F-actin stabilization increases tension cost during contraction of permeabilized airway smooth muscle in dogs.
1. Dynamic actin reorganization involving actin polymerization and depolymerization may play an important functional role in smooth muscle. 2. This study tested the hypothesis that F-actin stabilization by phalloidin increases tension cost (i.e. ATP hydrolysis rate per unit of isometric force) during Ca2+-induced activation of Triton X-100-permeabilized canine tracheal smooth muscle. 3. Adenosine 5'-triphosphate (ATP) hydrolysis rate was quantified using an enzyme-coupled NADH fluorometric technique, regulatory myosin light chain (rMLC) phosphorylation was measured by Western blot analysis, and maximum unloaded shortening velocity (Vmax) was estimated by interpolation of the force-velocity relationship to zero load during isotonic loading. 4. Maximal activation with 10 microM free Ca2+ induced sustained increases in isometric force, stiffness, and rMLC phosphorylation. However, the increase in ATP hydrolysis rate initially reached peak values, but then declined to steady-state levels above that of the unstimulated muscle. Thus, tension cost decreased throughout steady-state isometric force. 5. Following incubation of permeabilized strips with 50 microM phalloidin for 1 h, the increases in isometric force and stiffness were not sustained despite a sustained increase in rMLC phosphorylation. Also, after an initial decline, tension cost increased throughout activation. Phalloidin had no effect on Vmax during steady-state isometric force or on rMLC phosphorylation. 6. These findings suggest that dynamic reorganization of actin is necessary for optimal energy utilization during contraction of permeabilized airway smooth muscle.
Jones
K A
KA
Departments of Anesthesiology, and Physiology and Biophysics, Mayo Clinic and Mayo Foundation, Rochester, MN 55905, USA. kjones@mayo.edu
Perkins
W J
WJ
Lorenz
R R
RR
Prakash
Y S
YS
Sieck
G C
GC
Warner
D O
DO
D000199:Actins / Q000378:metabolism / Q000502:physiology*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000502:physiology; D004285:Dogs; D004558:Electric Stimulation; D004734:Energy Metabolism / Q000187:drug effects / Q000502:physiology; D005260:Female; D006868:Hydrolysis; D066298:In Vitro Techniques; D007537:Isometric Contraction / Q000187:drug effects / Q000502:physiology; D008297:Male; D008856:Microscopy, Fluorescence; D009119:Muscle Contraction / Q000502:physiology; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D018994:Myosin Light Chains / Q000378:metabolism; D017830:Octoxynol / Q000494:pharmacology; D010590:Phalloidine / Q000494:pharmacology; D010766:Phosphorylation; D014132:Trachea / Q000187:drug effects / Q000502:physiology
D000199:Actins; D018994:Myosin Light Chains; D010590:Phalloidine; D000255:Adenosine Triphosphate; D017830:Octoxynol; D002118:Calcium