Three-dimensional chromatin distribution in neuroblastoma nuclei shown by confocal scanning laser microscopy

Brakenhoff, G. J.; Voort, H. T. M. van der; Spronsen, E. A. van; Linnemans, W. A. M.; Nanninga, N.

doi:10.1038/317748a0

Cited by 243 publications

(99 citation statements)

References 22 publications

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“…This was confirmed by using confocal microscopy. Confocal optics prevent most of the light that does not originate from the plane of focus from entering the detector (Minsky, 1961, Wilson and Sheppard, 1984, Brakenhoff et al, 1985. Therefore, confocal microscopes only examine a very thin optical section through the object and with the instrument employed in this study even brightly stained objects were no longer visible when the plane of focus was stepped by >0.8 pm.…”

Section: Internalisation Of Tenascinmentioning

confidence: 99%

Tenascin: a modulator of cell growth

End

Panayotou

Entwistle

et al. 1992

European Journal of Biochemistry

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The large, multidomain extracellular matrix protein tenascin displays a markedly restricted tissue distribution during embryogenesis and remains present only in a few adult tissues. The protein is re‐expressed, however, during wound healing and in the stroma of malignant tumours. While a variety of studies have dealt with the important role of tenascin in the development of neural and non‐neural tissues, there is growing evidence that tenascin expression may be associated with proliferation of cells lining these tissues. The presence of repeating domains in tenascin similar to those in epidermal growth factor prompted us to investigate the ability of tenascin to modulate the growth of different cell types. Tenascin was actually found to be mitogenic for several cell types. This mitogenic activity, however, appears to be associated with a region in the fibronectin type III domains. The mitogenic mechanism is clearly distinct from pathways used by peptide growth factors such as epidermal growth factor and platelet‐derived growth factor, which activate the intrinsic tyrosine kinase activity of their cell‐surface receptors. However, we show that this large extracellular matrix molecule is efficiently internalised and may be processed by responding cells.

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Section: Internalisation Of Tenascinmentioning

confidence: 99%

Tenascin: a modulator of cell growth

End

Panayotou

Entwistle

et al. 1992

European Journal of Biochemistry

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“…By repeating data collection in the x-y plane at successive depths in the specimen, 3-D information of high definition and resolution could be obtained. However, the first generation of such microscopes were not appropriate for biological experiments, with the breakthrough occurring in 1985 when Brakenhoff et al [35] …”

Section: Improving Imaging Resolutionmentioning

confidence: 99%

Seeing is believing: A focus on the contribution of microscopic imaging to our understanding of immune system function

Bajénoff

Germain

2007

Eur. J. Immunol.

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“…In this study we have attempted size and shape measurements of painted X-chromosome domains in cell nuclei of normal diploid, female, and male amniotic fluid cell cultures applying conventional fluorescence microscopy, confocal scanning laser fluorescence microscopy (CSLFM) (Brakenhoff et al, 1979(Brakenhoff et al, , 1985Cremer and Cremer, 1978;Stelzer et al, 1985Stelzer et al, , 1986; for review, see Shotton, 1989), and image analysis. Amniotic fluid cell nuclei resemble flat ellipsoids or flat cylinders with a n elliptic base.…”

Section: Introductionmentioning

confidence: 99%

Differences of size and shape of active and inactive X‐chromosome domains in human amniotic fluid cell nuclei

Bischoff¹,

Albers

Kharboush

et al. 1993

Microscopy Res & Technique

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It is a widely held belief that the inactive X-chromosome (Xi) in female cell nuclei is strongly condensed as compared to the largely decondensed active X-chromosome (Xa). We have reconsidered this problem and painted X-chromosome domains in nuclei of subconfluent, female and male human amniotic fluid cell cultures (46,XX and 46,XY) by chromosomal in situ suppression (CISS) hybridization with biotinylated human X-chromosome specific library DNA. FITCconjugated avidin was used for probe detection and nuclei were counterstained with propidium iodide (PI). The shape of these nuclei resembling flat ellipsoids or elliptical cylinders makes them suitable for both two-dimensional (2D) and three-dimensional (3D) analyses. 2D analyses of Xi-and Xa-domains were performed in 34 female cell nuclei by outlining of the painted domains using a camera lucida. Identification of the sex chromatin body in DAPI-stained nuclei prior to CISShybridization was confirmed by its colocalization with one of the two painted X-domains. In 31 of the 34 nuclei the area Axi for the inactive X-domain was smaller than the area A, , for the active domain (mean ratio Ax,/Ax, = 1.9 * 0.8 SD, range 1.0-4.3). The signed rank test showed a highly significant (P < .0001) difference both between A, , and Axi and between the ratios r(Xa) and r(Xi), calculated by dividing the maximum length L of each X-domain by its maximum width W. In most nuclei (26134) we found r(Xa)>r(Xi) demonstrating a generally more elongated structure of Xa. For 3D analysis a confocal scanning laser fluorescence microscope (CSLFM) was used. Ten to 20 light optical sections (PI-image, FITC-image) were registered with equal spacings (approx. 0.4 pm). A thresholding procedure was applied to determine the PI-labeled nuclear and FITC-labeled X-domain areas in each section. Estimated slice volumes were used to compute total nuclear and X-domain volumes. In a series of 35 female nuclei most domains extended from the top to the bottom nuclear sections. The larger of the two X-chromosome domains comprised (3.7 t 1.7 S.D.)% of the nuclear volume. A mean ratio of 1.2 k 0.2 SD (range 1.1-2.3) was found for the volumes of the larger and the smaller X-domains in these female nuclei. In a series of 27 male amniotic fluid cell nuclei the relative X-chromosome domain volume comprised (4.0 t 2.6 S.D.)%. These findings indicate that differences in the 3D expansion of active and inactive X-chromosome domains are less pronounced than previously thought. A current model suggests that chromosome domains consist of a compact core surrounded by loosely coiled outer chromatin fiber loops. The latter fraction may be considerably larger in Xa-as compared to Xi-domains. We suggest that the interactive outlining procedure used in the 2D analyses included the loosely structured domain periphery more accurately, while the threshold algorithm applied to light optical sections delineated the more compact core of the domains, leading to smaller and more similar volume estimates of Xa and Xi. Present limitat...

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Three-dimensional chromatin distribution in neuroblastoma nuclei shown by confocal scanning laser microscopy

Cited by 243 publications

References 22 publications

Tenascin: a modulator of cell growth

Tenascin: a modulator of cell growth

Seeing is believing: A focus on the contribution of microscopic imaging to our understanding of immune system function

Differences of size and shape of active and inactive X‐chromosome domains in human amniotic fluid cell nuclei

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