Type IV collagenase(s) and TIMPs modulate endothelial cell morphogenesis in vitro

H. William Schnaper, Derrick S. Grant, William G. Stetler-Stevenson, Rafael Fridman, Gabriella D'Orazi, Anne N. Murphy, Robert E. Bird, Matti Hoythya, Thomas R. Fuerst, Deborah L. French, James P. Quigley, Hynda K. Kleinman

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Abstract

It has been proposed that proteases are important in endothelial cell behavior. We examined the contribution of the gelatinase/type IV collagenase system in an in vitro model of endothelial differentiation. Human umbilical vein endothelial cells rapidly align and form networks of tubes when cultured on a basement membrane preparation, Matrigel. Zymograms of culture supernates demonstrate a 72-kD and a 92-kD gelatinase activity; the cells produce most of the 72-kD gelatinase, whereas the 92-kD activity is derived entirely from the Matrigel. Addition of antibodies against type IV gelatinase/collagenase decreases the area of the tube network. Both tissue inhibitors of metalloproteinases, TIMP-1 and TIMP-2, similarly decrease tube formation when added to cultures. Conversely, exogenous recombinant 72-kD gelatinase increases tube-forming activity. The effects of the anti-gelatinase antibodies and the TIMPs are not additive. Inhibition by either antibodies or TIMPs is greatest when they are added at culture initiation, suggesting that the protease activity is important in the early steps of morphogenesis. However, culture of the cells on Matrigel does not increase early expression of mRNA for the 72-kD gelatinase. Expression of message for the enzyme actually decreases during the course of the assay, while transcription of mRNAs for TIMPs increases, further supporting the concept that collagenases facilitate an early event in tube formation. These data demonstrate that gelatinase/type IV collagenase activity is important in endothelial cell morphogenesis on Matrigel, and suggest a role for collagenases in formation of new capillaries in vivo.

Original languageEnglish
Pages (from-to)235-246
Number of pages12
JournalJournal of Cellular Physiology
Volume156
Issue number2
Publication statusPublished - Aug 1993

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Gelatinases
Endothelial cells
Collagenases
Morphogenesis
Endothelial Cells
Tissue Inhibitor of Metalloproteinase-1
Antibodies
Peptide Hydrolases
Tissue Inhibitor of Metalloproteinases
Tissue Inhibitor of Metalloproteinase-2
Messenger RNA
In Vitro Techniques
Human Umbilical Vein Endothelial Cells
Transcription
Basement Membrane
Anti-Idiotypic Antibodies
Assays
Cell Culture Techniques
matrigel
Enzymes

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

Cite this

Schnaper, H. W., Grant, D. S., Stetler-Stevenson, W. G., Fridman, R., D'Orazi, G., Murphy, A. N., ... Kleinman, H. K. (1993). Type IV collagenase(s) and TIMPs modulate endothelial cell morphogenesis in vitro. Journal of Cellular Physiology, 156(2), 235-246.

Type IV collagenase(s) and TIMPs modulate endothelial cell morphogenesis in vitro. / Schnaper, H. William; Grant, Derrick S.; Stetler-Stevenson, William G.; Fridman, Rafael; D'Orazi, Gabriella; Murphy, Anne N.; Bird, Robert E.; Hoythya, Matti; Fuerst, Thomas R.; French, Deborah L.; Quigley, James P.; Kleinman, Hynda K.

In: Journal of Cellular Physiology, Vol. 156, No. 2, 08.1993, p. 235-246.

Research output: Contribution to journalArticle

Schnaper, HW, Grant, DS, Stetler-Stevenson, WG, Fridman, R, D'Orazi, G, Murphy, AN, Bird, RE, Hoythya, M, Fuerst, TR, French, DL, Quigley, JP & Kleinman, HK 1993, 'Type IV collagenase(s) and TIMPs modulate endothelial cell morphogenesis in vitro', Journal of Cellular Physiology, vol. 156, no. 2, pp. 235-246.
Schnaper HW, Grant DS, Stetler-Stevenson WG, Fridman R, D'Orazi G, Murphy AN et al. Type IV collagenase(s) and TIMPs modulate endothelial cell morphogenesis in vitro. Journal of Cellular Physiology. 1993 Aug;156(2):235-246.
Schnaper, H. William ; Grant, Derrick S. ; Stetler-Stevenson, William G. ; Fridman, Rafael ; D'Orazi, Gabriella ; Murphy, Anne N. ; Bird, Robert E. ; Hoythya, Matti ; Fuerst, Thomas R. ; French, Deborah L. ; Quigley, James P. ; Kleinman, Hynda K. / Type IV collagenase(s) and TIMPs modulate endothelial cell morphogenesis in vitro. In: Journal of Cellular Physiology. 1993 ; Vol. 156, No. 2. pp. 235-246.
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