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R. Iwakiri, K. Kobayashi, H. Kobayashi, S. Okinami; Distribution of a Retinal Pigment Epithelium-specific Gene Product in Chick Embryonic Eyes and Mouse Eyes . Invest. Ophthalmol. Vis. Sci. 2003;44(13):397.
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Purpose: pP344 gene, which was isolated from chick embryonic retinal pigment epithelium (RPE), is specifically expressed in RPE in chick eyes. The expression is closely associated with in vivo differentiation and in vitro transdifferentiation to lens cells. The pP344 has several domains analogous to serine protease inhibitor. We raised an antibody for the chick pP344 gene product (PP344 protein) to examine the localization of the protein in the eye. Methods:From the deduced amino acid sequence of pP344, we synthesized a 15-amino-acid peptide, and immunized rabbits with the peptide. ELISA was used to assess the titer of the antiserum against the PP344 protein. Immunohistochemical study was employed to investigate the distribution of the PP344 protein in 9-day-old chick embryonic eyes and 8-week-old mouse eyes. We carried out Western blotting using cell extract of neural retina and RPE cells of chick embryos. Results: Immunohistochemical study showed that the significant staining of the antibody to the PP344 protein distributed in RPE and the entire neural retina in chick embryonic eyes. In 8-week-old mouse eyes, the immunostaining was localized to RPE and photoreceptor cell layer to ganglion cell layer in mouse eye. Western blotting analysis detected the band of 65kDa in cell extract of RPE cells and neural retina. Conclusions: This study showed that the PP344 protein was localized in RPE and neural retina, indicating that PP344 protein may be secreted to the neural retina side. The presence of the PP344 protein in neural retina suggests that this protein may play an important role in functions of neural retina.
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