Rho-kinase mediated cytoskeletal stiffness in skinned smooth muscle.

TitleRho-kinase mediated cytoskeletal stiffness in skinned smooth muscle.
Publication TypeJournal Article
Year of Publication2013
AuthorsLan, B, Wang, L, Zhang, J, Pascoe, CD, Norris, BA, Liu, JC-Y, Solomon, D, Paré, PD, Deng, L, Seow, CY
JournalJ Appl Physiol (1985)
Volume115
Issue10
Pagination1540-52
Date Published2013 Nov
ISSN1522-1601
Keywords1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine, Acetylcholine, Animals, Calcium, Cell Shape, Cytoskeleton, Elasticity, Muscle Contraction, Myocytes, Smooth Muscle, Myosin Light Chains, Phosphorylation, Protein Kinase Inhibitors, rho-Associated Kinases, Sheep, Signal Transduction, Time Factors, Trachea
Abstract

The structurally dynamic cytoskeleton is important in many cell functions. Large gaps still exist in our knowledge regarding what regulates cytoskeletal dynamics and what underlies the structural plasticity. Because Rho-kinase is an upstream regulator of signaling events leading to phosphorylation of many cytoskeletal proteins in many cell types, we have chosen this kinase as the focus of the present study. In detergent skinned tracheal smooth muscle preparations, we quantified the proteins eluted from the muscle cells over time and monitored the muscle's ability to respond to acetylcholine (ACh) stimulation to produce force and stiffness. In a partially skinned preparation not able to generate active force but could still stiffen upon ACh stimulation, we found that the ACh-induced stiffness was independent of calcium and myosin light chain phosphorylation. This indicates that the myosin light chain-dependent actively cycling crossbridges are not likely the source of the stiffness. The results also indicate that Rho-kinase is central to the ACh-induced stiffness, because inhibition of the kinase by H1152 (1 μM) abolished the stiffening. Furthermore, the rate of relaxation of calcium-induced stiffness in the skinned preparation was faster than that of ACh-induced stiffness, with or without calcium, suggesting that different signaling pathways lead to different means of maintenance of stiffness in the skinned preparation.

DOI10.1152/japplphysiol.00654.2013
Alternate JournalJ. Appl. Physiol.
PubMed ID24072407
PubMed Central IDPMC3841820
Grant ListMOP-13271 / / Canadian Institutes of Health Research / Canada
MOP-37924 / / Canadian Institutes of Health Research / Canada