skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Phenotypic heterogeneity influences the behavior of rat aortic smooth muscle cells in collagen lattice

Abstract

Phenotypic modulation of vascular smooth muscle cells (SMCs) in atherosclerosis and restenosis involves responses to the surrounding microenvironment. SMCs obtained by enzymatic digestion from tunica media of newborn, young adult (YA) and old rats and from the thickened intima (TI) and underlying media of young adult rat aortas 15 days after ballooning were entrapped in floating populated collagen lattice (PCL). TI-SMCs elongated but were poor at PCL contraction and remodeling and expressed less {alpha}2 integrin compared to other SMCs that appeared more dendritic. During early phases of PCL contraction, SMCs showed a marked decrease in the expression of {alpha}-smooth muscle actin and myosin. SMCs other than TI-SMCs required 7 days to re-express {alpha}-smooth muscle actin and myosin. Only TI-SMCs in PCL were able to divide in 48 h, with a greater proportion in S and G2-M cell cycle phases compared to other SMCs. Anti-{alpha}2 integrin antibody markedly inhibited contraction but not proliferation in YA-SMC-PLCs; anti-{alpha}1 and anti-{alpha}2 integrin antibodies induced a similar slight inhibition in TI-SMC-PCLs. Finally, TI-SMCs rapidly migrated from PCL on plastic reacquiring their epithelioid phenotype. Heterogeneity in proliferation and cytoskeleton as well the capacity to remodel the extracellular matrix are maintained, when SMCs are suspended in PCLs.

Authors:
 [1];  [2];  [3];  [2];  [4]
  1. Anatomic Pathology, Dept. of Biopathology and Image Diagnostics, Tor Vergata University of Rome, Via Montpellier 1, 00133 Rome (Italy). E-mail: orlandi@uniroma2.it
  2. Anatomic Pathology, Dept. of Biopathology and Image Diagnostics, Tor Vergata University of Rome, Via Montpellier 1, 00133 Rome (Italy)
  3. Department of Pathology and Immunology, University of Geneva (Switzerland)
  4. Division of Plastic Surgery, Hershey Medical Center, Hershey, PA (United States)
Publication Date:
OSTI Identifier:
20775322
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; Journal Volume: 311; Journal Issue: 2; Other Information: DOI: 10.1016/j.yexcr.2005.10.008; PII: S0014-4827(05)00476-3; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ACTIN; ANTIBODIES; AORTA; BALLOONING INSTABILITY; CATTLE; COLLAGEN; GROWTH FACTORS; INFANTS; MUSCLES; MYOSIN; RATS; TRANSMISSION ELECTRON MICROSCOPY

Citation Formats

Orlandi, Augusto, Ferlosio, Amedeo, Gabbiani, Giulio, Spagnoli, Luigi Giusto, and Ehrlich, Paul H. Phenotypic heterogeneity influences the behavior of rat aortic smooth muscle cells in collagen lattice. United States: N. p., 2005. Web. doi:10.1016/j.yexcr.2005.10.008.
Orlandi, Augusto, Ferlosio, Amedeo, Gabbiani, Giulio, Spagnoli, Luigi Giusto, & Ehrlich, Paul H. Phenotypic heterogeneity influences the behavior of rat aortic smooth muscle cells in collagen lattice. United States. doi:10.1016/j.yexcr.2005.10.008.
Orlandi, Augusto, Ferlosio, Amedeo, Gabbiani, Giulio, Spagnoli, Luigi Giusto, and Ehrlich, Paul H. Sat . "Phenotypic heterogeneity influences the behavior of rat aortic smooth muscle cells in collagen lattice". United States. doi:10.1016/j.yexcr.2005.10.008.
@article{osti_20775322,
title = {Phenotypic heterogeneity influences the behavior of rat aortic smooth muscle cells in collagen lattice},
author = {Orlandi, Augusto and Ferlosio, Amedeo and Gabbiani, Giulio and Spagnoli, Luigi Giusto and Ehrlich, Paul H.},
abstractNote = {Phenotypic modulation of vascular smooth muscle cells (SMCs) in atherosclerosis and restenosis involves responses to the surrounding microenvironment. SMCs obtained by enzymatic digestion from tunica media of newborn, young adult (YA) and old rats and from the thickened intima (TI) and underlying media of young adult rat aortas 15 days after ballooning were entrapped in floating populated collagen lattice (PCL). TI-SMCs elongated but were poor at PCL contraction and remodeling and expressed less {alpha}2 integrin compared to other SMCs that appeared more dendritic. During early phases of PCL contraction, SMCs showed a marked decrease in the expression of {alpha}-smooth muscle actin and myosin. SMCs other than TI-SMCs required 7 days to re-express {alpha}-smooth muscle actin and myosin. Only TI-SMCs in PCL were able to divide in 48 h, with a greater proportion in S and G2-M cell cycle phases compared to other SMCs. Anti-{alpha}2 integrin antibody markedly inhibited contraction but not proliferation in YA-SMC-PLCs; anti-{alpha}1 and anti-{alpha}2 integrin antibodies induced a similar slight inhibition in TI-SMC-PCLs. Finally, TI-SMCs rapidly migrated from PCL on plastic reacquiring their epithelioid phenotype. Heterogeneity in proliferation and cytoskeleton as well the capacity to remodel the extracellular matrix are maintained, when SMCs are suspended in PCLs.},
doi = {10.1016/j.yexcr.2005.10.008},
journal = {Experimental Cell Research},
number = 2,
volume = 311,
place = {United States},
year = {Sat Dec 10 00:00:00 EST 2005},
month = {Sat Dec 10 00:00:00 EST 2005}
}
  • OV-Phosphohydroxylysine was chemically synthesized and techniques were established for its identification by combined use of cation-exchange chromatography, thin-layer electrophoresis at pH 1.9 and 3.5, and thin-layer chromatography. Conditions were also determined to permit hydrolysis of proteins in 2 M HCl without loss of the phosphono group of phosphohydroxylysine residues. Experiments were then performed showing that TSP was incorporated into the hydroxylysine residues of cell-associated collagens when cultured calf aorta medial smooth muscle cells were incubated with (TSP)orthophosphate. In other experiments, the cells incorporated (TH)lysine into hydroxylysine residues of cell-associated collagen and then TSP into phosphohydroxylysine residues. The doubly labeled phosphohydroxylysinemore » subsequently isolated showed nearly 1:1 stoichiometry with respect to incorporation of precursor lysine and phosphorus. Finally, in preliminary experiments done with a cell-free extract of the smooth muscle cells, TSP was transferred from (el-TSP)ATP to hydroxylysine residues in several kinds of collagenous substrates. Thus, this work shows that smooth muscle cells have the capacity to phosphorylate hydroxylysine residues in their cell-associated collagens and provides preliminary evidence that a protein kinase is involved.« less
  • In the presence of endothelin, there was a rapid increase in the /sup 45/Ca++ efflux from primary cultured rat vascular smooth muscle cells, both in physiological salt solution and in calcium free medium containing 2 mM EGTA. The /sup 45/Ca++ influx was not affected. The endothelin-induced, transient increase in cytosolic calcium concentration is probably mainly due to release of calcium from the intracellular store in vascular smooth muscle cells.
  • Cultured arterial smooth muscle cells (SMC) can produce platelet-derived growth factor (PDGF)-like molecules. This property raises the possibility that SMC-derived PDGFs function as autocrine/paracrine regulators in the formation and maintenance of the artery wall. In this study the authors have asked if levels of mRNAs directing synthesis of PDFG are modulated in aortic SMC during postnatal development. The authors report here that genes encoding PDGF A- and B-chain precursors are expressed at similar low levels in intact aortas from newborn and adult rats. Marked differences in regulation of transcript abundance of these genes were revealed when aortic SMC were grownmore » in cell culture. PDGF B-chain transcripts accumulated in passaged newborn rat SMC but not adult rat SMC, whereas PDGF A-chain RNA was found in comparable amounts in SMC from both age groups. Similarly, SMC from newborn rats secreted at least 60-fold more PDGF-like activity into conditioned medium than did adult rat SMC. These results show that PDGF A- and B-chain genes are transcribed in the normal rat aorta and provide evidence for age-related change in the control of PDGF B-chain gene expression in aortic SMC. Independent regulation of transcript levels in cultured SMC leaves open the possibility that PDGFs of different composition (AA, AB, BB) play different roles in normal function of the artery wall.« less
  • The influence of cell density on the binding characteristics of thromboxane A{sub 2}/prostaglandin H{sub 2} (TXA{sub 2}/PGH{sub 2}) receptors in rat aortic vascular smooth muscle cells in culture were determined using (1S- (1{alpha}, 2{beta} (5Z), 3a (1E, 3R*), 4{alpha})) - (3- (3-hydroxy-4- (4{prime}-iodophenoxy)-1-butyenyl)-7-oxabicyclo-(2.2.1)heptan-2yl)-5-heptenoic acid ({sup 125}I-BOP). The B{sub max} for {sup 125}I-BOP was 5,430 {plus minus} 139 sites/cell (26.9 {plus minus} 5.7 fmoles/mg protein) for cells cultured in 1% fetal calf serum and 2,809 {plus minus} 830 sites/cell (13.1 {plus minus} 2.2 fmoles/mg protein) for cells cultured in 10% fetal calf serum. Cells were allowed to grow to varying densitiesmore » and then harvested for assay. There was a negative correlation between the B{sub max} and the cell density per flask. The K{sub d} for I-BOP did not significantly vary in any of the studies. The results demonstrate that cell density plays an important role in influencing the expression of vascular TXA{sub 2}/PGH{sub 2} receptors.« less
  • Short communication.