Correlation of cytogenetic and fluorescence in situ hybridization (FISH) studies in normal and gliotic brain.

Abstract

In vitro tissue culturing, for karyotype analysis, may introduce artifacts confounding the cytogenetic evaluation of tissues with low baseline proliferative activity. Utilizing a panel of fluorescence in situ hybridization (FISH) probes for chromosomes 7, 8, 9, 10, 12, 17, 18, X, and Y, we compared the results of FISH analysis of non-tumorous normal (11 patients) and gliotic (10 patients) brain tissue touch preparations with those of cytogenetic evaluation performed on short-term primary cultures of the same material. We found a significant rate of apparent monosomy of chromosomes 8 and 17 by FISH analysis, with no corresponding clonal chromosomal loss detected by karyotype evaluation. These monosomy rates were significantly lower in gliotic than in normal brain tissue, and image analysis suggested that this apparent monosomy was due to interphase pairing of homologous centromere signals. Two distinct Y-chromosome signals were seen in 9.4% of nuclei by FISH, with 3 of 15 males displaying disomy Y rates over 15%. Disomy Y rates correlated approximately with age and clonal disomy Y was seen in the karyotype of one of these specimens. Karyotype analysis demonstrated loss of a sex chromosome in 6 specimens, while no sex chromosome nullisomy was detected by FISH. FISH is a valuable adjunct to the cytogenetic evaluation of tissues with low baseline proliferative activity. The differences in relative monosomy rates between normal and gliotic brain suggest that alterations in nuclear architecture and/or DNA sequence accompany the transition from normal to reactive glia.