Chorionic villus metaphase chromosomes and interphase nuclei analysed by chromosomalin situ suppression (CISS) hybridization

Hooded rats were given an intraperitoneal injection of 3H-tyrosine, and killed in pairs 10 min, 30 min, 12 h, 36 h, 7 days, and 30 days later. A piece of skin with white growing hair, and the tongue, were taken from each animal and radioautographs were prepared. Silver grains were counted over whole nuclei and whole mitotic figures of the germinal cells and whole nuclei of differentiating cells of both tissues. It was found that the interphase nuclei have significantly more silver grains over them than the chromosomes at all stages of mitosis and there are virtually no grains over metaphase, anaphase, and early telophase chromosomes in both tissues of all the animals killed up to 36 h after the injection. The difference between the grain counts over the interphase nuclei and the chromosomes of dividing cells is at least 20-fold at 30 min in the hair matrix, at least 5-fold at 30 min in the tongue and at 36 h in both tissues. It was established that the differences observed between the radioactivities of the nuclei and chromosomes of mitotic figures are real from estimates of: the radioactivity of the cell cytoplasm, volumes of the metaphase chromosomes and interphase nuclei within 1µ of the photographic emulsion, and the volumes of cytoplasm separating the photographic emulsion and these structures. No protein synthesis was demonstrable in the chromosomes during metaphase, anaphase, and early telophase. Nuclear proteins leave the chromosomes during prophase and prometaphase and return to the nucleus during late telophase. The cells in the matrix and upper bulb of the growing hair follicle and those in the germinal, prickle, and granular cell layers of the tongue are in different functional states; 30 min after injection of 3H-tyrosine they have different amounts of it in their nuclear proteins. It is suggested that the amount incorporated into each nucleus is related to the rate at which proteins are being synthesized by the cell.

Download Full-text

A Light Microscope Study of Linker Histone Distribution in Rat Metaphase Chromosomes and Interphase Nuclei

Experimental Cell Research ◽

10.1006/excr.1993.1115 ◽

1993 ◽

Vol 206 (1) ◽

pp. 16-26 ◽

Cited By ~ 25

Author(s):

John W. Breneman ◽

Peter Yau ◽

Raymond L. Teplitz ◽

E.Morton Bradbury

Keyword(s):

Microscope Study ◽

Light Microscope ◽

Linker Histone ◽

Metaphase Chromosomes ◽

Interphase Nuclei

Download Full-text

Characterization of telomeric repeats in metaphase chromosomes and interphase nuclei of Syrian Hamster Fibroblasts

Molecular Cytogenetics ◽

10.1186/1755-8166-5-37 ◽

2012 ◽

Vol 5 (1) ◽

pp. 37 ◽

Cited By ~ 8

Author(s):

Liudmila V Solovjeva ◽

Sergey Demin ◽

Nadezhda M Pleskach ◽

Maria O Kuznetsova ◽

Maria P Svetlova

Keyword(s):

Syrian Hamster ◽

Metaphase Chromosomes ◽

Interphase Nuclei ◽

Telomeric Repeats

Download Full-text

PROPHASING OF INTERPHASE NUCLEI AND INDUCTION OF NUCLEAR ENVELOPES AROUND METAPHASE CHROMOSOMES IN HELA AND CHINESE HAMSTER HOMO- AND HETEROKARYONS

The Journal of Cell Biology ◽

10.1083/jcb.62.1.104 ◽

1974 ◽

Vol 62 (1) ◽

pp. 104-113 ◽

Cited By ~ 22

Author(s):

Yoshitaka Obara ◽

Lee S. Chai ◽

Herbert Weinfeld ◽

Avery A. Sandberg

Keyword(s):

Nuclear Envelope ◽

Hela Cells ◽

Interphase Nucleus ◽

Chinese Hamster ◽

Binucleate Cell ◽

Metaphase Chromosomes ◽

Interphase Nuclei ◽

Multinucleate Cells ◽

Interphase Cells ◽

Nuclear Envelope Formation

Fusing human HeLa metaphase cells with HeLa interphase cells resulted within 30 min in either of two phenomena in the resultant binucleate cell: either prophasing of the interphase nucleus or formation of a normal-appearing nuclear envelope around the metaphase chromosomes. The frequency of either occurrence was strongly dependent on environmental pH. At pH's of 6.6–8.0, prophasing predominated; at pH 8.5 nuclear envelope formation predominated. Additionally, the frequencies of the two events in multinucleate cells depended on the metaphase/interphase ratio. When the ratio was 0.33 nuclear envelope formation predominated; when it was 2.0 prophasing predominated. In their general features, the results with fused HeLa cells resembled those reported earlier with fused Chinese hamster Don cells. However, the results provided an indication that between pH 6.6 and 8.0 the HeLa metaphase cells possessed a much greater capacity than the Don metaphase cells to induce prophasing. Fusion of Don metaphase cells with HeLa interphase cells or of Don interphase cells with HeLa metaphase cells at pH 8.0 resulted in nuclear envelope formation or prophasing in each kind of heterokaryon. As in the homokaryons, the frequencies of the two events in the heterokaryons depended on the metaphase/interphase ratio. The statistics of prophasing and nuclear envelope formation in the homo- and heterokaryon populations were consistent with the notion that disruption or formation of the nuclear envelope depends on the balance attained between disruptive and formative processes.

Download Full-text

INDUCTION OF PROPHASE IN INTERPHASE NUCLEI BY FUSION WITH METAPHASE CELLS

The Journal of Cell Biology ◽

10.1083/jcb.54.1.120 ◽

1972 ◽

Vol 54 (1) ◽

pp. 120-132 ◽

Cited By ~ 42

Author(s):

Sei-Ichi Matsui ◽

Hiroshi Yoshida ◽

Herbert Weinfeld ◽

Avery A. Sandberg

Keyword(s):

Cell Fusion ◽

Sendai Virus ◽

Morphological Changes ◽

Metaphase Chromosomes ◽

Interphase Nuclei ◽

Interphase Cell ◽

Binucleate Cells ◽

Sequence Of Events ◽

Interphase Cells ◽

Nuclear Stage

Fusion of an interphase cell with a metaphase cell results in profound changes in the interphase chromatin that have been called "chromosome pulverization" or "premature chromosome condensation" In addition to the usual light microscopy, the nature of the changes has been investigated in the present study with electron microscopy and biochemical techniques Metaphase and interphase cells were mixed and fused at 37°C by means of ultraviolet-inactivated Sendai virus. After cell fusion, morphological changes in interphase nuclei occurred only in binucleate cells which contained one intact set of metaphase chromosomes Irrespective of the nuclear stage at the time of cell fusion, the morphologic changes that occurred 5–20 min later simulated very closely a sequence of events that characterizes the normal G2-prophase transition. Radioautography revealed that, late in the process, substantial amounts of RNA and probably protein were transferred from the interphase nucleus into the cytoplasm of fused cells. Thus, the findings indicate the existence in metaphase cells of factor(s) which are capable of initiating biochemical and morphological events in interphase nuclei intrinsic to the normal mitotic process.

Download Full-text

High frequency of monoallelic retinoblastoma gene deletion in B-cell chronic lymphoid leukemia shown by interphase cytogenetics

Blood ◽

10.1182/blood.v81.8.2118.bloodjournal8182118 ◽

1993 ◽

Vol 81 (8) ◽

pp. 2118-2124

Author(s):

S Stilgenbauer ◽

H Dohner ◽

M Bulgay-Morschel ◽

S Weitz ◽

M Bentz ◽

...

Keyword(s):

In Situ Hybridization ◽

B Cell ◽

Suppressor Gene ◽

Chromosomal Locus ◽

Metaphase Chromosomes ◽

Interphase Nuclei ◽

Lymphoid Leukemia ◽

Interphase Cytogenetics ◽

Conventional Cytogenetic Analysis

Inactivation of the retinoblastoma tumor-suppressor gene (RB-1) has been associated with tumorigenicity in various human malignancies. In chronic lymphoid leukemias of B-cell origin (B-CLL) an involvement of RB-1 has been suggested based on cytogenetic data. We examined RB-1 and its chromosomal locus 13q14 in 35 cases of B-CLL by dual-color in situ hybridization to interphase nuclei and by G-banding analysis of metaphase chromosomes. In one patient (pt) a monosomy 13, and in three other pts deletions involving or encompassing band 13q14 were detected by conventional cytogenetic analysis. In contrast, in situ hybridization to interphase nuclei showed a monoallelic RB-1 deletion in 11 cases (31%). One pt showed a translocation with the breakpoint in 13q1?4 on G-banding, but on in situ hybridization analysis the RB-1 signals were not affected. Our data show that RB-1 deletions can be diagnosed accurately by in situ hybridization on the one-cell level. The frequency of RB-1 deletions detected in this study is significantly higher than previously assumed in B-CLL, and seems to be in the same range as in retinoblastoma.

Download Full-text

Use of fluorescence in situ hybridization to study the arrangement of DNA sequences in normal and disease-related chromosomes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100140117 ◽

1995 ◽

Vol 53 ◽

pp. 748-749

Author(s):

Barbara J. F. Trask ◽

Hillary Massa ◽

Cynthia Friedman ◽

Richard Esposito ◽

Ger van den Engh ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Dna Sequences ◽

Single Copy ◽

Two Dimensions ◽

Genetic Maps ◽

Epifluorescence Microscopy ◽

Metaphase Chromosomes ◽

Interphase Nuclei

The sites of specific DNA sequences can be fluorescently tagged by fluorescence in situ hybridization (FISH). Different sequences can be labeled with different fluorochromes so that their arrangement can be studied using epifluorescence microscopy. The distances between points on the same or different chromosomes can be determined easily in a large number of interphase nuclei or metaphase chromosomes. A variety of probe types, ranging from single-copy sequences to highly repeated sequences can be employed. Our work has focussed on the analysis of hybridization patterns in two dimensions using conventional fluorescence microscopy.We have used FISH to study various aspects of genome organization that are difficult to study using other techniques. Examples of these applications will be presented.FISH is now the method of choice for determining the chromosomal location of DNA sequences. DNA sequences can be positioned in the genome with <1:1000 accuracy (to a 3-Mbp region within a 3000-Mbp genome). Through FISH, the cytogenetic, physical and genetic maps of chromosomes can be linked.

Download Full-text

The putative nuclear receptor mediator TIF1alpha is tightly associated with euchromatin

Journal of Cell Science ◽

10.1242/jcs.112.11.1671 ◽

1999 ◽

Vol 112 (11) ◽

pp. 1671-1683 ◽

Cited By ~ 2

Author(s):

E. Remboutsika ◽

Y. Lutz ◽

A. Gansmuller ◽

J.L. Vonesch ◽

R. Losson ◽

...

Keyword(s):

Rna Polymerase Ii ◽

Nuclear Receptors ◽

Target Genes ◽

Es Cells ◽

Embryonic Stem ◽

Chromosomal Protein ◽

Metaphase Chromosomes ◽

Interphase Nuclei ◽

Ec Cells ◽

Early Mouse

Ligand-dependent transcriptional regulation by nuclear receptors is believed to be mediated by intermediary factors (TIFs) acting on remodelling of the chromatin structure and/or the activity of the transcriptional machinery. The putative transcriptional mediator TIF1alpha is a nuclear protein kinase that has been identified via its interaction with liganded nuclear receptors, including retinoic acid (RAR), retinoid X (RXR) and estrogen (ER) receptors. Here, we demonstrate that TIF1alpha is a non-histone chromosomal protein tightly associated with highly accessible euchromatic regions of the genome. Immunofluorescence confocal microscopy reveals that TIF1alpha exhibits a finely granular distribution in euchromatin of interphase nuclei, while it is mostly excluded from condensed chromatin and metaphase chromosomes. Immunoelectron microscopy shows that, in contrast to the heterochromatin protein HP1alpha, most of TIF1alpha is associated with euchromatin, where it is preferentially localised on regions known to be sites for RNA polymerase II (perichromatin fibrils and borders between euchromatin and heterochromatin). Early mouse embryos as well as embryonal carcinoma (EC) and embryonic stem (ES) cells express high levels of TIF1alpha. These levels dramatically decrease during organogenesis and upon differentiation of P19 EC cells, indicating that TIF1alpha is preferentially expressed in undifferentiated pluripotent cells in the course of development. Therefore, TIF1alpha could belong to a novel class of chromatin-associated TIFs that facilitate the access of transregulators (e.g. liganded nuclear receptors) to their cognate sites in target genes, thereby participitating in the epigenetic control of transcription during embryonic development and cell differentiation.

Download Full-text

Nucleolus-associated bodies in meristematic cells of two plant species (Cicer arietinum and Leucaena glauca) with different ploidy levels

Canadian Journal of Botany ◽

10.1139/b91-172 ◽

1991 ◽

Vol 69 (6) ◽

pp. 1329-1336 ◽

Cited By ~ 8

Author(s):

J. G. Lafontaine ◽

B. T. Luck ◽

S. Gugg

Keyword(s):

Cicer Arietinum ◽

Plant Species ◽

Diploid Species ◽

Nucleolar Organizer ◽

Ultrastructural Level ◽

Serial Sections ◽

Metaphase Chromosomes ◽

Interphase Nuclei ◽

Ploidy Levels ◽

Meristematic Cells

Light microscopy has shown that plant interphase nuclei contain small, roundish bodies, some of which may be closely associated with the nucleolar surface. Serial sections were used to determine the location, size, and number of these nucleolus-associated bodies in two plant species having different ploidy levels. In Cicer arietinum, a diploid species, one or two such bodies were observed, whereas in Leucaena glauca, an octoploid species, four to six nucleolus-associated bodies were present. At the ultrastructural level, these bodies consistently exhibited a distinct fibrillogranular texture and were located close to segments of the interphase nucleolar organizer track, a meandering, coarse, filamentous structure particularly well developed within many plant species and known to consist partly of chromatin. The fact that the number of these bodies closely matches that of the satellite-bearing metaphase chromosomes suggests that they may represent terminal segments of the nucleolar chromosomes. Other equally plausible interpretations of the nature of the NABs are also considered. Key words: plant interphase nucleus, nucleolus-associated bodies, satellites.

Download Full-text