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

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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.