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Galectin-1 influences migration of retinal pigment epithelial cells.
Invest Ophthalmol Vis Sci. 2006 Jan; 47(1):415-26.IO

Abstract

PURPOSE

To determine whether the beta-galactoside-binding matricellular protein Gal-1 is expressed in human specimens of proliferative vitreoretinopathy (PVR) and to evaluate its influence on RPE migration.

METHODS

RT-PCR was used to detect Gal-1-specific transcripts in PVR membranes, and the expression pattern of Gal-1 was examined by immunohistochemistry. Expression of Gal-1 in native, low- and high-density cultured RPE cells was determined by Western blot analysis. Cultured human RPE cells were treated with bFGF, TGF-beta2, PDGF-BB, or HGF. The dose-response of Gal-1 mRNA expression was measured by by real-time quantitative RT-PCR and Northern blot analysis. Induction of Gal-1 protein was confirmed by Western blot analysis. To study the effect of Gal-1 on RPE migration in vitro, Gal-1 expression was silenced by RNA interference. beta-Lactose was used to saturate extracellular galectins. RPE cell migration was assessed by a modified Boyden chamber assay, with HGF as the chemoattractant.

RESULTS

Gal-1 mRNA expression was present in human specimens of PVR membranes, and staining for Gal-1 was distributed throughout the extracellular matrix (ECM) of PVR membranes. Colocalization was found with laminin and fibronectin and cells of epithelial origin. Western blot analysis revealed greater baseline expression levels in low-density cultured RPE cells than in native and high-density cultured RPE cells. Treatment with HGF caused a dose-dependent increase in Gal-1 expression. Low expression levels of Gal-1 correlated with a reduction of RPE migration to 14% of control. beta-Lactose inhibited HGF-induced RPE cell migration to 23% of control.

CONCLUSIONS

Gal-1 is present in the extracellular matrix of PVR membranes and may be derived from dedifferentiated RPE cells. The expression level of Gal-1 appears to be related to a migratory RPE phenotype and stimulation by HGF, both conditions implicated in the pathogenesis of early PVR. Furthermore, HGF-induced RPE migration may be dependent, at least in part, on Gal-1- and beta-galactoside-dependent mechanisms.

Authors+Show Affiliations

Department of Ophthalmology, Ludwig-maximilians-University, Munich, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

16384992

Citation

Alge, Claudia S., et al. "Galectin-1 Influences Migration of Retinal Pigment Epithelial Cells." Investigative Ophthalmology & Visual Science, vol. 47, no. 1, 2006, pp. 415-26.
Alge CS, Priglinger SG, Kook D, et al. Galectin-1 influences migration of retinal pigment epithelial cells. Invest Ophthalmol Vis Sci. 2006;47(1):415-26.
Alge, C. S., Priglinger, S. G., Kook, D., Schmid, H., Haritoglou, C., Welge-Lussen, U., & Kampik, A. (2006). Galectin-1 influences migration of retinal pigment epithelial cells. Investigative Ophthalmology & Visual Science, 47(1), 415-26.
Alge CS, et al. Galectin-1 Influences Migration of Retinal Pigment Epithelial Cells. Invest Ophthalmol Vis Sci. 2006;47(1):415-26. PubMed PMID: 16384992.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Galectin-1 influences migration of retinal pigment epithelial cells. AU - Alge,Claudia S, AU - Priglinger,Siegfried G, AU - Kook,Daniel, AU - Schmid,Holger, AU - Haritoglou,Christos, AU - Welge-Lussen,Ulrich, AU - Kampik,Anselm, PY - 2005/12/31/pubmed PY - 2006/2/10/medline PY - 2005/12/31/entrez SP - 415 EP - 26 JF - Investigative ophthalmology & visual science JO - Invest. Ophthalmol. Vis. Sci. VL - 47 IS - 1 N2 - PURPOSE: To determine whether the beta-galactoside-binding matricellular protein Gal-1 is expressed in human specimens of proliferative vitreoretinopathy (PVR) and to evaluate its influence on RPE migration. METHODS: RT-PCR was used to detect Gal-1-specific transcripts in PVR membranes, and the expression pattern of Gal-1 was examined by immunohistochemistry. Expression of Gal-1 in native, low- and high-density cultured RPE cells was determined by Western blot analysis. Cultured human RPE cells were treated with bFGF, TGF-beta2, PDGF-BB, or HGF. The dose-response of Gal-1 mRNA expression was measured by by real-time quantitative RT-PCR and Northern blot analysis. Induction of Gal-1 protein was confirmed by Western blot analysis. To study the effect of Gal-1 on RPE migration in vitro, Gal-1 expression was silenced by RNA interference. beta-Lactose was used to saturate extracellular galectins. RPE cell migration was assessed by a modified Boyden chamber assay, with HGF as the chemoattractant. RESULTS: Gal-1 mRNA expression was present in human specimens of PVR membranes, and staining for Gal-1 was distributed throughout the extracellular matrix (ECM) of PVR membranes. Colocalization was found with laminin and fibronectin and cells of epithelial origin. Western blot analysis revealed greater baseline expression levels in low-density cultured RPE cells than in native and high-density cultured RPE cells. Treatment with HGF caused a dose-dependent increase in Gal-1 expression. Low expression levels of Gal-1 correlated with a reduction of RPE migration to 14% of control. beta-Lactose inhibited HGF-induced RPE cell migration to 23% of control. CONCLUSIONS: Gal-1 is present in the extracellular matrix of PVR membranes and may be derived from dedifferentiated RPE cells. The expression level of Gal-1 appears to be related to a migratory RPE phenotype and stimulation by HGF, both conditions implicated in the pathogenesis of early PVR. Furthermore, HGF-induced RPE migration may be dependent, at least in part, on Gal-1- and beta-galactoside-dependent mechanisms. SN - 0146-0404 UR - https://www.unboundmedicine.com/medline/citation/16384992/Galectin_1_influences_migration_of_retinal_pigment_epithelial_cells_ L2 - http://iovs.arvojournals.org/article.aspx?doi=10.1167/iovs.05-0308 DB - PRIME DP - Unbound Medicine ER -