A balanced chromosomal translocation affects the inner nucleus positioning of thederivative chromosomes or not.

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In the case of a patient with a disease associated balanced chromosomal rearrangements (DBCRs), one of the gene that disrupted by chromosomal rearrangements is possible to be confirmed as a responsible gene of the disease. If there were no known genes at the breakpoints, we have considered that the clinical features of the patient were not related to the chromosomal rearrangements. But, we speculated that it is possible that chromosomal rearrangements lead to change the inner nucleus spatial organization of the genome, and the breakpoint site affects the disease causing gene in such DBCRs patients. We analyzed a case of a balanced translocation: t(11; 22) (q23; q11) by FISH on three-dimensionally preserved cells (3D FISH) using several BAC clones, which were closely mapped the breakpoints. We devised a new design of the combination of the probes which can be recognized four different targeted chromosomes each other in a 3D preserved nucleus. We verified that newly-devised probe design is effective for this research. And, we established a protocol using direct-labeled FISH method instead of using previous time-consuming indirect-labeled method. We reviewed the appropriate images for this research, and we improved how to obtain the ideal images of the nucleus, how to operate 3D computer software, etc. Although, we could not finish 3D FISH analysis for many patients with DBCRs, and could not show by means of evidence of our hypothesis in two years, we established new strategy to detect four kinds of chromosomes involving translocation in a 3D preserved nucleus for our research purpose. These are the important products to propose the new mechanism of DBCRs and position effects.