Effects of 5-azacytidine on the centromeric region of human fibroblasts studied by CREST staining and in situ hybridization on cytokinesis-blocked cells

1996 ◽  
Vol 72 (2-3) ◽  
pp. 219-224 ◽  
Author(s):  
D. Cimini ◽  
C. Tanzarella ◽  
F. Degrassi
Keyword(s):  
In Situ Hybridization ◽  
Centromeric Region ◽  
Human Fibroblasts

scholarly journals Intracellular distribution of histone mRNAs in human fibroblasts studied by in situ hybridization.

1988 ◽  
Vol 85 (2) ◽  
pp. 463-467 ◽  
Author(s):  
J. B. Lawrence ◽  
R. H. Singer ◽  
C. A. Villnave ◽  
J. L. Stein ◽  
G. S. Stein
Keyword(s):  
In Situ Hybridization ◽  
Human Fibroblasts ◽  
Intracellular Distribution ◽  
Histone Mrnas

Characterization of a new 4BS.7HL wheat–barley translocation line using GISH, FISH, and SSR markers and its effect on the β-glucan content of wheat

Genome ◽  
2011 ◽  
Vol 54 (10) ◽  
pp. 795-804 ◽  
Author(s):  
A. Cseh ◽  
K. Kruppa ◽  
I. Molnár ◽  
M. Rakszegi ◽  
J. Doležel ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Ssr Markers ◽  
Repetitive Dna ◽  
Centromeric Region ◽  
Barley Chromosome ◽  
Translocation Chromosome ◽  
Translocation Line ◽  
Special Focus ◽  
Addition Line

A spontaneous interspecific Robertsonian translocation was revealed by genomic in situ hybridization (GISH) in the progenies of a monosomic 7H addition line originating from a new wheat ‘Asakaze komugi’ × barley ‘Manas’ hybrid. Fluorescence in situ hybridization (FISH) with repetitive DNA sequences (Afa family, pSc119.2, and pTa71) allowed identification of all wheat chromosomes, including wheat chromosome arm 4BS involved in the translocation. FISH using barley telomere- and centromere-specific repetitive DNA probes (HvT01 and (AGGGAG)n) confirmed that one of the arms of barley chromosome 7H was involved in the translocation. Simple sequence repeat (SSR) markers specific to the long (L) and short (S) arms of barley chromosome 7H identified the translocated chromosome segment as 7HL. Further analysis of the translocation chromosome clarified the physical position of genetically mapped SSRs within 7H, with a special focus on its centromeric region. The presence of the HvCslF6 gene, responsible for (1,3;1,4)-β-d-glucan production, was revealed in the centromeric region of 7HL. An increased (1,3;1,4)-β-d-glucan level was also detected in the translocation line, demonstrating that the HvCslF6 gene is of potential relevance for the manipulation of wheat (1,3;1,4)-β-d-glucan levels.

scholarly journals Heparin-binding EGF-like growth factor mRNA is upregulated in the peri-infarct region of the remnant kidney model: in vitro evidence suggests a regulatory role in myofibroblast transformation.

1998 ◽  
Vol 9 (8) ◽  
pp. 1464-1473
Author(s):  
G Kirkland ◽  
K Paizis ◽  
L L Wu ◽  
M Katerelos ◽  
D A Power
Keyword(s):  
Endothelial Cells ◽  
In Situ Hybridization ◽  
Growth Factor ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Human Fibroblasts ◽  
Renal Infarction ◽  
Heparin Binding ◽  
Alpha Smooth Muscle Actin

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a potent fibroblast and epithelial cell mitogen that may be important in wound healing. The aim of this study was to determine its distribution and possible function in segmental renal infarction. At day 1 postinfarction, in situ hybridization showed that HB-EGF mRNA was markedly increased by tubular epithelial cells bordering the infarcted zone. At day 3, typical myofibroblasts expressing alpha-smooth muscle actin (alpha-SMA) were present in large numbers at the peri-ischemic border and, over succeeding days, were also seen within the infarcted area. Some of these cells expressed HB-EGF mRNA by in situ hybridization suggesting possible autocrine stimulation. Endothelial cells appeared to be more resistant to ischemia than tubules because some capillaries at the periphery of the infarct, surrounded by infarcted tubules, also expressed HB-EGF mRNA. The staining intensity of HB-EGF mRNA in individual tubules and endothelial cells was maximal at day 5 after infarction, although Northern blots of tissue from the peri-infarct area only showed significantly increased expression of HB-EGF mRNA at days 1 and 3, perhaps reflecting a smaller area of greater intensity of expression at day 5. Because tubular cells expressing high levels of HB-EGF mRNA were directly apposed to myofibroblasts, an attempt was made to determine whether HB-EGF contributed to upregulation of alpha-SMA by human fibroblasts. Although stimulation of the fibroblast cell line MRC-5 with transforming growth factor-beta1 (TGF-beta1) increased alpha-SMA, HB-EGF reduced expression. HB-EGF also strongly inhibited the increased expression of alpha-SMA due to TGF-beta1. Because HB-EGF is a potent fibroblast mitogen and TGF-beta is usually antiproliferative, this study suggests that HB-EGF may contribute to a local balance between fibroblast proliferation and differentiation into myofibroblasts during scarring.

St2-80: a new FISH marker for St genome and genome analysis in Triticeae

Genome ◽  
2017 ◽  
Vol 60 (7) ◽  
pp. 553-563 ◽  
Author(s):  
Long Wang ◽  
Qinghua Shi ◽  
Handong Su ◽  
Yi Wang ◽  
Lina Sha ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genome Analysis ◽  
Centromeric Region ◽  
Repetitive Sequences ◽  
Terminal Region ◽  
Oligonucleotide Primer ◽  
Dot Blot ◽  
D Genome ◽  
Ploidy Levels

The St genome is one of the most fundamental genomes in Triticeae. Repetitive sequences are widely used to distinguish different genomes or species. The primary objectives of this study were to (i) screen a new sequence that could easily distinguish the chromosome of the St genome from those of other genomes by fluorescence in situ hybridization (FISH) and (ii) investigate the genome constitution of some species that remain uncertain and controversial. We used degenerated oligonucleotide primer PCR (Dop-PCR), Dot-blot, and FISH to screen for a new marker of the St genome and to test the efficiency of this marker in the detection of the St chromosome at different ploidy levels. Signals produced by a new FISH marker (denoted St2-80) were present on the entire arm of chromosomes of the St genome, except in the centromeric region. On the contrary, St2-80 signals were present in the terminal region of chromosomes of the E, H, P, and Y genomes. No signal was detected in the A and B genomes, and only weak signals were detected in the terminal region of chromosomes of the D genome. St2-80 signals were obvious and stable in chromosomes of different genomes, whether diploid or polyploid. Therefore, St2-80 is a potential and useful FISH marker that can be used to distinguish the St genome from those of other genomes in Triticeae.

HSF1 transcription factor concentrates in nuclear foci during heat shock: relationship with transcription sites

1997 ◽  
Vol 110 (23) ◽  
pp. 2935-2941 ◽  
Author(s):  
C. Jolly ◽  
R. Morimoto ◽  
M. Robert-Nicoud ◽  
C. Vourc'h
Keyword(s):  
Transcription Factor ◽  
Transcription Factors ◽  
In Situ Hybridization ◽  
Heat Shock ◽  
Human Fibroblasts ◽  
Relative Distribution ◽  
Transcription Sites ◽  
Nuclear Foci ◽  
Hsp Genes

In this paper, we show that upon heat shock, HSF1 concentrates in the nucleus of diploid human fibroblasts in two large foci. The relative distribution of HSF1 nuclear foci and active heat shock protein (hsp) genes was investigated by combining fluorescence in situ hybridization (FISH) for the detection of hsp nuclear transcripts and immunofluorescence for the detection of HSF1. We show that the HSF1 foci are distinct from the sites of hsp70 and hsp90 genes transcription. This is the second report of ploidy-dependent foci of transcription factors that are independent of their specific transcription sites. However, the correlation between the number of HSF1 foci and the ploidy of the cells strongly supports the existence of a specific chromosomal target for HSF1 foci.

scholarly journals High-resolution FEISEM detection of DNA centromeric probes in HeLa metaphase chromosomes.

1995 ◽  
Vol 43 (4) ◽  
pp. 413-419 ◽  
Author(s):  
E Rizzi ◽  
M Falconi ◽  
R Rizzoli ◽  
B Baratta ◽  
L Manzoli ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
High Resolution ◽  
Centromeric Region ◽  
Secondary Electron Emission ◽  
Dna Probes ◽  
Dna Denaturation ◽  
Metaphase Chromosomes ◽  
Chromatin Arrangement ◽  
Fiber Organization

HeLa metaphase chromosome spreads were hybridized with centromeric biotinylated DNA probes and detected with gold-conjugated anti-biotin antibodies. Chromosomes were observed by an in-lens field emission scanning electron microscope (FEISEM), which permits detection of biological samples without any coating. DNA probes were well localized in the centromeric region of chromosomes and there was clear discrimination between 10 nm fibers that hybridized to DNA probes and those that did not hybridize. This approach shows that in situ hybridization can be directly visualized at the FEISEM level by evaluating only secondary electron emission, which allows physical localization of the hybridized probe with high resolution so that backscatter detection represents only a control. Because chromosomes maintain the 10-nm fiber organization after in situ hybridization procedures, our data suggest that this fiber represents the lowest order of chromatin arrangement that permits transitory DNA denaturation.

Labeling of the centromeric region on human chromosome 8 by in situ hybridization

1991 ◽  
Vol 87 (4) ◽  
Author(s):  
Heinz-UlrichG. Weier ◽  
JoeW. Gray ◽  
Hans-Dieter Kleine
Keyword(s):  
In Situ Hybridization ◽  
Human Chromosome ◽  
Centromeric Region ◽  
Chromosome 8
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