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Regulation of RPE intercellular junction integrity and function by hepatocyte growth factor.
Invest Ophthalmol Vis Sci. 2002 Aug; 43(8):2782-90.IO

Abstract

PURPOSE

To evaluate the effect of hepatocyte growth factor (HGF) on the integrity and function of tight junctions and adherens junctions in the retinal pigment epithelial (RPE) monolayer.

METHODS

Fresh bovine eyes were dissected to obtain 2- to 3-mm(2) explants of intact RPE with underlying choroid and sclera. Explants were cultured with or without HGF (20 ng/mL) for various periods (20 minutes to 72 hours). Junction integrity was assessed by transmission and scanning electron microscopy; localization, expression, and phosphorylation of junction proteins; and measurement of transepithelial resistance (TER), diffusion of fluorescent labeling in the plasma membrane, and the migration of RPE cells from the monolayer.

RESULTS

Untreated explants consisted of polarized cells with apical microvilli and well-developed tight and adherens junctions. After HGF treatment, the explants showed loss of tight and adherens junctions ultrastructurally, diffusion of fluorescent label from apical to lateral membrane domains, and increased chemotactic migration of RPE cells from the monolayer. Primary cultures of confluent RPE cells showed a progressive decrease in TER. Western blot analysis showed rapid tyrosine phosphorylation of ZO-1, occludin, and beta-catenin within 20 minutes of stimulation. There was a marked loss of ZO-1 protein within 1 hour of HGF treatment. After 6 hours of treatment with HGF, occludin, claudin-1, and beta-catenin were redistributed from the membrane to the cytoplasm.

CONCLUSIONS

Treatment of RPE explants with HGF results in rapid disassembly of tight and adherens junctions associated with loss or redistribution of junctional proteins, decreased TER, and increased migration of RPE cells from the monolayer.

Authors+Show Affiliations

Department of Pathology, Keck School of Medicine of the University of Southern California Los Angeles, California 90033, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

12147616

Citation

Jin, Manlin, et al. "Regulation of RPE Intercellular Junction Integrity and Function By Hepatocyte Growth Factor." Investigative Ophthalmology & Visual Science, vol. 43, no. 8, 2002, pp. 2782-90.
Jin M, Barron E, He S, et al. Regulation of RPE intercellular junction integrity and function by hepatocyte growth factor. Invest Ophthalmol Vis Sci. 2002;43(8):2782-90.
Jin, M., Barron, E., He, S., Ryan, S. J., & Hinton, D. R. (2002). Regulation of RPE intercellular junction integrity and function by hepatocyte growth factor. Investigative Ophthalmology & Visual Science, 43(8), 2782-90.
Jin M, et al. Regulation of RPE Intercellular Junction Integrity and Function By Hepatocyte Growth Factor. Invest Ophthalmol Vis Sci. 2002;43(8):2782-90. PubMed PMID: 12147616.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Regulation of RPE intercellular junction integrity and function by hepatocyte growth factor. AU - Jin,Manlin, AU - Barron,Ernesto, AU - He,Shikun, AU - Ryan,Stephen J, AU - Hinton,David R, PY - 2002/7/31/pubmed PY - 2002/8/8/medline PY - 2002/7/31/entrez SP - 2782 EP - 90 JF - Investigative ophthalmology & visual science JO - Invest. Ophthalmol. Vis. Sci. VL - 43 IS - 8 N2 - PURPOSE: To evaluate the effect of hepatocyte growth factor (HGF) on the integrity and function of tight junctions and adherens junctions in the retinal pigment epithelial (RPE) monolayer. METHODS: Fresh bovine eyes were dissected to obtain 2- to 3-mm(2) explants of intact RPE with underlying choroid and sclera. Explants were cultured with or without HGF (20 ng/mL) for various periods (20 minutes to 72 hours). Junction integrity was assessed by transmission and scanning electron microscopy; localization, expression, and phosphorylation of junction proteins; and measurement of transepithelial resistance (TER), diffusion of fluorescent labeling in the plasma membrane, and the migration of RPE cells from the monolayer. RESULTS: Untreated explants consisted of polarized cells with apical microvilli and well-developed tight and adherens junctions. After HGF treatment, the explants showed loss of tight and adherens junctions ultrastructurally, diffusion of fluorescent label from apical to lateral membrane domains, and increased chemotactic migration of RPE cells from the monolayer. Primary cultures of confluent RPE cells showed a progressive decrease in TER. Western blot analysis showed rapid tyrosine phosphorylation of ZO-1, occludin, and beta-catenin within 20 minutes of stimulation. There was a marked loss of ZO-1 protein within 1 hour of HGF treatment. After 6 hours of treatment with HGF, occludin, claudin-1, and beta-catenin were redistributed from the membrane to the cytoplasm. CONCLUSIONS: Treatment of RPE explants with HGF results in rapid disassembly of tight and adherens junctions associated with loss or redistribution of junctional proteins, decreased TER, and increased migration of RPE cells from the monolayer. SN - 0146-0404 UR - https://www.unboundmedicine.com/medline/citation/12147616/Regulation_of_RPE_intercellular_junction_integrity_and_function_by_hepatocyte_growth_factor_ DB - PRIME DP - Unbound Medicine ER -