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PubMed1976

The fluorescence localization of mouse satellite DNA in interphase nuclei.

Nuclei from various mouse tissues exhibit a pattern of fluorescence characteristic of the cell type when stained with the fluorescent compound Hoechst 33258. When such preparations are hybridized in situ with 3H-RNA complementary to the A-T rich satellite of mouse, it is clearly seen that only the fluorescent regions of the nuclei contain the satellite DNA. Thus Hoechst 33258 allows the precise localization of satellite DNA at all stages of the mouse cell cycle.

Animals↗
PubMed1995

Confirmation of the copy number of chromosome 1 in interphase nuclei from paraffin sections of breast tumours by fluorescence in situ hybridization.

Fluorescence in situ hybridization (FISH) was used to establish the copy number of chromosome 1 in a set of nine breast tumours in which the chromosome had previously been shown to have undergone a variety of rearrangements by loss of heterozygosity studies. In each case, FISH with satellite III DNA from chromosome 1q12 confirmed the results obtained by Southern hybridization. Importantly, in all five cases with rearrangements thought not to involve the centromeric region, FISH showed that the events had not disrupted the gross chromosome structure. This study highlights the potential of using the two techniques together to obtain a clearer picture of both large- and small-scale alterations to chromosomes in solid tumours.

Alleles↗
PubMed1996

Interphase Analysis of Chromosome 11 in Human Pituitary Somatotroph Adenomas by Direct Fluorescence In Situ Hybridization.

Chromosome 11 numerical abnormalities were detected by fluorescence in situ hybridization (FISH) technique in four surgically removed pituitary somatotroph adenomas from patients clinically associated with acromegaly. The tumors were diagnosed by histology. immunocytochemistry, and electron microscopy, and included two densely granulated somatotroph (DG-SM) and two sparsely granulated somatotroph (SG-SM) adenomas. For demonstration of chromosome 11, the direct FISH technique was applied on imprints from fresh adenoma tissue fixed in acetone using an a-satellite specific centromeric probe. The slides were studied with a fluorescence microscope and the percentages of positive nuclei with aberrant fluorescent signals were counted. All tumors exhibited numerical chromosomal abnormalities in 8-23% of their cell population and included nuclei containing 1-3 extra chromosome 11 copies. The SC-SM adenomas exhibited more prominent abnormalities compared with the DG-SM adenomas. We conclude that numerical chromosome 11 abnormalities represent a frequent event among somatotroph adenomas with a tendency for higher frequency in SC-SM adenomas.

Journal Article↗
PubMed1982

The lens epithelium and radiation cataract. IV. Ultrastructural studies of interphase death in the meridional rows.

The effect of radiation upon the differentiating meridional row cells of the rat lens epithelium was studied by electron microscopy. X-irradiation of rat lenses was shown to disrupt the nuclear and, in some instances, the cytoplasmic organization of these cells. Indications of damage, 4.5 h post-irradiation, are a disruption in the shape of the nucleus and its membranes. This is typified by a bulging or blebbing of the nucleus. At 12 h the affected nuclei deteriorate with obvious membrane breakdown. The nucleoplasm at this time may appear highly condensed and shrunken. Disruption of the plasma membrane is also evident. Based upon previous findings it is likely that the nuclear changes are independent of the processes involved in normal fiber denucleation.

Animals↗
PubMed2000

Interphase cytogenetics of bladder cancer progression: relationship between aneusomy, DNA ploidy pattern, histopathology, and clinical outcome.

In the present study, different stages of transitional cell carcinoma of the bladder were analyzed by fluorescent in situ hybridization, using probes specific for pericentromeric classical satellite. Seventy primary tumors were evaluated for chromosomes 1, 7, 9, 17, and ploidy by flow cytometry. The results were correlated, after a mean follow-up period, with ploidy, histopathological characteristics, recurrence, and progression. Firstly, our data demonstrated that the sensitivity of fluorescence in situ hybridization in detecting quantitative DNA aberrations exceeds that of flow cytometry. The frequency of chromosome 1 and 9 aberrations was not significantly different in diploid and aneuploid tumors of different stage and grade. In contrast, the chromosome 7 and 17 aneusomy showed greater differences between pT1 and pT2-3 tumors (P<0.032 and P<0.0006, respectively) than between stage pTa and pT1. An increasing number of aberrations was observed in all chromosomes examined from tumors of patients that afterwards underwent cystectomy and/or had recurrent tumors. This study indicates that fluorescence in situ hybridization could be used to detect genetic changes relevant to patient outcome. These genetic changes could identify patients at high risk of recurrence and possible progression.

Adult↗