Nitric oxide and C-type atrial natriuretic peptide stimulate primary aortic smooth muscle cell migration via a cGMP-dependent mechanism: relationship to microfilament dissociation and altered cell morphology.
Migration of aortic smooth muscle cells is thought to be of essential importance in vascular restenosis, remodeling, and angiogenesis. Recent studies have shown that NO donors inhibit the migration of subcultured aortic smooth muscle cells. However, there is evidence that NO elicits opposite effects on cell proliferation in primary versus subcultured cells, indicating fundamental differences among different models of aortic smooth muscle cell cultures. The purpose of the current study was to investigate the effect of NO donors on migration of primary cultures of rat aortic smooth muscle cells and to compare and contrast their response with those in subcultured cells. A second purpose was to investigate some of the underlying mechanisms associated with NO-induced effects on cell migration. We report that 2 NO donors, S-nitroso-N-acetylpenicillamine (SNAP) and 2, 2-(hydroxynitrosohydrazino)bis-ethanamine, stimulated the migration of primary cells in a wounded-culture model as well as in a transwell migration model. The effect of NO donors was mimicked by 2 cGMP analogues and C-type natriuretic peptide and blocked by a specific inhibitor of guanyl cyclase, 1H-(1,2,4)oxadiazolo[4,3, -a]quinoxalin-1-one, indicating the involvement of cGMP as second messenger. Moreover, neither NO donors nor cGMP analogues altered migration of primary cultures stimulated by either FBS or angiotensin II. In contrast to its effect in primary cultures, SNAP did not alter basal or stimulated migration of subcultured cells, except at a relatively high concentration of 1 mmol/L, at which migration was inhibited. The migration-stimulatory effect of NO donors and cGMP was associated with altered cell morphology and dissociation of actin filaments, consistent with recent studies indicating that cell morphology and cytoskeletal organization influence cell migration. The results suggest the possible involvement of NO-induced cell migration in vascular injury or remodeling, representing conditions in which vascular NO levels would be expected to be elevated.
Brown
C
C
Department of Physiology and Biophysics, University of Tennessee, 894 Union Ave, Memphis, Tenn.38163-0001, USA. cbrown@physio1.utmem.edu
Pan
X
X
Hassid
A
A
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000378:metabolism*; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D006152:Cyclic GMP / Q000819:agonists / Q000502:physiology*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000096:biosynthesis; D003599:Cytoskeleton / Q000187:drug effects; D015194:Diffusion Chambers, Culture; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D018995:Myosin Heavy Chains / Q000096:biosynthesis; D020098:Natriuretic Peptide, C-Type / Q000502:physiology*; D009569:Nitric Oxide / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D010069:Oxadiazoles / Q000494:pharmacology; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D011810:Quinoxalines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley
C095284:1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one; D003470:Culture Media; D020030:Nitric Oxide Donors; D010069:Oxadiazoles; D011810:Quinoxalines; C110881:S-nitro-N-acetylpenicillamine; D000804:Angiotensin II; D020098:Natriuretic Peptide, C-Type; D015124:8-Bromo Cyclic Adenosine Monophosphate; D009569:Nitric Oxide; D000242:Cyclic AMP; D017869:Cyclic GMP-Dependent Protein Kinases; D018995:Myosin Heavy Chains; D006162:Guanylate Cyclase; D010396:Penicillamine; D006152:Cyclic GMP