Blindness in uveitis results from degeneration of photoreceptor cells. This degeneration is attributed to macrophage infiltration and the subsequent generation of various inflammatory mediators, including the nitric oxide-derived highly reactive oxidant peroxynitrite, which is known to cause photoreceptor damage via nitration of tyrosine residues. Our preliminary data, however, shows the nitration commences in the early phase of EAU, prior to infiltration by macrophages. Moreover, we found that the nitration selectively involves the mitochondrial proteins. These novel findings implicate photoreceptor cell mitochondrial protein nitration as a central molecular event in the initiation of photoreceptor degeneration. This nitration is certainly not mediated by macrophages. We propose a hypothesis: """"""""Photoreceptor degeneration in autoimmune uveitis begins with selective mitochondrial protein nitration as a consequence of reactive nitric oxide species generation in the photoreceptor mitochondria."""""""" We will test this hypothesis with the following specific aims in animals with early EAU, prior to macrophage infiltration of the retina. 1. Evaluate nitric-oxide reactive species generation in the photoreceptor mitochondria (immunohistochemistry, immunoblotting, and mass spectrometry). 2. Evaluate mitochondrial protein nitration in the photoreceptors (immunohistochemical localization of nitrated proteins, immunoblotting of photoreceptor proteins, and capillary liquid chromatography-tandem mass spectrometry). 3. Evaluate functions of the photoreceptor mitochondria (oxygen consumption and activities of electron transport chain complexes, and mitochondrial transmembrane potential). To assure successful completion of the specific aims, we have assembled a multidisciplinary team of experts. Professor Terry Lee is an accomplished chemist with expertise in identifying nitrated proteins, and their site of nitration by mass spectrometry and other novel methods. Dr. Guey-Shuang Wu has expertise in PCR, Western blot, and separation of biological molecules. The principal investigator is experienced in the field of EAU, immunohistochemistry, free radical biology, and reactive nitric oxide species. Understanding the role of mitochondria in the pathogenesis of retinal degenerations could lead to the development of specific therapy to minimize mitochondrial generation of peroxynitrite.
| Kase, Satoru; Ishida, Susumu; Rao, Narsing A (2011) Increased expression of ?A-crystallin in human diabetic eye. Int J Mol Med 28:505-11 |
| Kase, S; Parikh, J G; Rao, N A (2009) Expression of heat shock protein 27 and alpha-crystallins in human retinoblastoma after chemoreduction. Br J Ophthalmol 93:541-4 |
| Rigas, Panayota K; Kase, Satoru; Rao, Narsing A (2009) Expression of alpha-crystallins in human sebaceous carcinoma of the eyelid. Eur J Ophthalmol 19:702-7 |
| Kase, Satoru; Parikh, Jignesh G; Rao, Narsing A (2009) Expression of alpha-crystallin in retinoblastoma. Arch Ophthalmol 127:187-92 |
| Khurana, Rahul N; Parikh, Jignesh G; Saraswathy, Sindhu et al. (2008) Mitochondrial oxidative DNA damage in experimental autoimmune uveitis. Invest Ophthalmol Vis Sci 49:3299-304 |
| Parikh, Jignesh G; Saraswathy, Sindhu; Rao, Narsing A (2008) Photoreceptor oxidative damage in sympathetic ophthalmia. Am J Ophthalmol 146:866-75.e2 |
| Saraswathy, Sindhu; Rao, Narsing A (2008) Photoreceptor mitochondrial oxidative stress in experimental autoimmune uveitis. Ophthalmic Res 40:160-4 |
| Rao, Narsing A; Saraswathy, Sindhu; Wu, Guey Shuang et al. (2008) Elevated retina-specific expression of the small heat shock protein, alphaA-crystallin, is associated with photoreceptor protection in experimental uveitis. Invest Ophthalmol Vis Sci 49:1161-71 |
| Rajendram, Ranjan; Saraswathy, Sindhu; Rao, Narsing A (2007) Photoreceptor mitochondrial oxidative stress in early experimental autoimmune uveoretinitis. Br J Ophthalmol 91:531-7 |
| Saraswathy, Sindhu; Wu, Gueyshuang; Rao, Narsing A (2006) Retinal microglial activation and chemotaxis by docosahexaenoic acid hydroperoxide. Invest Ophthalmol Vis Sci 47:3656-63 |
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