Extracellular matrix molecules regulate endothelial cell migration stimulated by lysophosphatidic acid.
BACKGROUND
Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) are lipids that bind G-protein coupled receptors and differentially promote transmigration of endothelial cells.


OBJECTIVE
To determine if endothelial cell transmigration stimulated by LPA, not S1P, is dependent on the extracellular matrix.


METHODS
Bovine pulmonary artery (BPAE) endothelial cell transmigration and locomotion were measured using a modified-Boyden chamber and video microscopy, respectively. Results were related to strength of adhesion and characteristics of cell adhesive contacts.


CONCLUSIONS
BPAEs responded to LPA by transmigration through gelatin- or collagen-coated filters, but not through fibronectin-, vitronectin-, or fibrinogen-coated filters. Fewer cells adhered to collagen or gelatin than to fibronectin in a static cell adhesion assay or after application of a g-force to detach cells. Video microscopy revealed that S1P stimulates large lamellipodia on two-dimensional fibronectin substrate. LPA stimulated lamellipodia on fibronectin, but the trailing edge remained attached, resulting in sting ray-shaped cells in video microscopy. LPA-treated cells on gelatin released the trailing edge. To understand how the extracellular matrix may regulate endothelial cell shape during movement, we surveyed changes in focal adhesion proteins. More Hic-5, a paxillin homolog, was detected in the detergent insoluble fraction of BPAEs attached to gelatin than fibronectin. No such difference was found in paxillin. In BPAEs, Hic-5 was localized to smaller punctate structures on fibronectin and longer, thinner focal adhesions on gelatin. These results indicated that localization of Hic-5 and strength of adhesion correlate with endothelial cell transmigration stimulated by LPA, but not with transmigration stimulated by S1P.
Panetti
T S
TS
Department of Microbiology and Immunology, Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA 19140, USA. tpanetti@temple.edu
Hannah
D F
DF
Avraamides
C
C
Gaughan
J P
JP
Marcinkiewicz
C
C
Huttenlocher
A
A
Mosher
D F
DF
D000818:Animals; D002417:Cattle; D002448:Cell Adhesion; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D019483:Disintegrins / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D008246:Lysophospholipids / Q000494:pharmacology*; D018715:Microscopy, Video; D009842:Oligopeptides / Q000494:pharmacology; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology
D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D019483:Disintegrins; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D008246:Lysophospholipids; D009842:Oligopeptides; D051419:Paxillin; D010750:Phosphoproteins; C490351:TGFB1I1 protein, human; C060506:sphingosine 1-phosphate; C047981:arginyl-glycyl-aspartic acid; D005780:Gelatin; D013110:Sphingosine; C032881:lysophosphatidic acid