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PubMed1993

Karyotypic analysis of CCRF-CEM and drug-resistant cell lines with stable and unstable ploidy.

Karyotypic studies were performed on methotrexate (MTX)- and vincristine (VCR)-resistant cell lines derived from the human T-cell leukemia CCRF-CEM cell line. We noted karyotypic selection and additional chromosome change associated with acquisition of drug-resistance. Furthermore, we observed that both the parental and MTX-resistant sublines, CEM/MTX R1-3, had a tendency to ploidy change. Karyotypic studies of malignant cells have shown that polyploidy is frequently a consequence of a single sporadic event, followed by growth and selection of the polyploid clone [1]. In this study, however, various karyotypic clones were identified with near- and pseudotetraploid karyotypes that appeared to be derived from different near- and pseudodiploid sidelines. Polyploidy was invariably associated with loss of at least two of the four chromosomes 8 whether the pseudodiploid sideline from which it was derived carried one or both chromosomes 8. In contrast, neither polyploid clones nor loss of chromosome 8 was noted in the CEM/VCR R cells.

Chromosome Aberrations↗
PubMedDate not supplied

Molecular karyotype analysis and mapping of housekeeping genes to chromosomes of selected species complexes of Leishmania.

The molecular karyotypes for 20 reference strains of species complexes of Leishmania were determined by contour-clamped homogeneous electric field (CHEF) electrophoresis. Determination of number/position of chromosome-sized bands and chromosomal DNA locations of housekeeping genes were the two criteria used for differentiating and classifying the Leishmania species. We have established two gel running conditions for optimal separation of chromosomes, which resolved DNA molecules as large as 2,500 kilobase pairs (kb). Chromosomes were polymorphic in number (22-30) and size (200-2,500 kb) of bands among members of five complexes of Leishmania. Although each stock had a distinct karyotype, in general the differences found between strains and/or species within each complex were not clear enough for parasite identification. However, each group showed a specific number of size-concordant DNA molecules, which allowed distinction among the Leishmania complex parasites. Clear differences between the Old and New world groups of parasites or among some New World Leishmania species were also apparent in relation to the chromosome locations of beta-tubulin genes. Based on these results as well as data from other published studies the potential of using DNA karyotype for identifying and classifying leishmanial field isolates is discussed.

Animals↗
PubMed2001

Karyotype analysis of Lilium longiflorum and Lilium rubellum by chromosome banding and fluorescence in situ hybridisation.

Detailed karyotypes of Lilium longiflorum and L. rubellum were constructed on the basis of chromosome arm lengths, C-banding, AgNO3 staining, and PI-DAPI banding, together with fluorescence in situ hybridisation (FISH) with the 5S and 45S rDNA sequences as probes. The C-banding patterns that were obtained with the standard BSG technique revealed only few minor bands on heterologous positions of the L. longiflorum and L. rubellum chromosomes. FISH of the 5S and 45S rDNA probes on L. longiflorum metaphase complements showed overlapping signals at proximal positions of the short arms of chromosomes 4 and 7, a single 5S rDNA signal on the secondary constriction of chromosome 3, and one 45S rDNA signal adjacent to the 5S rDNA signal on the subdistal part of the long arm of chromosome 3. In L. rubellum, we observed co-localisation of the 5S and 45S rDNA sequences on the short arm of chromosomes 2 and 4 and on the long arms of chromosomes 2 and 3, and two adjacent bands on chromosome 12. Silver staining (Ag-NOR) of the nucleoli and NORs in L. longiflorum and L. rubellum yielded a highly variable number of signals in interphase nuclei and only a few faint silver deposits on the NORs of mitotic metaphase chromosomes. In preparations stained with PI and DAPI, we observed both red- and blue-fluorescing bands at different positions on the L. longiflorum and L. rubellum chromosomes. The red-fluorescing or so-called reverse PI-DAPI bands always coincided with rDNA sites, whereas the blue-fluorescing DAPI bands corresponded to C-bands. Based on these techniques, we could identify most of chromosomes of the L. longiflorum and L. rubellum karyotypes.

Chromosome Banding↗
PubMed1993

Preparation and karyotype analysis of mitotic chromosomes of the freshwater sponge Spongilla lacustris.

The present study documents for the first time the karyotype and mitotic chromosomes of a sponge. For the studies the freshwater sponge Spongilla lacustris (Lin. 1758) was used. Its karyotype comprises nine different chromosome pairs ranging in size from 2.1 to < or = 0.7 microns. Changes in size and shape of the chromosomes during the progression of mitosis are documented both light and electron microscopically. The data reveal that the lowest multicellular eukaryotes, the sponges, have already reached a high level of evolution of the mitotic mechanism.

Animals↗
PubMed2006

Karyotype analysis of Gossypium arboreum x G. bickii by genome in situ hybridization.

By using genome in situ hybridization (GISH) on root somatic chromosomes of allotetraploid derived from the cross Gossypium arboreum x G. bickii with genomic DNA (gDNA) of G. bickii as a probe, two sets of chromosomes, consisting of 26 chromosomes each, were easily distinguished from each other by their distinctive hybridization signals. GISH analysis directly proved that the hybrid G. arboreum x G. bickii is an allotetraploid amphiploid. The karyotype formula of the species was 2n = 4x = 52 = 46m (4sat) + 6sm (4sat). We identified four pairs of satellites with two pairs in each sub-genome. FISH analysis using 45S rDNA as a probe showed that the cross G. arboreum x G. bickii contained 14 NORs. At least five pairs of chromosomes in the G sub-genome showed double hybridization (red and blue) in their long arms, which indicates that chromatin introgression from the A sub-genome had occurred.

Chromatin↗
PubMed1998

Cross-species colour segmenting: a novel tool in human karyotype analysis.

We used fluorescence in situ hybridization (FISH) with DNA probes derived from bivariate fluorescence activated flow sorting of primate chromosomes. In cases where human and primate karyotypes differ by chromosome rearrangements, reverse painting of primate probes resulted in a subregional delineation of the human homologous chromosomes. Probes were used from two gibbon species (Hylobates concolor and H. syndactylus) which both showed highly rearranged karyotypes. Hybridization of human chromosomes with painting probes derived from both gibbons showed that, with the exception of human chromosomes 15, 18, 21, 22 and the sex chromosomes, each chromosome was differentiated in at least two and up to six segments. These probes have been used in the analysis of various cases of constitutional chromosomal rearrangements in human pathology including complex intrachromosomal rearrangements. They were also used in a multi colour format (colour segmenting) to differentiate the entire human karyotype into 81 homologous coloured segments with probes derived from H. concolor, and 74 segments with probes derived from H. syndactylus. The addition of colours not only simplifies chromosome identification compared to the analysis of classical banding based on grey values, but colour segmenting also provides simple coloured landmarks for further fine analysis by classical banding.

Animals↗
PubMed2001

Karyotype analysis of tumorigenic human bronchial epithelial cells transformed by chrysolite asbestos using chemically induced premature chromosome condensation technique.

We examined karyotypic changes of tumorigenic human bronchial epithelial cell lines transformed by asbestos fibers. Using Calyculin A mediated premature chromosome condensation (PCC) assay and Giemsa-trypsin banding, we showed that the common changes of all tumorigenic cell lines were the loss of one or two copies of chromosome 5, the monosomy of chromosome 19 and the increased trisomy of chromosome 8. The results indicate that the karyotypic change of chromosome 5, 8 and 19 could play an important role in asbestos-induced tumorigenic conversion of human bronchial epithelial cells from an immortalized to tumorigenic state.

Animals↗
PubMed2005

[Karyotype analysis of Lonicera japonica and L. maackii].

OBJECTIVE: To study Lonicera japonica Thunb. and Lonicera maackii (Rupr. ) Maxim. chromosome number, karyotype, volume. METHODS: Sections combined with micrograph were used to analyse chromosome. RESULTS: Lonicera japonica Thunb. normal diploid 2n = 18; the relative length was 2n = 18 = 2L +8M, + 6M1 + 2S; karyotpye formula was K(2n) = 2X = 18 = 10 sm + 6 m + 2st, which belong to "2A" type; the AS. K% was 66.65%. Lonicera maackii (Rupr.) Maxim. normal diploid 2n = 18; the relative length was 2n = 18 = 4L + 4M, + 10M1; karyotype formula was K(2n) = 2X = 18 = 10 sm + 8m, which belong to"2A" type; the AS. K% was 63.28%. CONCLUSION: The chromosome number, karyotype, and chromosome volume of Lonicera japonica Thunb. and Lonicera maackii (Rupr.) Maxim. were showed clear.

Chromosomes, Plant↗
PubMed1994

Karyotype analysis of the monogenetic trypanosomatid Leptomonas collosoma.

In order to develop a genetic system for the monogenetic trypanosomatids, we have analyzed the molecular karyotype of Leptomonas collosoma based on chromosome separation by clamped homogeneous electric field (CHEF) gel electrophoresis. The chromosome location of 5 RNA coding genes (SL, U6, 5S, 7SL and rRNA) and 2 protein coding genes (for HSP83 and alpha-tubulin) was determined. All of the L. collosoma genes examined were found on at least 2 chromosomes, which differ in size in the range of 100-500 kb, suggesting that the organism is diploid. The weighted sum of L. collosoma chromosomes separated by CHEF analysis was approximately 62 +/- 3 Mb, whereas the genome size determined by FACS was estimated at approx. 80 Mb. This suggests that some of the homologous chromosomes differ in their size. The analysis presented here may facilitate studies on the function of individual genes, and on the genetic stability of this organism.

Animals↗