Interphase FISH

RAMIREZ, M.J., SURRALLES, J., GALOFRE, P, CREUS, A., MARCOS, R., Radioactive iodine induces clastogenic and age-dependent aneugenic effects in lymphocytes of thyroid cancer patients as revealed by interphase FISH, Mutagenesis 12 (1997) 449-455. 

INTERPHASE FISH Several recurrent genetic abnormalities identified in small cell lymphocytic lymphoma/chronic lymphocytic leukemia have had a direct relationship to disease progression. These abnormalities are ... 

DeLeeuw B, Suijkerbuijk RF, Olde-W huis D, et al. Distinct Xpll.2 breakpoint regions in synovial sarcoma revealed by metaphase and interphase FISH Relationship to histologic sub-types. Cancer Genet Cytogenet. 1994 73 89-94. 

Overrepresentation of the chromosome 12p region is a consistently present structural chromosomal aberration in GCT. Isochromosome 12p is present in up to 80% of GCTs including extratesticular tumors. Therefore the identification of isochromosome 12p by interphase FISH-based cytogenetics is occasionally resorted to in cases where the previously mentioned IHC approach fails to resolve the differential diagnosis of GCT versus somatic carcinoma. 

Oliva E, de Leval L, Soslow RA, et al. High frequency of JAZFl-JJAZl gene fusion in endometrial stromal tumors with smooth muscle differentiation by interphase FISH detection. Am J Surg Pathol. 2007 31 1277-1284. 

The applications of FISH for chromosome-based studies involve the use of chromosome spreads, chromosomes of tissue sections, and interphase nuclei. Other applications include the study of adventitious organisms (2) and messenger RNA synthesis (3). The applications, especially to chromosome spreads, are sometimes enhanced by exploiting the opportunity presented by FISH for double or triple labeling. 

Table 1 lists representative examples of the application of in situ hybridization, including FISH, to studies that use chromosome preparations and interphase nuclei (4,5). FISH is superior to in situ hybridization with an enzymatic probe because it provides finer resolution and higher signal intensity. This is especially important when the localization of a gene within a specific chromosome band is to be established. 

American College of Medical Genetics, Prenatal interphase fluorescence in situ hybridization (FISH) policy statement. Am. J. Hum. Genet. 53(2), 526-527 (1993). 

Gene amplification of the BCR-ABL 1 kinase may be associated with development of IM resistance. The presence of multiple copies of the BCR-ABL 1 gene in interphase nuclei can be demonstrated by fluorescence in situ hybridization (FISH) (40,42). In one series, more than half of the patients with IM resistance had evidence of clonal evolution with the development of additional chromosome abnormalities. Paired cytogenetic analyses performed at the begiiming of IM therapy and at the time of resistance demonstrated this evolution. 

Walter J, Joffe B, Bolzer A, Albiez H, Benedetti PA, Muller S, Speicher MR, CremerT, Cremer M, Solovei I (2006) Towards many colors in FISH on 3D-preserved interphase nuclei. Cytogenet Genome Res 114 367-378. 

For numerical chromosomal alterations, it is possible to analyze interphase cells using FISH with whole chromosome paints, or chromosomal specific probes. Thus, it is feasible to assess nondisjunctional events in any cell type, including target cells for tumorigenesis. Other fluorescence-based methods described in the section on in vitro methods (Section 25.3.4b) can be used in vivo, but they are of rather limited value for basic mutagenicity assays. 

PI clones Genomic sequences 90 kb Genes Mapping Interphase cytogenetics Usually make good FISH probes High... 

It will continue to be of considerable importance to establish a specific role for chromosome loss in tumor development. The analysis of aneuploidy in interphase cells of solid tumors using FISH will be greatly advantageous in this respect. For cancer risk assessment purposes, results from aneuploidy assays can be considered particularly useful when the mode of action of a chemical is known to result in chromosome loss or nondisjunction. 

Fluorescence in situ hybridization (FISH) is a common technique used to detect gene rearrangements, regions of chromosome deletion and amplification, and numerical chromosomal abnormalities. It utilizes fluorescently labeled DNA probes that bind to homologous chromosomal regions and can assay interphase nuclei. FISH can be performed on a variety of tissue specimens including frozen tissue or FFPE tissue sections, touch preparations from fresh or frozen tissue samples, cultured cells, and cytologic smears. 

Fluorescence in situ hybridization (FISH) has been usually performed on mitotic chromosomes or in interphase nuclei. The reagents for FISH are routinely used with mitotic chromosomes. The method reported here is based on protocols used in David C. Ward s lab (personal communication) at Yale University (Department of Genetics, School of Medicine, New Haven, CN). General methods for FISH in meiotic cells were reviewed by Spyropoulos and Moens (1994). 

---