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      Expression of TGF-β superfamily receptors in the retinal pigmented epithelium Translated title: Expressão de receptores da superfamília de TGF-β no epitélio pigmentário da retina

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          Abstract

          SUMMARY Background: Retinal pigment epithelial (RPE) cells play an important role in the inflammatory response of the eye. Trans-forming growth factor-beta (TGF-β) and other members of the TGF-β superfamily are described to regulate some RPE cells functions. In this study the expression of TGF-β superfamily receptors in RPE cells at mRNA level was investigated Methods: RT-PCR technique was performed using mRNAs from D407 RPE cells (human RPE cell line) and HaCatTcells (human keratinocyte cell line used as positive control). Results: Expression of 6 type 1 receptors (TGF-β type I receptor, ALK-1, Activin type I receptor, activin type IB receptor, BMP type SA receptor, BMP type IB receptor), and 4 type II receptors (TGF-β type lI receptor, activin type II receptor, activin type IIB receptor, BMP type II receptor) were studied. The results demonstrated that TGF-β, activins and BMPs express their own specific receptors at mRNA level. Conclusions: The present study suggests that TGF-β superfamily members can exert effects on D407 RPE cells through their specific receptors.

          Translated abstract

          RESUMO Objetivo: O epitélio pigmentário da retina (EPR) desempenha um importante papel na resposta inflamatória ocular. "Transforming growth factor-beta" (TGF-β) e outros membros de sua superfamília têm sido descritos como reguladores de certas funções do EPR. Neste estudo, os autores investigaram a expressão de receptores da superfamília de TGF-β superfamily nas células do EPR a nível de RNA mensageiro. Métodos: Técnica de RT-PCR foi usada com RNA mensageiro de D407 (linhagem de células do EPR humano) e HaCatT (linhagem de queratócitos humanos usados como controle positivo). Resultados: A expressão de 6 receptores tipo I (TGF-β receptor tipo I, ALK-I, activina receptor tipo I, activina receptor tipo IB, BMP receptor tipo IA, BMP receptor tipo IB), e 4 receptores tipo II (TGF-β receptor tipo II, activina receptor tipo II, activina receptor tipo IIB, BMP receptor tipo II) foi investigada. Os resultados demonstraram que TGF-β, activinas e BMPs expressam receptores específicos a nível de RNA mensageiro em células do EPR. Conclusões: O presente estudo sugere que membros da superfamília de TGF-β podem exercer efeitos nas células de EPR D407 através de seus receptores específicos.

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          Most cited references 21

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          Pigment epithelium proliferation in retinal detachment (massive periretinal proliferation).

           R Machemer,  H Laqua (1975)
          Cell proliferation and membrane formation found in the posterior parts of the eye after production of retinal detachment in owl monkey eyes had either the characteristics of macrophages or of membrane-forming epithelial cells. The proliferating cells originated from pigment epithelium. These membranes caused folding of the retina and strand formation in the vitreous cavity. Since cell proliferation was present on all available surfaces, such as the upper and under surface of the retina, and along vitreous surfaces, we propose to call this entity "massive periretinal proliferation" (MPP).
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            A human retinal pigment epithelial cell line that retains epithelial characteristics after prolonged culture.

            A spontaneously arising, apparently transformed, cell line has been cloned from a primary culture of human retinal pigment epithelial (RPE) cells and has been subcultured more than 200 times. The similarities of these cells to human RPE cells in vivo have been determined. The structure of the transformed cells has been determined by light and electron microscopy and by immunocytochemistry using antibodies that detect cytoskeletal and other proteins. The ability of the cell line to bind and phagocytose photoreceptor material has also been assessed by fluorescence and electron microscopy. The metabolism of all-trans-retinol has been investigated by incubation of the cells with 3H-all-trans-retinol and analysis of the metabolic products by high-performance liquid chromatography. The transformed cells possess an epithelial cobblestone morphology with intercellular junctional complexes containing N-cadherin. The cytoskeleton of these cells comprises cytokeratins that are characteristic of epithelial cells, together with actin, spectrin, and vimentin. The keratins expressed are those typical of RPE cells. The cells also express cellular retinaldehyde binding protein and retinol dehydrogenase activity but do not express retinoid isomerase or lecithin retinol acyl transferase activities. These cells also exhibit phagocytic activity. This cell line retains many of the metabolic and morphologic characteristics of RPE cells in vivo although there are some differences, particularly the loss of some enzymatic activities and cytoskeletal polarization. These cells should be useful in further studies of RPE cell metabolism and other functions.
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              Transforming growth factor beta 2 is the predominant isoform in the neural retina, retinal pigment epithelium-choroid and vitreous of the monkey eye.

              Several techniques were utilized to assess the levels, disposition and cellular sources of isoforms 1 and 2 of transforming growth factor beta (TGF-beta) in the posterior pole of the monkey eye. Freshly dissected tissues, as well as the saline vehicles in which dissections were performed, were analysed by sandwich enzyme-linked immunosorbent assay. In all tissues TGF-beta 2 was the predominant isoform, with beta 2:beta 1 ratios of 6:1 for neural retina (as ng g-1) and 425:1 for vitreous (as pmol l-1). Retinal pigment epithelium (RPE)-Bruch's membrane-choroid complex contained approximately 10 times the amount of both TGF-beta isoforms as neural retina. For first passage cultures of monkey RPE, TGF-beta 2, but not TGF-beta 1, accumulated over time in conditioned media samples. Immunoreactivity for TGF-beta 2 was detected both in tissue sections of posterior pole, specifically in rod outer segments and RPE, and also in the first passage cultures of RPE. Antibodies to specific peptide sequences of both isoforms localized TGF-beta to the outer segments of rod photoreceptors. The apparent sequestration of TGF-beta 2 in photoreceptor outer segments, as well as the in vitro evidence for possible synthesis and release by RPE, suggest that TGF-beta 2 is an important modulator of visual function acting at the retina-RPE interface.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                abo
                Arquivos Brasileiros de Oftalmologia
                Arq. Bras. Oftalmol.
                Conselho Brasileiro de Oftalmologia (São Paulo, SP, Brazil )
                0004-2749
                1678-2925
                April 1998
                : 61
                : 2
                : 148-151
                Affiliations
                Tokyo orgnameJapanese Foundation for Cancer Research orgdiv1Cancer Institute orgdiv2Department of Biochemistry Japan
                Tokyo Kanto orgnameUniversity of Tokyo orgdiv1Faculty of Medicine orgdiv2Department of Ophthalmology Japan
                São Paulo orgnameFederal University of São Paulo orgdiv1Paulista School of Medicine Brazil
                Article
                S0004-27491998000200148
                10.5935/0004-2749.19980071

                This work is licensed under a Creative Commons Attribution 4.0 International License.

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                Figures: 0, Tables: 0, Equations: 0, References: 22, Pages: 4
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