Limits...
Localization of TGF-beta type II receptor and ED-A fibronectin in normal conjunctiva and failed filtering blebs.

Meyer-Ter-Vehn T, Grehn F, Schlunck G - Mol. Vis. (2008)

Bottom Line: TGF-beta receptor type II (TGF-beta-RII), alpha-smooth muscle actin, O-linked sialoglycoprotein, fibronectin and the ED-A fibronectin splice-variant were also detected using specific antibodies.Labeled sections were viewed with a confocal laser scanning microscope.Vascular endothelial cells expressed TGF-beta-RII in both normal and scarred tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of Würzburg, Würzburg, Germany. meyer_T3@augenklinik.uni-wuerzburg.de

ABSTRACT

Purpose: The cytokine transforming growth factor-beta (TGF-beta), and the ED-A splice variant of the extracellular matrix protein fibronectin modulate wound healing and scar formation. To further elucidate their possible role in filtering bleb scarring after glaucoma surgery in human eyes in vivo, we studied the cell type specific localization of TGF-beta receptors and the presence of ED-A fibronectin in sections of normal conjunctiva and scarred filtering blebs.

Methods: Cryosections of normal conjunctiva (four patients) and scarred filtering blebs (seven patients) were studied by double-label immunofluorescence. Antibodies against PECAM-1 and prolyl-4-hydroxylase allowed for specification of vascular endothelial cells and activated fibroblasts, respectively. TGF-beta receptor type II (TGF-beta-RII), alpha-smooth muscle actin, O-linked sialoglycoprotein, fibronectin and the ED-A fibronectin splice-variant were also detected using specific antibodies. Labeled sections were viewed with a confocal laser scanning microscope.

Results: Vascular endothelial cells expressed TGF-beta-RII in both normal and scarred tissue. TGF-beta-RII was sparsely detected in the fibroblasts of normal conjunctiva while it was strongly expressed in most fibroblasts of the scarred filtering blebs. Similarly, ED-A fibronectin was not detected in the extracellular matrix of normal conjunctiva but abundantly present in scarred filtering blebs.

Conclusions: Filtering bleb scarring is associated with an abundant expression of TGF-beta receptors in activated fibroblasts and the deposition of the fibrogenic ED-A fibronectin splice-variant. These data support the concept of targeting TGF-beta signaling to prevent scar formation after filtering glaucoma surgery.

Show MeSH

Related in: MedlinePlus

Expression of fibronectin and the ED-A fibronectin splice variant. Fibronectin is abundant in normal conjunctiva (A) and in scarred filtering blebs (B). In contrast, expression of the fibrogenic ED-A fibronectin splice variant is restricted to perivascular areas in normal conjunctiva (A), but expands throughout the tissue in scarred filtering blebs (B). Specimens were double-stained for total fibronectin (green) and the fibronectin isoform, ED-A (red). Scale bar represents 50 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2255025&req=5

f5: Expression of fibronectin and the ED-A fibronectin splice variant. Fibronectin is abundant in normal conjunctiva (A) and in scarred filtering blebs (B). In contrast, expression of the fibrogenic ED-A fibronectin splice variant is restricted to perivascular areas in normal conjunctiva (A), but expands throughout the tissue in scarred filtering blebs (B). Specimens were double-stained for total fibronectin (green) and the fibronectin isoform, ED-A (red). Scale bar represents 50 µm.

Mentions: To study fibronectin extracellular matrix composition in conjunctival tissue, we performed double stains using antibodies against total fibronectin and the ED-A fibronectin splice variant. Fibronectin was found throughout the subepithelial stroma in both the normal conjunctiva and scarred filtering blebs (Figure 5A,B). In normal conjunctiva, expression of the ED-A fibronectin splice variant was restricted to perivascular areas and basal epithelial cells (Figure 5A). In contrast, ED-A fibronectin was present throughout the stroma of scarred filtering blebs (Figure 5B).


Localization of TGF-beta type II receptor and ED-A fibronectin in normal conjunctiva and failed filtering blebs.

Meyer-Ter-Vehn T, Grehn F, Schlunck G - Mol. Vis. (2008)

Expression of fibronectin and the ED-A fibronectin splice variant. Fibronectin is abundant in normal conjunctiva (A) and in scarred filtering blebs (B). In contrast, expression of the fibrogenic ED-A fibronectin splice variant is restricted to perivascular areas in normal conjunctiva (A), but expands throughout the tissue in scarred filtering blebs (B). Specimens were double-stained for total fibronectin (green) and the fibronectin isoform, ED-A (red). Scale bar represents 50 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2255025&req=5

f5: Expression of fibronectin and the ED-A fibronectin splice variant. Fibronectin is abundant in normal conjunctiva (A) and in scarred filtering blebs (B). In contrast, expression of the fibrogenic ED-A fibronectin splice variant is restricted to perivascular areas in normal conjunctiva (A), but expands throughout the tissue in scarred filtering blebs (B). Specimens were double-stained for total fibronectin (green) and the fibronectin isoform, ED-A (red). Scale bar represents 50 µm.
Mentions: To study fibronectin extracellular matrix composition in conjunctival tissue, we performed double stains using antibodies against total fibronectin and the ED-A fibronectin splice variant. Fibronectin was found throughout the subepithelial stroma in both the normal conjunctiva and scarred filtering blebs (Figure 5A,B). In normal conjunctiva, expression of the ED-A fibronectin splice variant was restricted to perivascular areas and basal epithelial cells (Figure 5A). In contrast, ED-A fibronectin was present throughout the stroma of scarred filtering blebs (Figure 5B).

Bottom Line: TGF-beta receptor type II (TGF-beta-RII), alpha-smooth muscle actin, O-linked sialoglycoprotein, fibronectin and the ED-A fibronectin splice-variant were also detected using specific antibodies.Labeled sections were viewed with a confocal laser scanning microscope.Vascular endothelial cells expressed TGF-beta-RII in both normal and scarred tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, University of Würzburg, Würzburg, Germany. meyer_T3@augenklinik.uni-wuerzburg.de

ABSTRACT

Purpose: The cytokine transforming growth factor-beta (TGF-beta), and the ED-A splice variant of the extracellular matrix protein fibronectin modulate wound healing and scar formation. To further elucidate their possible role in filtering bleb scarring after glaucoma surgery in human eyes in vivo, we studied the cell type specific localization of TGF-beta receptors and the presence of ED-A fibronectin in sections of normal conjunctiva and scarred filtering blebs.

Methods: Cryosections of normal conjunctiva (four patients) and scarred filtering blebs (seven patients) were studied by double-label immunofluorescence. Antibodies against PECAM-1 and prolyl-4-hydroxylase allowed for specification of vascular endothelial cells and activated fibroblasts, respectively. TGF-beta receptor type II (TGF-beta-RII), alpha-smooth muscle actin, O-linked sialoglycoprotein, fibronectin and the ED-A fibronectin splice-variant were also detected using specific antibodies. Labeled sections were viewed with a confocal laser scanning microscope.

Results: Vascular endothelial cells expressed TGF-beta-RII in both normal and scarred tissue. TGF-beta-RII was sparsely detected in the fibroblasts of normal conjunctiva while it was strongly expressed in most fibroblasts of the scarred filtering blebs. Similarly, ED-A fibronectin was not detected in the extracellular matrix of normal conjunctiva but abundantly present in scarred filtering blebs.

Conclusions: Filtering bleb scarring is associated with an abundant expression of TGF-beta receptors in activated fibroblasts and the deposition of the fibrogenic ED-A fibronectin splice-variant. These data support the concept of targeting TGF-beta signaling to prevent scar formation after filtering glaucoma surgery.

Show MeSH
Related in: MedlinePlus