It is well documented that cell-matrix interactions play an important regulatory role in controlling cell behavior (growth, differentiation, gene expression, and organization of cytoskeleton) and the organization of the extracellular matrix. The importance of cell-matrix interactions on the trabecular meshwork (TM) and aqueous outflow, however, is not known. Preliminary studies suggest that integrin/syndecan mediated signaling events could control aqueous outflow, since the heparin II (Hep II) domain of fibronectin that contains alpha4 beta1 integrin and syndecan binding sites lowers intraocular pressure in human eye organ cultures. The goal of this grant is to determine (1) whether outflow facility is increased because the Hep II domain affects the formation of cell contacts (cell-cell and cell-matrix) interactions and /or the organization of the extracellular matrix in the TM and (2) if the Hep II domain uses interactions with alpha4 beta1 integrins and syndecans to modulate outflow facility. The effect of the Hep II domain of fibronectin on aqueous outflow will be determined by perfusing the anterior chamber of primates or human eye organ cultures with recombinant Hep II domains and then analyzing its effects on aqueous outflow and the ultrastructure of the TM (cell contacts and/or matrix formation). Recombinant Hep II domains containing mutations in the alpha4 beta1 and /or syndecan binding sites will be used to determine the role of integrins and syndecans in aqueous outflow. The effect of the Hep II domain on cell contacts and the formation of matrix will be done by using various biochemical (zymograms, immunoprecipitation, Northern and Western blots) and microscopy (immunofluorescence, electron, and in situ hybridization) procedures to determine the effect of the Hep II domain on the localization of proteins in cell contacts (FAK, paxillin, cadherins, catenins and vinculin) and the expression of matrix proteins (stromelysin, gelatinases, fibronectin, laminin, and collagen). These studies should demonstrate how matrix-mediated interactions may influence aqueous outflow. Ultimately, such information should prove useful in the development of therapeutic peptides to control glaucoma.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
1R01EY012515-01A2
Application #
6326558
Study Section
Visual Sciences A Study Section (VISA)
Program Officer
Liberman, Ellen S
Project Start
2001-05-01
Project End
2006-04-30
Budget Start
2001-05-01
Budget End
2002-04-30
Support Year
1
Fiscal Year
2001
Total Cost
$254,625
Indirect Cost
Name
University of Wisconsin Madison
Department
Pathology
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Faralli, Jennifer A; Newman, Jessica R; Sheibani, Nader et al. (2011) Integrin-linked kinase regulates integrin signaling in human trabecular meshwork cells. Invest Ophthalmol Vis Sci 52:1684-92
Schwinn, Marie K; Gonzalez Jr, Jose M; Gabelt, B'Ann T et al. (2010) Heparin II domain of fibronectin mediates contractility through an alpha4beta1 co-signaling pathway. Exp Cell Res 316:1500-12
Tan, James C H; Kiland, Julie A; Gonzalez, Jose M et al. (2010) Sodium orthovanadate effect on outflow facility and intraocular pressure in live monkeys. Exp Eye Res 91:486-90
Faralli, Jennifer A; Schwinn, Marie K; Gonzalez Jr, Jose M et al. (2009) Functional properties of fibronectin in the trabecular meshwork. Exp Eye Res 88:689-93
Filla, Mark S; Schwinn, Marie K; Sheibani, Nader et al. (2009) Regulation of cross-linked actin network (CLAN) formation in human trabecular meshwork (HTM) cells by convergence of distinct beta1 and beta3 integrin pathways. Invest Ophthalmol Vis Sci 50:5723-31
Gonzalez Jr, Jose M; Hu, Yujie; Gabelt, B'Ann T et al. (2009) Identification of the active site in the heparin II domain of fibronectin that increases outflow facility in cultured monkey anterior segments. Invest Ophthalmol Vis Sci 50:235-41
Maqueda, Alfredo; Moyano, Jose V; Hernandez Del Cerro, Mercedes et al. (2007) The heparin III-binding domain of fibronectin (III4-5 repeats) binds to fibronectin and inhibits fibronectin matrix assembly. Matrix Biol 26:642-51
Tan, James C H; Peters, Donna M; Kaufman, Paul L (2006) Recent developments in understanding the pathophysiology of elevated intraocular pressure. Curr Opin Ophthalmol 17:168-74
Filla, Mark S; Woods, Anne; Kaufman, Paul L et al. (2006) Beta1 and beta3 integrins cooperate to induce syndecan-4-containing cross-linked actin networks in human trabecular meshwork cells. Invest Ophthalmol Vis Sci 47:1956-67
Gonzalez Jr, Jose M; Faralli, Jennifer A; Peters, Joanne M et al. (2006) Effect of heparin II domain of fibronectin on actin cytoskeleton and adherens junctions in human trabecular meshwork cells. Invest Ophthalmol Vis Sci 47:2924-31

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