C-banding plus fluorochrome staining shows differences in C-, G-, and R-bands in human and mouse metaphase chromosomes

Genome ◽  
1995 ◽  
Vol 38 (5) ◽  
pp. 864-868 ◽  
Author(s):  
J. L. Bella ◽  
J. Gosálvez ◽  
J. L. Fernández
Keyword(s):  
Mouse Cell ◽  
Structural Factors ◽  
Metaphase Chromosomes ◽  
Fluorochrome Staining ◽  
Banding Patterns ◽  
C Banding ◽  
Dapi Banding ◽  
Species Specific ◽  
L929 Mouse ◽  
Human And Mouse

C-banded slides stained with DAPI or chromomycin A3 show different banding patterns between human and L929 mouse cell line metaphase chromosomes, which are also different from those obtained with standard Giemsa C-banding or fluorochrome staining. Human metaphase chromosomes pretreated for C-banding and stained with DAPI show simultaneous C– and DA–DAPI banding patterns, whilst the mouse metaphase chromosomes show both C-banding and G/Q banding like patterns. However, the chromomycin A3 staining of pre-C-banded metaphase chromosomes reveals conspicuous R-banding in man that is absent in mouse. Chromatin species-specific structural factors would explain these results, which prevent simple comparisons of R-, G-, and C-bands among different organisms. The markers induced by this technique may be of practical use for chromosome identification in human–mouse somatic cell hybridization cultures.Key words: mammalian cytogenetics, chromosome structure, chromosome banding, fluorochrome staining.

Chromosome Analysis of Two Related Heteroploid Mouse Cell Lines by Quinacrine Fluorescence

1973 ◽  
Vol 12 (1) ◽  
pp. 263-274
Author(s):  
P. W. ALLDERDICE ◽  
O. J. MILLER ◽  
D. A. MILLER ◽  
D. WARBURTON ◽  
P. L. PEARSON ◽  
...  
Keyword(s):  
Cell Lines ◽  
Chromosome Analysis ◽  
Mouse Cell ◽  
Previous Method ◽  
Structural Rearrangements ◽  
Metaphase Chromosomes ◽  
Detailed Characterization ◽  
Banding Patterns ◽  
Fluorescent Banding ◽  
Quinacrine Fluorescence

The fluorescent banding patterns of quinacrine-stained metaphase chromosomes have been studied in 2 related mouse cell lines, A9 and a malignant derivative of A9, A9HT. In both cell lines virtually every chromosome has a distinctive banding pattern which permits its recognition. More than three quarters of the chromosomes have structural rearrangements, but the origin of nearly two thirds of the chromosomes could be determined by their banding patterns. The quinacrine fluorescence technique permits far more detailed characterization and comparison of heteroploid cell lines than any previous method. A9 and A9HT are karyologically quite similar, with many of the same marker chromosomes. There are, however, characteristic differences. A9HT, although it has a smaller average number of chromosomes per cell, appears to be more heterogeneous.

scholarly journals Conserved cell types with divergent features between human and mouse cortex

2018 ◽  
Author(s):  
Rebecca D Hodge ◽  
Trygve E Bakken ◽  
Jeremy A Miller ◽  
Kimberly A Smith ◽  
Eliza R Barkan ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Cognitive Abilities ◽  
Cell Types ◽  
Mouse Cell ◽  
Middle Temporal Gyrus ◽  
Cellular Architecture ◽  
Mouse Cortex ◽  
Single Nucleus ◽  
Species Specific ◽  
Human And Mouse

AbstractElucidating the cellular architecture of the human neocortex is central to understanding our cognitive abilities and susceptibility to disease. Here we applied single nucleus RNA-sequencing to perform a comprehensive analysis of cell types in the middle temporal gyrus of human cerebral cortex. We identify a highly diverse set of excitatory and inhibitory neuronal types that are mostly sparse, with excitatory types being less layer-restricted than expected. Comparison to a similar mouse cortex single cell RNA-sequencing dataset revealed a surprisingly well-conserved cellular architecture that enables matching of homologous types and predictions of human cell type properties. Despite this general conservation, we also find extensive differences between homologous human and mouse cell types, including dramatic alterations in proportions, laminar distributions, gene expression, and morphology. These species-specific features emphasize the importance of directly studying human brain.

scholarly journals Comparison of reprogramming factor targets reveals both species-specific and conserved mechanisms in early iPS cells

2017 ◽  
Author(s):  
Kai Fu ◽  
Constantinos Chronis ◽  
Abdenour Soufi ◽  
Giancarlo Bonora ◽  
Miguel Edwards ◽  
...  
Keyword(s):  
Regulatory Networks ◽  
Target Genes ◽  
Human Fibroblasts ◽  
Mouse Cell ◽  
Binding Motifs ◽  
Reprogramming Factors ◽  
Early Mouse ◽  
Species Specific ◽  
Human And Mouse ◽  
Syntenic Regions

AbstractBoth human and mouse fibroblasts can be reprogrammed to pluripotency with Oct4, Sox2, Klf4, and c-Myc (OSKM) transcription factors. While both systems generate pluripotency, human reprogramming takes considerably longer than mouse. To assess additional similarities and differences, we sought to compare the binding of the reprogramming factors between the two systems. In human fibroblasts, the OSK factors initially target many more closed chromatin sites compared to mouse. Despite this difference, the intra- and intergenic distribution of target sites, target genes, primary binding motifs, and combinatorial binding patterns between the reprogramming factors are largely shared. However, while many OSKM binding events in early mouse cell reprogramming occur in syntenic regions, only a limited number is conserved in human. In summary, these findings suggest similar general effects of OSKM binding across these two species, even though the detailed regulatory networks have diverged significantly.

5-Methylcytosine-Rich Heterochromatin in Reptiles

2019 ◽  
Vol 157 (1-2) ◽  
pp. 53-64 ◽  
Author(s):  
Michael Schmid ◽  
Claus Steinlein ◽  
Alina M. Reiter ◽  
Michail Rovatsos ◽  
Marie Altmanová ◽  
...  
Keyword(s):  
Indirect Immunofluorescence ◽  
Sex Chromosomes ◽  
Dna Sequences ◽  
Experimental Approach ◽  
Constitutive Heterochromatin ◽  
Banding Patterns ◽  
Turtle Species ◽  
C Banding ◽  
Species Specific ◽  
Chromosome Regions

An experimental approach using monoclonal anti-5-methylcytosine antibodies and indirect immunofluorescence was elaborated for detecting 5-methylcytosine-rich chromosome regions in reptilian chromosomes. This technique was applied to conventionally prepared mitotic metaphases of 2 turtle species and 12 squamate species from 8 families. The hypermethylation patterns were compared with C-banding patterns obtained by conventional banding techniques. The hypermethylated DNA sequences are species-specific and are located in constitutive heterochromatin. They are highly reproducible and often found in centromeric, pericentromeric, and interstitial positions of the chromosomes. Heterochromatic regions in differentiated sex chromosomes are particularly hypermethylated.

scholarly journals Authentication, characterization and contamination detection of cell lines, xenografts and organoids by barcode deep NGS sequencing

2020 ◽  
Vol 2 (3) ◽  
Author(s):  
Xiaobo Chen ◽  
Wubin Qian ◽  
Zhenzhen Song ◽  
Qi-Xiang Li ◽  
Sheng Guo
Keyword(s):  
Cell Lines ◽  
Low Cost ◽  
Human Cell Line ◽  
Mouse Cell ◽  
Single Nucleotide ◽  
And Gender ◽  
Species Specific ◽  
Human And Mouse ◽  
Higher Sensitivity ◽  
Short Tandem

Abstract Misidentification and contamination of biobank samples (e.g. cell lines) have plagued biomedical research. Short tandem repeat (STR) and single-nucleotide polymorphism assays are widely used to authenticate biosamples and detect contamination, but with insufficient sensitivity at 5–10% and 3–5%, respectively. Here, we describe a deep NGS-based method with significantly higher sensitivity (≤1%). It can be used to authenticate human and mouse cell lines, xenografts and organoids. It can also reliably identify and quantify contamination of human cell line samples, contaminated with only small amount of other cell samples; detect and quantify species-specific components in human–mouse mixed samples (e.g. xenografts) with 0.1% sensitivity; detect mycoplasma contamination; and infer population structure and gender of human samples. By adopting DNA barcoding technology, we are able to profile 100–200 samples in a single run at per-sample cost comparable to conventional STR assays, providing a truly high-throughput and low-cost assay for building and maintaining high-quality biobanks.

Characterization of Hordeum chilense chromosomes by C-banding and in situ hybridization using highly repeated DNA probes

Genome ◽  
1995 ◽  
Vol 38 (3) ◽  
pp. 435-442 ◽  
Author(s):  
A. Cabrera ◽  
B. Friebe ◽  
J. Jiang ◽  
B. S. Gill
Keyword(s):  
In Situ Hybridization ◽  
Relative Length ◽  
Hordeum Chilense ◽  
Addition Lines ◽  
Metaphase Chromosomes ◽  
Banding Patterns ◽  
Rdna Probe ◽  
26S Rdna ◽  
C Banding

C-banding patterns of Hordeum chilense and of Triticum aestivum 'Chinese Spring' – H. chilense disomic addition lines were analyzed and compared with in situ hybridization patterns using a biotin-labeled highly repetitive Triticum tauschii DNA sequence, pAs1, and a wheat 18S–26S rDNA probe. All seven H. chilense chromosomes pairs and the added H. chilense chromosomes present in the addition lines were identified by their characteristic C-banding pattern. Chromosome morphology and banding patterns were similar to those of the corresponding chromosomes present in the parent H. chilense accession. A C-banded karyotype of the added H. chilense chromosomes was constructed and chromosome lengths, arm ratios, and relative length, as compared with chromosome 3B, were determined. The probe pAs1 was found to hybridize to specific areas on telomeres and interstitial sites along the chromosomes, allowing the identification of all seven pairs of the H. chilense chromosomes. Comparison of the patterns of distribution of the hybridization sites of clone pAs1 in the T. tauschii and H. chilense chromosomes was carried out by in situ hybridization on somatic metaphase chromosomes of the HchHchDD amphiploid. In situ hybridization using the 18S–26S rDNA probe confirmed that the H. chilense chromosomes 5Hch and 6Hch were carrying nucleolus organizer regions. The results are discussed on the basis of phylogenetic relationships between D and Hch genomes.Key words: Hordeum, Triticum, C-banding, in situ hybridization, phylogeny.

scholarly journals Heterochromatin analysis in the fish species Liposarcus anisitsi (siluriformes) and Leporinus elongatus (characiformes)

1999 ◽  
Vol 22 (1) ◽  
pp. 39-44 ◽  
Author(s):  
Roberto Ferreira Artoni ◽  
Wagner Franco Molina ◽  
Luis Antonio Carlos Bertollo ◽  
Pedro Manoel Galetti Junior
Keyword(s):  
Thermal Stability ◽  
Freshwater Fish ◽  
Fish Species ◽  
Saline Solution ◽  
Fluorochrome Staining ◽  
Banding Patterns ◽  
C Banding ◽  
Heterochromatin Differentiation ◽  
First Time ◽  
Thermal Stability Of

The chromosomes of two neotropical freshwater fish species, namely Liposarcus anisitsi (Siluriformes, Loricariidae) and Leporinus elongatus (Characiformes, Anostomidae), were investigated by means of C-banding, Ag-NORs, fluorochrome staining and banding by hot saline solution (HSS) treatment, to reveal patterns of heterochromatin differentiation. The karyotype of L. anisitsi is described for the first time. Staining with the GC-specific fluorescent antibiotic mithramycin (MM) revealed bright signals in some C-banded blocks in both species, suggesting that these MM+ heterochromatin contains GC-rich DNA. Banding by denaturation employing HSS, followed by Giemsa staining, yielded corresponding results documenting the thermal stability of GC-rich DNA part of heterochromatin positive after C-banding. In L. elongatus the Ag-NOR also followed the above banding patterns. However, in L. anisitsi the Ag-NOR was MM+ but negatively stained after C-banding and HSS treatment. L. elongatus also showed C-banded segments that were negative for mithramycin staining and HSS treatment. The results obtained evidence the heterochromatin heterogeneity in these fish species.

Some Structural Features of Metaphase Chromosomes of Mice and Men

1967 ◽  
Vol 25 ◽  
pp. 190-191
Author(s):  
Godfrey C. Hoskins ◽  
Betty B. Hoskins
Keyword(s):  
Electron Microscopy ◽  
Electron Micrographs ◽  
Structural Features ◽  
Metaphase Chromosomes ◽  
The Future ◽  
Of Mice And Men ◽  
Mouse Cells ◽  
Human And Mouse

Metaphase chromosomes from human and mouse cells in vitro are isolated by micrurgy, fixed, and placed on grids for electron microscopy. Interpretations of electron micrographs by current methods indicate the following structural features.Chromosomal spindle fibrils about 200Å thick form fascicles about 600Å thick, wrapped by dense spiraling fibrils (DSF) less than 100Å thick as they near the kinomere. Such a fascicle joins the future daughter kinomere of each metaphase chromatid with those of adjacent non-homologous chromatids to either side. Thus, four fascicles (SF, 1-4) attach to each metaphase kinomere (K). It is thought that fascicles extend from the kinomere poleward, fray out to let chromosomal fibrils act as traction fibrils against polar fibrils, then regroup to join the adjacent kinomere.

Advances in imaging SIMS studies of stained and BrdU-labelled human metaphase chromosomes

1995 ◽  
Vol 53 ◽  
pp. 932-933
Author(s):  
R. Levi-Setti ◽  
J. M. Chabala ◽  
R. Espinosa ◽  
M. M. Le Beau
Keyword(s):  
High Performance ◽  
Secondary Ion Mass Spectrometry ◽  
Analytical Sensitivity ◽  
Metaphase Chromosomes ◽  
Banding Patterns ◽  
Sector Mass Spectrometer ◽  
Staining Methods ◽  
The University ◽  
Secondary Ion ◽  
Better Than

We have shown previously that isotope-labelled nucleotides in human metaphase chromosomes can be detected and mapped by imaging secondary ion mass spectrometry (SIMS), using the University of Chicago high resolution scanning ion microprobe (UC SIM). These early studies, conducted with BrdU- and 14C-thymidine-labelled chromosomes via detection of the Br and 28CN- (14C14N-> labelcarrying signals, provided some evidence for the condensation of the label into banding patterns along the chromatids (SIMS bands) reminiscent of the well known Q- and G-bands obtained by conventional staining methods for optical microscopy. The potential of this technique has been greatly enhanced by the recent upgrade of the UC SIM, now coupled to a high performance magnetic sector mass spectrometer in lieu of the previous RF quadrupole mass filter. The high transmission of the new spectrometer improves the SIMS analytical sensitivity of the microprobe better than a hundredfold, overcoming most of the previous imaging limitations resulting from low count statistics.

Rye C-banding patterns and meiotic stability of hexaploid triticale (× Triticosecale) selections differing in kernel shriveling

Genome ◽  
1990 ◽  
Vol 33 (5) ◽  
pp. 686-689 ◽  
Author(s):  
Charles M. Papa ◽  
R. Morris ◽  
J. W. Schmidt
Keyword(s):  
Secale Cereale ◽  
Chromosome Banding ◽  
Agronomic Performance ◽  
Hexaploid Triticale ◽  
Kernel Development ◽  
Banding Patterns ◽  
C Banding ◽  
Meiotic Stability ◽  
Metaphase I

Two winter hexaploid triticale populations derived from the same cross were selected on the basis of grain appearance and agronomic performance. The five lines from 84LT402 showed more kernel shriveling than the four lines from 84LT401. The derived lines were analyzed for aneuploid frequencies, rye chromosome banding patterns, and meiotic stability to detect associations with kernel development. The aneuploid frequencies were 16% in 84LT401 and 18% in 84LT402. C-banding showed that both selection groups had all the rye chromosomes except 2R. The two groups had similar telomeric patterns but differed in the long-arm interstitial patterns of 4R and 5R. Compared with lines from 84LT402, those from 84LT401 had significantly fewer univalents and rod bivalents, and more paired arms at metaphase I; fewer laggards and bridges at anaphase I; and a higher frequency of normal tetrads. There were no significant differences among lines within each group for any meiotic character. Since there were no differences within or between groups in telomeric banding patterns, the differences in kernel shriveling and meiotic stability might be due to genotypic factors and (or) differences in the interstitial patterns of 4R and 5R. By selecting plump grains, lines with improved kernel characteristics along with improved meiotic stability are obtainable.Key words: triticale, meiotic stability, C-banding, Secale cereale, heterochromatin.

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