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.     

Larvale und imaginale Mitteldarmchromosomen vonDrosophila repleta

Summary In certain tissues ofD. repleta-flies, it is possible to observe the interphase chromosomes that are either of larval origin or that occur in imaginal tissues. The imaginal interphase chromosomes from the midgut are similar to the small larval midgut chromosomes in the early phases of their development.     

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.     

A simple one-step procedure for staining the nucleolus organizer regions.

A simple silver staining technique for routine use is described by which the nucleolus organizer regions of mammalian chromosomes, including those of mouse chromosomes, are stained selectively.     

The karyotype,C-bands and AgNO3-bands of a lungless salamander from Korea:Onychodactylus fischeri (Boulenger) (Amphibia,Urodela)

The karyotype of a lungless salamander,Onychodactylus fischeri, from Korea was analyzed and compared with that of the Japanese congeneric species,O. japonicus. In both species the diploid karyotype consists of78 chromosomes, including 6 pairs of large chromosomes, 6 pairs of medium-sized ones, and the remaining 27 pairs of microchromosomes. The chromosome number ofO. fischeri, 2n=78, is, like that ofO. japonicus, the largest so far reported in the order Urodela. C-banding showed that constitutive heterochromatin inO. fischeri was mainly in the centromeric regions and near the secondary constrictions of the large chromosomes. AgNO₃-bands were located in the secondary constrictions associated with C-band heterochromatin.     

A simple one-step procedure for staining the nucleolus organizer regions

Summary A simple silver staining technique for routine use is described by which the nucleolus organizer regions of mammalian chromosomes, including those of mouse chromosomes, are stained selectively.Acknowledgment. We would like to thank Prof. Dr K. Sperling for critical and helpful discussion.     

Cytogenetic evidence for hemizygosity at the thymidine kinase locus in P388 mouse lymphoma cells

A detailed cytogenetic investigation was carried out on P388 mouse lymphoma cells. The cells have a mean chromosome number of 36.86 with a mode and median of 37 chromosomes. G-banding analysis of 12 spreads revealed a total of 15 marker chromosomes with chromosome 11, the determinant of thymidine kinase, being present only in single copy per cell. It is therefore concluded that the P388 cell line is hemizygous at the thymidine kinase locus. Thymidine kinase activities were assayed in P388 cells and two other malignant cell lines, clone 707 Friend mouse leukaemia cells and L5178Y mouse lymphoma cells. No clear relationship was observed between enzyme activity and gene dosage.     

Ultrastructural identification of the nucleolus organizer by the silver staining technic]

The nucleolus organizer regions can be selectively stained in metaphase chromosome preparations by the Goodpasture and Bloom's technique which was adapted to electron microscopy analysis of cells during interphase. Using this technique, a selective accumulation of silver grains was observed over nucleolus light areas. This selective accumulation allows the identification of the interphase fibrillar centers as the nucleolus organizer regions. Ultrastructural relationships between fibrillar centers and dense fibrillar component are discussed.     

Heterochromatin associated with active versus inactive centromeres of mouse replicates at different times

Summary A subline of mouse L-cells carries a dicentric chromosome in which one centromere always separates prematurely. This centromere is not involved in the dynamics of chromosome migration and is considered inactive. By use of anti-BRdU antibody binding to BRdU-treated chromosomes it is shown that the pericentric constitutive heterochromatin associated with the prematurely separating centromere replicates earlier than its counterpart associated with the active centromere and even before several euchromatic regions in the genome. These results point to a possible mechanism by which dicentric chromosomes segregate equationally.     

Predominant regulators of tubulin monomer–polymer partitioning and their implication for cell polarization

The microtubule-system organizes the cytoplasm during interphase and segregates condensed chromosomes during mitosis. Four unrelated conserved proteins, XMAP215/Dis1/TOGp, MCAK, MAP4 and Op18/stathmin, have all been implicated as predominant regulators of tubulin monomer–polymer partitioning in animal cells. However, while studies employing the Xenopus egg extract model system indicate that the partitioning is largely governed by the counteractive activities of XMAP215 and MCAK, studies of human cell lines indicate that MAP4 and Op18 are the predominant regulators of the interphase microtubule-array. Here, we review functional interplay of these proteins during interphase and mitosis in various cell model systems. We also review the evidence that MAP4 and Op18 have interphase-specific, counteractive and phosphorylation-inactivated activities that govern tubulin subunit partitioning in many mammalian cell types. Finally, we discuss evidence indicating that partitioning regulation by MAP4 and Op18 may be of significance to establish cell polarity.     

How to halve ploidy: lessons from budding yeast meiosis

Maintenance of ploidy in sexually reproducing organisms requires a specialized form of cell division called meiosis that generates genetically diverse haploid gametes from diploid germ cells. Meiotic cells halve their ploidy by undergoing two rounds of nuclear division (meiosis I and II) after a single round of DNA replication. Research in Saccharomyces cerevisiae (budding yeast) has shown that four major deviations from the mitotic cell cycle during meiosis are essential for halving ploidy. The deviations are (1) formation of a link between homologous chromosomes by crossover, (2) monopolar attachment of sister kinetochores during meiosis I, (3) protection of centromeric cohesion during meiosis I, and (4) suppression of DNA replication following exit from meiosis I. In this review we present the current understanding of the above four processes in budding yeast and examine the possible conservation of molecular mechanisms from yeast to humans.     

Putting CENP-A in its place

The centromere is the chromosomal region that directs kinetochore assembly during mitosis in order to facilitate the faithful segregation of sister chromatids. The location of the human centromere is epigenetically specified. The presence of nucleosomes that contain the histone H3 variant, CENP-A, are thought to be the epigenetic mark that indicates active centromeres. Maintenance of centromeric identity requires the deposition of new CENP-A nucleosomes with each cell cycle. During S-phase, existing CENP-A nucleosomes are divided among the daughter chromosomes, while new CENP-A nucleosomes are deposited during early G1. The specific assembly of CENP-A nucleosomes at centromeres requires the Mis18 complex, which recruits the CENP-A assembly factor, HJURP. We will review the unique features of centromeric chromatin as well as the mechanism of CENP-A nucleosome deposition. We will also highlight a few recent discoveries that begin to elucidate the factors that temporally and spatially control CENP-A deposition.     

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.     

Visualization of centromeric spots in the chromosomes by UV-light exposure

Summary When exposed to UV-light in the presence of Hoechst 33258, chromosomes ofAllium cepa were progressively photolyzed with increasing length of exposure; they retained their delineated contours, centromeric spots and sometimes secondary constriction bands.     

Giemsa centromere staining ofHumulus lupulus L. Metaphase chromosomes

Summary Giemsa staining of hop (Humulus lupulus L.) chromosomes at metaphase revealed kinetochore-like structures in the centromeric region.The authors are grateful to Dr C. K. Atal, for his keen interest and constant encouragement in the present study.     

High resolution of heterochromatin of Drosophila melanogaster by distamycin A

A DNA-binding AT-specific antibiotic, distamycin A, was used as inhibitor of the condensation process of the heterochromatic regions in Drosophila melanogaster embryonic cells. By this treatment the structural organization of heterochromatin at interphase is preserved until metaphase. The different patterns observed are interpreted as chronological steps in the condensation process.     

Genotoxic effects of dithane M-45 on the bone marrow cells of mice in vivo

Summary Genotoxic effects of dithane M-45 were studied on the bone marrow cells of male albino mice (Lacca strain) in vivo. Different doses (30 mg, 40 mg and 300 mg/kg b.wt) of dithane M-45 were injected intraperitoneally and their effects were investigated after time intervals of 1, 2, 5 and 10 days. The chromosomal aberrations observed in the bone marrow cells of male mice after treatment with dithane M-45 were fragments, rings, dicentric chromosomes, terminal chromatid deletions, chromatid gaps and breaks. In addition to these chromosomal aberrations, physiological effects such as uneven stretching of chromatin material, end-to-end chromosomal associations, exchange configurations, clumping, stickiness and centromeric associations were also observed.     

Genotoxic effects of dithane M-45 on the bone marrow cells of mice in vivo

Genotoxic effects of dithane M-45 were studied on the bone marrow cells of male albino mice (Lacca strain) in vivo. Different doses (30 mg, 40 mg and 300 mg/kg b.wt) of dithane M-45 were injected intraperitoneally and their effects were investigated after time intervals of 1, 2, 5 and 10 days. The chromosomal aberrations observed in the bone marrow cells of male mice after treatment with dithane M-45 were fragments, rings, dicentric chromosomes, terminal chromatid deletions, chromatid gaps and breaks. In addition to these chromosomal aberrations, physiological effects such as uneven stretching of chromatin material, end-to-end chromosomal associations, exchange configurations, clumping, stickiness and centromeric associations were also observed.     

Étude cytologique d'une translocation chromosomeY-autosome chez la souris

Summary A male mouse with one translocation involving theY-chromosome has been observed in the F1 offspring of a male mouse given 300 R of X-irradiation. 6 types of spermatocytes were recorded. Our observations appear to confirm that the acrocentricX of the mouse associates end-to-end with theY at its centromeric end.