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Akiko Maeda, Hiroshi Ohguro, Tadao Maeda, Takashi Nakagawa, Yoshio Kuroki; Low Expression of αA-Crystallins and Rhodopsin Kinase of Photoreceptors in Retinal Dystrophy Rat. Invest. Ophthalmol. Vis. Sci. 1999;40(12):2788-2794.
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© ARVO (1962-2015); The Authors (2016-present)
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 αA-crystallin. Reverse
transcription–polymerase chain reaction confirmed selective low levels
of mRNA expressions of αA-crystallins and RK in RCS rats.
conclusions. This study demonstrates that decreased expression of αA-crystallins
and RK in RCS rats, may have significant roles in the development of
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