Identification of Degeneration-Specific Genes in the Rcs Rat Retina By Subtractive Hybridization Techniques
Retinitis pigmentosa (RP) is one of the most frequent hereditary dystrophies. Multiple genetic defects have been found to be expressed in the photoreceptors and retinal pigment epithelium (RPE) cells of RP patients. The Royal College of Surgeons (RCS) rat is the animal model most commonly used to study RP. In the RCS rat, the photoreceptors begin to degenerate during the third postnatal week, and retinal degeneration continues toward the inner retina after their loss. We used a subtractive hybridization technique to identify differentially expressed genes that might be responsible for the induction of degenerative processes in the RCS retina. Until now, we have isolated about 30 clones with putative differentially expressed sequences. To date, two of these sequences were confirmed to be differentially expressed in the RCS rat but not in control rats. The method described here is therefore suitable for identifying differentially regulated genes, not only in RP retinas, but also in other retinal dystrophies and may help to further elucidate the pathophysiology of retinal diseases.
KeywordsRetinal Pigment Epithelium Cell Retinitis Pigmentosa Suppression Subtractive Hybridization Retinal Degeneration cDNA Array
Unable to display preview. Download preview PDF.
- 1.S. Merin, 1991, Inherited Eye Disese: Diagnosis and Clinical Management, Marcel Dekkcr, New York.Google Scholar
- 4.J.R. Heckenlively, 1988, Retinitis Pigmentosa, J. B. Lippincott, Philadelphia.Google Scholar
- 6.E. Zrenner, E. Apfelstedt-Sylla, K. Rüther, 1995, Clinical aspects: retinitis pigmentosa, in: Neurobiology and Clinical Aspects of the Outer Retina (M.B.A. Djamgoz, S.N. Archer, and S. Vallerga, eds.), pp. 447–460, Chapman & Hall, London.Google Scholar
- 7.E.L. Berson, 1994, Retinitis pigmentosa and allied diseases, in: Principles and Practice of Ophthalmology, 2 (D.M. Albert and F.A. Jakobiec, eds,), pp. 1214–1237, WB Saunders, Philadelphia.Google Scholar
- 8.M.A. Maw, B. Kennedy, A. Knight, R. Bridges, K.E. Roth, E.J. Mani, J.K. Mukkadan, D. Nancarrow, J.W. Crabb, M.J. Denton, 1997, Mutation of the gene encoding cellular retinaldehyde-binding protein in autosomal recessive retinitis pigmentosa, Nat. Genet. 17:198–200.Google Scholar
- 13.M.C. Bourne, D.A. Campell, K. Tansley, 1938, Hereditary degeneration of the rat retina, Br. J. Ophthalmol. 22:613–623.Google Scholar
- 19.K. Kohler, J. Hartmann, S. Fischer, E. Zrenner, 1997, Invest. Ophthalmol. 38:S38.Google Scholar
- 23.L. Diatchenko, Y.F. Lau, A.P. Campbell, A. Chenchik, F. Moqadam, B. Huang, S. Lukyanov, K. Lukyanov, N. Gurskaya, E.D. Sverdlov, P.D. Siebert, 1996, Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries, Proc. Natl. Acad. Sci. U.S.A. 93:6025–6030.PubMedCrossRefGoogle Scholar
- 24.J. Sambrook, E.F. Fritsch, T. Maniatis, 1989, Molecular Cloning*+Laboratory Manual, 2nd, Cold Spring Harbor Laboratory Press, New York.Google Scholar
- 29.P. Liang, L. Averboukh, A.B. Pardee, 1993, Distribution and cloning of eukaryotic mRNAs by means of differential display: refinements and optimization, Nucleic. Acids. Res. 21:3269–3275.Google Scholar
- 33.N.G. Gurskaya, L. Diatchenko, A. Chenchik, P.D. Siebert, G.L. Khaspekov, K.A. Lukyanov, L.L. Vagner, O.D. Ermolaeva, S.A. Lukyanov, E.D. Sverdlov, 1996, Equalizing cDNA subtraction based on selective suppression of polymerase chain reaction: cloning of Jurkat cell transcripts induced by phytohemaglutinin and phorbol 12-myristate 13-acetate, Anal. Biochem. 240:90–97.PubMedCrossRefGoogle Scholar