Molecular genetics and cell biology of a glaucoma gene and the gene product

青光眼基因及其基因产物的分子遗传学和细胞生物学

• 批准号: 11671725
• 负责人: SUZUKI Yasuyuki
• 金额: $ 2.24万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671725/
• 关键词: glaucoma; gene mutations; myocilin; phenotype; bovine trabecular meshwork culture cells; in situ hybridization; cDNA cloning; steroid

Mutations in myocilin gene were screened in more than 150 primary open-angle glaucoma patients with single strand conformation polymorphism and direct sequencing. There was no patient who had family history of glaucoma and a mutation in myocilin gene but one pedigree in addition to the ones we previously reported had a mutation in myocilin gene at the 1109th nucleotide pair. The mutation was C1109T which resulted in Pro370Leu amino acid mutation. The phenotype of the patients was almost the same as the one we previously reported : the patients developed very high intraocular pressure earlier than 20 years old and had a juvenile onset type open-angle glaucoma. For the purpose of investigating the function of myocilin protein, we cloned bovine myocilin cDNA using the cultured bovine trabecular meshwork cells. According to the determined cDNA sequence, we made nucleotide probes and investigated distribution of myocilin in bovine eye by in situ hybridization. Using semi-quantitive PCR and realtime quantitive PCR, we investigated the change of expression of myocilin mRNA by steroid stimulation. The primer pairs and fluorescein probes were constructed based on the determined bovine myocilin cDNA sequence. As a result, dexamethason treatment induced myocilin mRNA expression dose-dependently.

在150多个单链构象多态性和直接测序的150多名原发性开式青光眼患者中筛选肌动蛋白基因的突变。没有患者具有青光眼家族史和肌动蛋白基因的突变，但是除了我们先前报道的那些血统，在第1109核苷酸对的肌动蛋白基因中有突变。该突变为C1109T，导致Pro370Leu氨基酸突变。患者的表型几乎与我们先前报道的患者相同：患者早于20岁以前的眼内压很高，并且有少年发作类型的开态角化青光眼。为了研究肌动蛋白蛋白的功能，我们使用培养的牛小梁网络将牛肌动蛋白cDNA克隆。根据确定的cDNA序列，我们通过原位杂交进行了核苷酸探针，并研究了牛眼中肌动蛋白的分布。使用半质量PCR和实时数量PCR，我们通过类固醇刺激研究了肌动蛋白mRNA的表达变化。基于确定的牛肌动蛋白cDNA序列构建引物对和荧光素探针。结果，右美森治疗诱导肌动蛋白mRNA表达剂量依赖性。

项目成果

Taniguchi F, Suzuki Y, Kurihara H, Kurihara Y, Kasai H, Shirato S, Araie M: "Molecular cloning of the bovine MYOC and induction of its expression in trabecular meshwork cells."Invest Ophthalmol Vis Sci. 41. 2070-2075 (2000)

Taniguchi F、Suzuki Y、Kurihara H、Kurihara Y、Kasai H、Shirato S、Araie M：“牛 MYOC 的分子克隆及其在小梁网细胞中的表达诱导。”Invest Ophasemol Vis Sci。

Fumiko TANIGUCHI et. al.: "Molecular cloning of the hovine MYOC and induction of its expression in trahecnlar meshwork cells."Inrest Ophthalmol Vis Sci. 41. 2070-2075 (2000)

谷口富美子 等

Fumiko TANIGUCHI et al.: "Molecular cloning of the borine MYOC and induction of its expression in trabecular moshword cells"Invest Ophthalmol Vis Sci. 41. 2070-2075 (2000)

Fumiko TANIGUCHI 等人：“硼素 MYOC 的分子克隆及其在小梁 moshword 细胞中表达的诱导”Invest Ophasemol Vis Sci。

Taniguchi F, Suzuki Y, Shirato S, Araie M: "The Gly367Arg mutation in the myocilin gene causes adult-onset primary open-angle glaucoma."Jpn J Ophthalmol. 44. 445-448 (2000)

Taniguchi F、Suzuki Y、Shirato S、Araie M：“myocilin 基因中的 Gly367Arg 突变导致成人发病的原发性开角型青光眼。”Jpn J Ophthalmol。

Fumiko TANIGUCHI et al.: "The Gly367Arg mntation in the myocilin gene causes adult-onset primary open-angle glancoma"Jpn J Ophthalmol. 44. 445-448 (2000)

Fumiko TANIGUCHI 等人：“myocilin 基因中的 Gly367Arg 表达导致成人发病的原发性开角型青光眼”Jpn J Ophamol。