Differential effects of laminin, intact type IV collagen, and specific domains of type IV collagen on endothelial cell adhesion and migration.

Abstract

Abstract. Laminin and type IV collagen were compared for the ability to promote aortic endothelial cell adhesion and directed migration in vitro. Substratumadsorbed IV promoted aortic endothelial cell adhesion in a concentration dependent fashion attaining a maximum level 141-fold greater than controls within 30 min. Aortic endothelial cell adhesion to type IV collagen was not inhibited by high levels (10 -3 M) of arginyl-glycyl-aspartyl-serine. In contrast, adhesion of aortic endothelial cells on laminin was slower, attaining only 53 % of the adhesion observed on type IV collagen by 90 min. Type IV collagen when added to the lower well of a Boyden chamber stimulated the directional migration of aortic endothelial cells in a concentration dependent manner with a maximal response 6.9-fold over control levels, whereas aortic endothelial cells did not migrate in response to laminin at any concentration (.01-2.0 x 10 -7 M). Triple helix-rich fragments of type IV collagen were nearly as active as intact type IV collagen in stimulating both adhesion and migration whereas the carboxy terminal globular domain was less active at promoting adhesion (36% of the adhesion promoted by intact type IV collagen) or migration. Importantly, aortic endothelial cells also migrate to substratum adsorbed gradients of type IV collagen suggesting that the mechanism of migration is haptotactic in nature. These results demonstrate that the aortic endothelial cell adhesion and migration is preferentially promoted by type IV collagen compared with laminin, and has a complex molecular basis which may be important in angiogenesis and large vessel repair.

Mammalian collagen IV

“…In addition to the triple-helical part, NC1 domain of collagen IV also promotes cell adhesion (Chelberg et al, 1989;Herbst et al, 1988;Setty et al, 1998), stimulates outgrowth of embryonic neurons (Lein et al, 1991), and inhibits morphogenesis in Hydra vulgaris (Zhang et al, 1994). The latter finding led to the suggestion that NC1 domain could inhibit angiogenesis by disrupting the assembly of vascular basement membrane (Sarras and Hudson, 1997).…”

“…The classical example of integrin binding site is the Arg-Gly-Asp (RGD) sequence originally identified in fibronectin, which is a common ligand for at least eight different integrins (Ruoslahti, 1996). Although multiple RGD sequences exist within the triple-helical region of several collagen IV α-chains, several studies have shown that cell adhesion to collagen IV is RGD-independent (Herbst et al, 1988;Kim et al, 1994;Kramer and Marks, 1989). A possible explanation is that due to their triple-helical nature, these RGD sequences are not accessible to integrins.…”

“…This interaction is critical for a variety of biological processes, including cell adhesion, migration, survival, proliferation, and differentiation. Cell culture studies have shown that collagen IV is the binding substrate for a large number of cell types including platelets (Santoro, 1986;Staatz et al, 1990), hepatocytes (Rubin et al, 1981), keratinocytes (Murray et al, 1979), endothelial (Cheng and Kramer, 1989;Herbst et al, 1988), mesangial (Setty et al, 1998), pancreatic (Kaido et al, 2004), and tumor cells such as breast and prostate carcinoma (Abecassis et al, 1987;Dedhar et al, 1993), melanoma (Chelberg et al, 1989), fibrosarcoma, and glioma (Aumailley and Timpl, 1986;Knight et al, 2000). Cell attachment to collagen IV is mediated by multiple binding sites within both triple-helical and NC1 domains, suggesting involvement of several adhesion receptors (Chelberg et al, 1989;Herbst et al, 1988;Wayner and Carter, 1987).…”

COL4A2 mutation associated with familial porencephaly and small-vessel disease

“…28 The non-collagen domain of collagen IV alpha-2, but not alpha-1 chain, promotes cell adhesion, outgrowth of embryonic neurons and is responsible for strict regulation of the 2:1 stoichiometry of the triple helix. 18,[29][30][31] This could indicate that a disturbed ratio of procollagen alpha-1 and procollagen alpha-2 chains due to a heterozygous null allele of either COL4A1 or COL4A2 may lead to a disturbed or insufficient heterotrimer assembly and secretion.…”

Decellularization of human donor aortic and pulmonary valved conduits using low concentration sodium dodecyl sulfate

“…This included analysis for collagen IV, which was used as a marker for the basement membrane, which is believed to be important for endothelialization (Herbst et al ., 1988) and VWF, which could potentially have the undesirable effect of increasing platelet adhesion to the decellularized matrix if in direct blood contact. Both collagen IV and VWF were present in the decellularized tissues.…”