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

在患有与疾病相关的平衡染色体重排（DBCRs）的患者的情况下，被染色体重排破坏的基因之一可能被确认为该疾病的负责基因。如果在断点处没有已知的基因，我们认为患者的临床特征与染色体重排无关。但是，我们推测可能是染色体重排导致了基因组内部细胞核空间组织的改变，而断点位点影响了这类dbcr患者的致病基因。我们分析了一个FISH在三维保存细胞（3D FISH）上的平衡易位：t(11; 22) （q23; q11）的案例，使用了几个BAC克隆，这些克隆与断点密切相关。我们设计了一种新的探针组合，可以在一个三维保存的细胞核中识别四个不同的目标染色体。我们验证了新设计的探针是有效的。并且，我们用直接标记的FISH方法代替了之前耗时的间接标记方法，建立了一个协议。我们回顾了适合本研究的图像，改进了如何获得理想的细胞核图像，如何操作三维计算机软件等。虽然我们无法完成许多dbcr患者的3D FISH分析，并且在两年内无法通过证据证明我们的假设，但我们建立了新的策略，在3D保存的细胞核中检测四种涉及易位的染色体，以达到我们的研究目的。这些都是提出dbcr新机制和位置效应的重要产物。

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.