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Type IV collagenase(s) and TIMPs modulate endothelial cell morphogenesis in vitro
H W Schnaper1, D S Grant, W G Stetler-Stevenson
1Laboratory of Developmental Biology, National Institute of Dental Research, Bethesda, Maryland 20892.
Journal of Cellular Physiology
|August 1, 1993
Summary
Protease activity, specifically gelatinase/type IV collagenase, is crucial for endothelial cell tube formation on Matrigel. Inhibiting this activity or adding more enzyme significantly impacts capillary-like network development.
Area of Science:
- Cell Biology
- Biochemistry
- Angiogenesis Research
Background:
- Proteases are implicated in endothelial cell functions.
- The gelatinase/type IV collagenase system's role in endothelial differentiation requires further investigation.
Purpose of the Study:
- To investigate the contribution of the gelatinase/type IV collagenase system to endothelial cell differentiation in vitro.
- To elucidate the role of specific gelatinases and their inhibitors in endothelial morphogenesis.
Main Methods:
- Utilized an in vitro model of endothelial differentiation using human umbilical vein endothelial cells cultured on Matrigel.
- Analyzed gelatinase activity via zymography.
- Assessed the impact of antibodies against type IV collagenase and tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) on tube formation.
- Investigated the effect of exogenous recombinant 72-kD gelatinase and measured mRNA expression levels.
Main Results:
- Endothelial cells cultured on Matrigel formed tube networks, with detected 72-kD (cell-derived) and 92-kD (Matrigel-derived) gelatinase activities.
- Antibodies against type IV collagenase and TIMPs significantly reduced tube network area.
- Exogenous 72-kD gelatinase enhanced tube formation.
- Inhibition was most effective when applied early in culture, indicating a role in initial morphogenesis.
- mRNA expression for 72-kD gelatinase decreased during culture, while TIMP mRNA increased.
Conclusions:
- Gelatinase/type IV collagenase activity is essential for endothelial cell morphogenesis on Matrigel.
- Collagenases likely facilitate an early event in endothelial tube formation.
- These findings suggest a role for collagenases in in vivo capillary formation (angiogenesis).