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I. Kimura, H. Okamoto, Z.-L. Chi, M. Akahori, M. Minami, M. T. Suzuki, T. Iwata; Analysis of Colocalization of Rab8 and ERM Family in the Ocular Ciliary Body. Invest. Ophthalmol. Vis. Sci. 2009;50(13):897.
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© ARVO (1962-2015); The Authors (2016-present)
To perform proteome analysis of ocular ciliary body and investigate localization of several proteins supposed to relate to the pathological mechanism of glaucoma identified from the proteomics.
(1)Ciliary bodies from healthy cynomolgus monkey (Macaca fascicularis) eyes were obtained and processed within 4 hours after death. In order to investigate proteins localized in nonpigment and pigment epithelium more in detail, these tissues were isolated from cynomolgus monkey eye with stromal tissue left behind. Protein lysates were prepared, and tryptic peptides were extracted and analyzed by liquid chromatography coupled to tandem electro spray mass spectrometry(LCQ Deca XP plus, Thermo Fisher Scientific Inc.). Database searches were performed with the asistance of protein search program(Bio Works 3.3.1;Thermo Fisher Scientific Inc.). (2)Rab8, a member of Ras-related protein, which has interaction with Optineurin, and ERM family(Ezrin, Radixin, Moesin), which is microvillar core protein and linker protein of actin filament and plasma membrane, were identified from the study. Localization of these proteins in the ciliary body of cynomolgus monkey and mouse was investigated by immunostaining.
(1)374 proteins were identified from the ciliary epithelium, and 318 proteins from stromal tissue. Total number of proteins identified was 550. (2)Rab8, Ezrin, Radixin, and Moesin were proved to localize in the ciliary body of cynomolgus monkey and mouse by immunostaining. Colocalization of Rab8 and Moesin, Rab8 and Radixin was also proved by double immunostaining.
These results provide a starting point for building a database of the ocular ciliary body proteome, and for the solution of pathological mechanism of glaucoma in the interaction of Rab 8 and ERM family.
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