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M. Akahori, M. Minami, M. Obazawa, S. Tomarev, N. Nakaya, Y. Miyake, T. Iwata; Characterization of Normal Tension Glaucoma Mouse Over Expressing Mutant OPTN (E50K) . Invest. Ophthalmol. Vis. Sci. 2006;47(13):206.
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© ARVO (1962-2015); The Authors (2016-present)
Recently optineurin (OPTN) was reported as responsible for primary open–angle glaucoma and normal tension glaucoma. This protein was previously characterized to interact with several proteins including small GTP–binding protein Rab8. We had previously reported that one of the mutations in OPTN (E50K) inhibited an interaction with Rab8 protein. To characterize the mutant OPTN (E50K) in relation with Rab8 interaction in vivo, transgenic mouse over expressing mutant OPTN (E50K) was developed.
Mouse OPTN was amplified by RT–PCR from C57BL/6 brain total RNA and E50K mutation and deletion of exon 5 were introduced by mutagenesis kit (Stratagene). All cDNA were inserted into transgenic construction vector (pBroad2). Transgenic mouse over expressing mutant OPTN (E50K) and mutant OPTN (deletion exon 5) were developed by YS Institute Inc. (Japan). All transgenic mice were examined by PCR method using genomic DNA extracted from mouse tail. Intraocular pressure was measured by Tonometer (Tonolab) and Optic fiber sensor (FISO technologies). Photograph of optic nerve head was observed by slit lamp with digital camera (Topcon). Histological characterization by HE staining and immunohistochemistry were performed. Apoptosis of retinal ganglion cells were detected by TUNEL kit (Roche).
Transgenic mouse over expressing OPTN (deletion exon 5) resulted as lethal expression. There was no difference in intraocular pressure between mutant OPTN (E50K) expressing mice and non–transgenic mice. Optic nerve in the surroundings of optic nerve head has decreased and retinal ganglion cells induced apoptosis were observed in mutant OPTN (E50K) expressing mouse.
Transgenic mouse over expressing wild type, E50K, and Exon 5 deleted OPTN was developed and characterized. Significant effect in retinal ganglion cells of E50K transgenic mouse was observed. This is the first POAG animal model developed by transgenic technology. Abnormality and OPTN–Rab8 interaction in vivo is under investigation.
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