Analysis of genes and chromosomes by nonisotopic in situ hybridization

Lichter, Peter; Boyle, Ann; Cremer, Thomas; Ward, David C.

doi:10.1016/1050-3862(91)90005-c

Cited by 224 publications

(106 citation statements)

References 146 publications

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“…In contrast to the presumption that the decondensed genomic DNA contained in one chromosome is distributed throughout the nudeus (see, for example, Comings 1980), the DNA of individual chromosomes has been shown to occupy distinct territories (for review see Lichter et al 1991). This was concluded from experiments in which interphase cell nuclei were microirradiated to generate local chromatin damage.…”

Section: T Cremer Is At the Institut Fur Humangenetik Und Anthropolomentioning

confidence: 99%

“…Probe DNA was labeUed with biotin or digoxigenin by nick-translation as previously described (Lichter et al 1991). …”

Section: Probesmentioning

confidence: 99%

“…Chromosomal in situ suppression hybridization was performed as described (Lichter & Cremer 1992) using the pBS-libraries, derived from sorted human chromosomes, as probes and human Cot-1 DNA as competitor. Suspension cells were aUowed to adhere on poly L-lysine coated glass slides.…”

Section: Non-radioactive In Situ Hybridizationmentioning

confidence: 99%

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Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries

et al. 1993

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The nuclear topography of splicing snRNPs, mRNA transcripts and chromosome domains in various mammalian cell types are described. The visualization of splicing snRNPs, defined by the Sm antigen, and coiled bodies, revealed distinctly different distribution patterns in these cell types. Heat shock experiments confirmed that the distribution patterns also depend on physiological parameters. Using a combination of fluorescence in situ hybridization and immunodetection protocols, individual chromosome domains were visualized simultaneously with the Sm antigen or the transcript of an integrated human papilloma virus genome. Three-dimensional analysis of fluorescence-stained target regions was performed by confocallaser scanning microscopy. RNA transcripts and components of the splicing machinery were found to be generally excluded from the interior of the territories occupied by the individual chromosomes. Based on these findings we present a model for the functional compartmentalization of the cell nucleus. According to this model the space between chromosome domains, including the surface areas of these domains, defines a three-dimensional networklike compartment, termed the interchromosome domain (ICO) compartment, in wh ich transcription and splicing of mRNA occurs.

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Section: T Cremer Is At the Institut Fur Humangenetik Und Anthropolomentioning

confidence: 99%

“…Probe DNA was labeUed with biotin or digoxigenin by nick-translation as previously described (Lichter et al 1991). …”

Section: Probesmentioning

confidence: 99%

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Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries

et al. 1993

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“…Also, it was suggested that homologous chromosomes might be attached closer to each other than to nonhomologous chromosomes. The techniques to visualize and analyze (parts of) interphase chromosomes have evolved rapidly, revealing more accurate data on the position of chromosomes during interphase (for reviews see Manuelidis 1990;Jackson 1991;Lichter et al 1991).…”

Section: Introductionmentioning

confidence: 99%

The nuclear position of pericentromeric DNA of chromosome 11 appears to be random in GO and non-random in G1 human lymphocytes

et al. 1994

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The nuclear position of pericentromeric DNA of chromosome 11 appears to be random in G0 and non-random in G1 human lymphocytes Hulspas, R.; Houtsmuller, A.; Krijtenburg, P.-J.; Bauman, J.G.J.; Nanninga, N. Published in: Chromosoma DOI:10.1007/BF00352253 Link to publication Citation for published version (APA):Hulspas, R., Houtsmuller, A., Krijtenburg, P-J., Bauman, J. G. J., & Nanninga, N. (1994). The nuclear position of pericentromeric DNA of chromosome 11 appears to be random in G0 and non-random in G1 human lymphocytes. Chromosoma, 103, 286-292. DOI: 10.1007/BF00352253 General rightsIt is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. Download date: 12 May 2018Chromosoma ( Abstract. The nuclear topography of pericentromeric DNA of chromosome 11 was analyzed in G O (nonstimulated) and G 1 [phytohemagglutinin (PHA) stimulated] human lymphocytes by confocal microscopy. In addition to the nuclear center, the centrosome was used as a second point of reference in the three-dimensional (3D) analysis. Pericentromeric DNA of chromosome 11 and the centrosome were labeled using a combination of fluorescent in situ hybridization (FISH) and immunofluorescence. To preserve the 3D morphology of the cells, these techniques were performed on whole cells in suspension. Three-dimensional images of the cells were analyzed with a recently developed 3D software program (Interactive Measurement of Axes and Positioning in 3 Dimensions). The distribution of the chromosome 11 centromeres appeared to be random during the G O stage but clearly non-random during the G 1 stage, when the nuclear center was used as a reference point. Further statistical analysis of the G1 cells revealed that the centromeres were randomly distributed in a shell underlying the nuclear membrane. A topographical relationship between the centrosome and the centromeres appeared to be absent during the G O and G I stages of the cell cycle.

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“…Additionally, we analysed metaphase spreads from 7 melanoma cell lines with probes for c-myc, centromere of chromosome 8, and whole chromosome 8 (Pinkel et al, 1986;Lichter et al, 1991;Bar-Am et al, 1992).…”

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confidence: 99%

Extra c-myc oncogene copies in high risk cutaneous malignant melanoma and melanoma metastases

et al. 2001

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Summary Amplification and overexpression of the c-myc gene have been associated with neoplastic transformation in a plethora of malignant tumours. We applied interphase fluorescence in situ hybridization (FISH) with a locus-specific probe for the c-myc gene (8q24) in combination with a corresponding chromosome 8 α-satellite probe to evaluate genetic alterations in 8 primary melanomas and 33 advanced melanomas and compared it to 12 melanocytic nevi, 7 safety margins and 2 cases of normal skin. Additionally, in metaphase spreads of 7 melanoma cell lines a whole chromosome 8 paint probe was used. We investigated the functionality of the c-myc gene by detecting c-myc RNA expression with RT-PCR and c-myc protein by immunohistochemistry. 4/8 primary melanomas and 11/33 melanoma metastases showed additional c-myc signals relative to the centromere of chromosome 8 copy number. None of the nevi, safety margins or normal skin samples demonstrated this gain. In 2/7 melanoma cell lines (C32 and WM 266-4) isochromosome 8q formation with a relative gain of c-myc copies and a loss of 8p was observed. The highest c-myc gene expression compared to GAPDH was found in melanoma metastases (17.5%). Nevi (6.6%) and primary melanomas (5.0%) expressed the c-myc gene on a lower level. 72.7% of the patients with c-myc extra copies had visceral melanoma metastases (UICC IV), patients without c-myc gain in 35.0% only. The collective with additional c-myc copies also expressed the gene on a significantly higher level. These results indicate that a c-myc gain in relation to the centromere 8 copy number might be associated with advanced cutaneous melanoma.

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Analysis of genes and chromosomes by nonisotopic in situ hybridization

Cited by 224 publications

References 146 publications

Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries

Evidence for a nuclear compartment of transcription and splicing located at chromosome domain boundaries

The nuclear position of pericentromeric DNA of chromosome 11 appears to be random in GO and non-random in G1 human lymphocytes

Extra c-myc oncogene copies in high risk cutaneous malignant melanoma and melanoma metastases

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