Investigative ophthalmology & visual science · 1999
Beta-arrestin-related proteins in ocular tissues.
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Nicolas-Léveque C, Ghedira I, Faure J P, Mirshahi M
Аннотация
PURPOSE: Proteins of the arrestin family contribute to the regulation of G-protein-mediated transduction. In this study, the presence of beta-arrestins in ocular tissues was investigated.
METHODS: Mouse monoclonal and rabbit polyclonal antibodies were raised against the peptide Val-Asp-Thr-Asn-Ile-Leu-Glu-Leu-Asp-Thr-Asn-Asp-Asp-Asp-Ile, a sequence present in beta-arrestins 1 and 2 but absent from visual arrestin. These antibodies were used for the immunohistologic detection of beta-arrestins in parafin sections of rodent eyes fixed in Bouin's solution. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of RNA from bovine retina, retinal pigmented epithelial (RPE) cells, lens epithelial cells, and human corneal fibroblasts was performed using beta-1 arrestin primers.
RESULTS: In the eye, bet-arrestin staining predominated in RPE, inner segments of photoreceptors, synaptic spherules of rods, inner plexiform layer and ganglion cell fibers, epithelial cells from ciliary body, and vessels. RT-PCR amplified a 480 bp product, corresponding to the predicted length. The sequence of PCR products from bovine retina and RPE cells was identical with the bovine beta-arrestin mRNA.
CONCLUSIONS: beta-arrestins were detected in several ocular tissues. In photoreceptor cells, their specific localization in the synaptic terminals and plexiform layer suggests a role of beta-arrestin in synaptic transmission. In other ocular tissues, the presence of beta-arrestin may be related either to adrenergic signal transduction or to signal transduction mediated by other G-protein-coupled receptors.
METHODS: Mouse monoclonal and rabbit polyclonal antibodies were raised against the peptide Val-Asp-Thr-Asn-Ile-Leu-Glu-Leu-Asp-Thr-Asn-Asp-Asp-Asp-Ile, a sequence present in beta-arrestins 1 and 2 but absent from visual arrestin. These antibodies were used for the immunohistologic detection of beta-arrestins in parafin sections of rodent eyes fixed in Bouin's solution. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of RNA from bovine retina, retinal pigmented epithelial (RPE) cells, lens epithelial cells, and human corneal fibroblasts was performed using beta-1 arrestin primers.
RESULTS: In the eye, bet-arrestin staining predominated in RPE, inner segments of photoreceptors, synaptic spherules of rods, inner plexiform layer and ganglion cell fibers, epithelial cells from ciliary body, and vessels. RT-PCR amplified a 480 bp product, corresponding to the predicted length. The sequence of PCR products from bovine retina and RPE cells was identical with the bovine beta-arrestin mRNA.
CONCLUSIONS: beta-arrestins were detected in several ocular tissues. In photoreceptor cells, their specific localization in the synaptic terminals and plexiform layer suggests a role of beta-arrestin in synaptic transmission. In other ocular tissues, the presence of beta-arrestin may be related either to adrenergic signal transduction or to signal transduction mediated by other G-protein-coupled receptors.
Предметные рубрики MeSH
Amino Acid SequenceAnimalsAntibodies, MonoclonalArrestinsBlotting, WesternCattleCells, CulturedCiliary BodyDNA PrimersElectrophoresis, Agar GelFluorescent Antibody Technique, IndirectHumansMiceMolecular Sequence DataPeptide FragmentsPigment Epithelium of EyeRabbitsRetinaReverse Transcriptase Polymerase Chain Reactionbeta-Arrestins