephalus were studied by means of the C-banding method and the AS-SAT technique which are useful for identifying the single pairs of the complement and for recognizing nucleolar organizer regions. According to the morpho-structural characteristics shown by the specific karyotypes, it has been possible to draw some cytotaxonomic deductions concerning the karyological evolution within the insular group.This work was supported by grant CT 76.01271.04/115.3822 from C.N.R., Rome.     

Constitutive heterochromatin and nucleolus organizer region in the knifefish,Apteronotus albifrons (Pisces,Apteronotidae)

Summary The C-banding and silver staining of the chromosomes of the knifefishApteronotus albifrons (2n=24), demonstrated the presence of constitutive heterochromatin in the centromeric region of every chromosome, except pair 4, where the entire long arm was darkly stained, the silver stain positive nucleolus organizer region (NOR) being embedded in it.The authors are grateful to Dr Sen Pathak (University of Texas System Cancer Center at Houston, Texas, USA) and Dr Yatiyo Yassuda (Universidade de São Paulo, São Paulo, Brasil) for their suggestions. This study was supported in part by grants of the Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq).     

Observations on the dynamics of argyrophilic nucleolar material in the nuclei of mice spermatids

Applying a new silver staining technique it could be shown that in very early spermatids strong argyrophilia in nucleoli is confined to their granular and fibrillar components; fibrillar centers are devoid of silver. During subsequent developmental stages remnants of these nucleolar components are present in the form of intensively silver stained clusters of coiled fibers. As chromatin condensation proceeds, these fibrous structures decrease in size and density and are finally completely absent. The nucleus of the mature sperm contains only the space in which they formerly existed, now silver negative, as the so-called 'nuclear vacuole'.     

Bau und Funktion der Nukleolen

Summary In previous years the composition and function of chromosomes have been analyzed to a considerable extent, in contrast to the small number of investigations that have been made on nucleoli. There exist at present many conflicting opinions about the structure and function of the nucleolus, and therefore a critical survey of the facts and theories about the nucleolar substance appears necessary.The structure, chemical composition and metabolism of the nucleoli of various cells are described in this publication. The regulation of the size and function of the nucleolus is also discussed.It is concluded that the size and structure of the nucleolus is organ- and species-specific. A rapid synthesis of RNA and proteins occurs in the nucleolus. After a short interval, this disappears; and it is therefore concluded that the nucleolus must have a high turnover of these substances. The size and function of the nucleoli depends on the energy state of the cytoplasm.It appears that the nucleolus can synthesize proteins which can be utilized by the chromosomes during their genetic function.     

Etude de la consommation d'oxygène et de la teneur en acide désoxyribonucléique du foie à divers âges chez le rat

Summary The quantitative study of whole liver DNA and liver nuclei DNA at various ages, and the simultaneous study of oxygen uptake shows the following facts:(1) The rate of oxygen consumption to the fresh weight or to the DNA or to diploid chromosomes group is the same for the adult or the young aged 8 days. The values of oxygen consumption calculated for one cell or for one nucleus (taking notice of the fact that the mean level of liver nuclei DNA varies with the age) at 8 days is only the half of the adult value. Even at two months the value is 20% lower than in the adult.(2) In older animals there is a decrease of oxygen uptake which is more noticeable on a cellular scale than with the classical rate to fresh or dry weight. We affirm the fact that the diminution of the oxygen uptake of each cell is compensated by the increase of the number of cells.     

An ammoniacal silver staining technique for mitotic chromosomes ofTriturus (Urodela: Salamandridae)

Summary An ammoniacal silver staining technique was applied to mitotic metaphase chromosomes of 2 species of Newts (Triturus). The method is useful for identifying nucleolar organizer regions. In addition, it reveals other sites of unknown significance.With financial support of the Consiglio Nazionale delle Ricerche, Rome.     

Nuclear lamellae in the germ-line cells of gall midges (Cecidomyiidae,Diptera)

Summary Lamellar structures in oogonial and spermatogonial cells of gall midges were found to form a complicated system of intranuclear compartments inside which all heteropycnotic chromosomes present in the germ-line cells of these insects are located. In this way, the heteropycnotic S-chromosomes are separated by the nuclear lamellae from the remaining, decondensed chromosomes (E-chromosomes) of the interphase germ-line nucleus.This investigation was supported in part under Contract DPKBN/52/76-II.1.3.10. with the Polish Academy of Sciences.