Expression of the dystrophin-glycoprotein complex is a marker for human airway smooth muscle phenotype maturation.
Airway smooth muscle (ASM) cells may contribute to asthma pathogenesis through their capacity to switch between a synthetic/proliferative and a contractile phenotype. The multimeric dystrophin-glycoprotein complex (DGC) spans the sarcolemma, linking the actin cytoskeleton and extracellular matrix. The DGC is expressed in smooth muscle tissue, but its functional role is not fully established. We tested whether contractile phenotype maturation of human ASM is associated with accumulation of DGC proteins. We compared subconfluent, serum-fed cultures and confluent cultures subjected to serum deprivation, which express a contractile phenotype. Western blotting confirmed that beta-dystroglycan, beta-, delta-, and epsilon-sarcoglycan, and dystrophin abundance increased six- to eightfold in association with smooth muscle myosin heavy chain (smMHC) and calponin accumulation during 4-day serum deprivation. Immunocytochemistry showed that the accumulation of DGC subunits was specifically localized to a subset of cells that exhibit robust staining for smMHC. Laminin competing peptide (YIGSR, 1 microM) and phosphatidylinositol 3-kinase (PI3K) inhibitors (20 microM LY-294002 or 100 nM wortmannin) abrogated the accumulation of smMHC, calponin, and DGC proteins. These studies demonstrate that the accumulation of DGC is an integral feature for phenotype maturation of human ASM cells. This provides a strong rationale for future studies investigating the role of the DGC in ASM smooth muscle physiology in health and disease.
Sharma
Pawan
P
Department of Physiology, Section of Respiratory Disease, Univ. of Manitoba, Winnipeg, MB, Canada.
Tran
Thai
T
Stelmack
Gerald L
GL
McNeill
Karol
K
Gosens
Reinoud
R
Mutawe
Mark M
MM
Unruh
Helmut
H
Gerthoffer
William T
WT
Halayko
Andrew J
AJ
D002460:Cell Line; D016922:Cellular Senescence; D049030:Dystroglycans / Q000032:analysis; D016189:Dystrophin / Q000235:genetics*; D005819:Genetic Markers; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D009130:Muscle, Smooth / Q000166:cytology / Q000502:physiology*; D010641:Phenotype; D021122:Protein Subunits / Q000235:genetics; D012143:Respiratory Physiological Phenomena; D020133:Reverse Transcriptase Polymerase Chain Reaction; D049031:Sarcoglycans / Q000032:analysis; D019098:Telomerase / Q000032:analysis
D016189:Dystrophin; D005819:Genetic Markers; D006023:Glycoproteins; D021122:Protein Subunits; D049031:Sarcoglycans; D049030:Dystroglycans; D019098:Telomerase