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(Investigative Ophthalmology and Visual Science. 1999;40:2788-2794.)
© 1999 by The Association for Research in Vision and Ophthalmology, Inc.

Low Expression of {alpha}A-Crystallins and Rhodopsin Kinase of Photoreceptors in Retinal Dystrophy Rat

Akiko Maeda1, Hiroshi Ohguro1, Tadao Maeda1, Takashi Nakagawa1 and Yoshio Kuroki2

From the Departments of 1 Ophthalmology and 2 Biochemistry, Sapporo Medical University School of Medicine, Japan.

PURPOSE. The Royal College of Surgeons (RCS) rat has been extensively characterized as a model for inherited retinal dystrophy such as retinitis pigmentosa. In the present study, compositions of retinal proteins were compared between RCS (rdy-/-) and control (rdy+/+ ) rats during progression of the disease to understand the molecular pathologic course of the retinal degeneration.

METHODS. Protein mapping was performed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) or two-dimensional (2D)-PAGE using whole retinas or rod outer segments (ROS) obtained by a sucrose-density gradient centrifugation method from RCS or control rats at the age of 3 to 8 weeks.

RESULTS. 2D-PAGE showed that retinal proteins of RCS rats were generally less abundant than those of the control animals and that the difference became more evident with aging. However, no significant difference was observed in the protein-mapping patterns in 2D-PAGE between RCS and control rats in any ages tested. Analysis by SDS-PAGE of ROS proteins and by western blot using antibodies against opsin, rhodopsin kinase (RK), recoverin, or arrestin demonstrated that a 20-kDa protein and RK were selectively less abundant in RCS than in control rats. Edman sequence analysis of the proteolytic peptides obtained by in-gel digestion of the corresponding protein band using endoproteinase Lys C identified the 20-kDa protein as {alpha}A-crystallin. Reverse transcription–polymerase chain reaction confirmed selective low levels of mRNA expressions of {alpha}A-crystallins and RK in RCS rats.

CONCLUSIONS. This study demonstrates that decreased expression of {alpha}A-crystallins and RK in RCS rats, may have significant roles in the development of retinal dystrophy.




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H. Yamazaki, H. Ohguro, T. Maeda, I. Maruyama, Y. Takano, T. Metoki, M. Nakazawa, H. Sawada, and M. Dezawa
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Copyright © 1999 by the Association for Research in Vision and Ophthalmology