应用DNA芯片技术作SNP单倍型连锁分析和关联分析、候选位置克隆策略寻找我国家族性开角型青光眼家系的相在致病基因；构建突变基因的原核、真核表达载体，转化大肠杆菌、转染小梁细胞，观察基因突变对蛋白功能特性、以及对体外培养小梁细胞功能特性的影响，以期阐明发性开角型青光眼的分子发病机理，为青光眼的基因治疗研究提供新的理论依据。

In this study, Using genome-wide scan and gene sequencing, sequence variation Pro370Leu in the TIGR/MYOC gene was identified. Using immunofluorescent microscopy, fluorescent of wild-type TIGR/MYOC gave rise to the formation of scattered punctuate in the cell; mutant TIGR/MYOC was accumulated as aggresome-like structure in the cell. In mutant TIGR/MYOC gene transfected cells, the F-actin was thicker and peripherally located. The vinculin was irregular and dispersed in the membrane with decreased number. By wound-healing method, the migration ability of HTM cells transfected with mutant TIGR/MYOC gene was decreased. Apoptosis cells analysis showed no significant difference between each group. In conclusion, Pro370Leu is the diease-causing mutation in GZ.1 pedigree. The possible pathogenesis of familial open-angle glaucoma was: the changes in primary structure of mutant TIGR/MYOC protein resulted in misfolded protein with incorrect conformation. The misfolded protein could not be secreted and accumulated in cytoplasma instead. The accumulation of the misfolded protein would led to the occurrence of POAG.

本研究应用全基因组扫描和基因测序法证实GZ.1开角型青光眼家系中TIGR/MYOC基因存在Pro370Leu突变；利用基因转染技术检测野生型以及突变型TIGR/MYOC蛋白的亚细胞器定位，细胞骨架蛋白的形态与分布，小梁细胞迁移能力，并检测细胞凋亡情况，经荧光免疫组化证实野生型TIGR/MYOC蛋白在小梁细胞内呈 点状、散在分布；而Pro370Leu突变型TIGR/MYOC蛋白是以积聚体形式存在于胞浆内；突变型TIGR/MYOC基因转染小梁细胞中微丝束粗大并向胞膜处边集，纽蛋白在核周的分布明显减少，而在胞膜处的分布则变得不连续、不规则；划痕实验显示突变TIGR/MYOC基因转染小梁细胞迁移率轻度下降。因此本研究证实GZ.1家系的致病基因为Pro370Leu突变型TIGR/MYOC基因，GZ.1家系可能的发病机理为：Pro370Leu突变型TIGR/MYOC蛋白一级结构的改变，导致蛋白质在内质网内错误折叠、构象发生严重错误，蛋白质无法正常分泌，在细胞内积聚，对小梁细胞造成毒性作用，最终导致了青光眼的发生。