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

在与疾病相关的平衡染色体重排(DBCRs)患者中，被染色体重排破坏的基因之一可能被确认为该疾病的责任基因。如果断裂点没有已知的基因，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，染色体重排可能导致基因组内核空间组织的改变，而断裂点位置影响这类DBCRs患者的致病基因。我们分析了一个平衡易位的案例：T(11；22)(q23；q11)通过FISH在三维保存的细胞(3D FISH)上使用了几个BAC克隆，这些克隆紧密地映射了断裂点。我们设计了一种新的探针组合设计，可以在3D保存的细胞核中相互识别四个不同的靶染色体。验证了新设计的探头设计对本研究的有效性。并且，我们建立了一种直接标记FISH方法来代替以前耗时的间接标记方法的协议。我们回顾了适合本研究的图像，并改进了如何获得理想的细胞核图像，如何操作3D计算机软件等。尽管我们无法完成许多DBCRs患者的3D FISH分析，也无法在两年内通过我们的假设证据显示，但为了我们的研究目的，我们建立了新的策略来检测3D保存的细胞核中涉及易位的四种染色体。这些都是提出DBCRs和位置效应新机制的重要成果。

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.