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J Appl Physiol 98: 489-497, 2005. First published October 1, 2004; doi:10.1152/japplphysiol.01378.2003
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Latrunculin B increases force fluctuation-induced relengthening of ACh-contracted, isotonically shortened canine tracheal smooth muscle

M. L. Dowell,1 O. J. Lakser,2 W. T. Gerthoffer,3 J. J. Fredberg,4 G. L. Stelmack,5 A. J. Halayko,5 J. Solway,1,* and R. W. Mitchell1,*

1Section of Pulmonary and Critical Care Medicine, University of Chicago, and 2Children's Memorial Hospital and Northwestern University, Chicago, Illinois; 3Department of Pharmacology, University of Nevada School of Medicine, Reno, Nevada; 4Physiology Program, Harvard School of Public Health, Boston, Massachusetts; and 5Section of Respiratory Diseases, University of Manitoba, Winnipeg, Manitoba, Canada

Submitted 22 December 2003 ; accepted in final form 28 September 2004

We hypothesized that differences in actin filament length could influence force fluctuation-induced relengthening (FFIR) of contracted airway smooth muscle and tested this hypothesis as follows. One-hundred micromolar ACh-stimulated canine tracheal smooth muscle (TSM) strips set at optimal reference length (Lref) were allowed to shorten against 32% maximal isometric force (Fmax) steady preload, after which force oscillations of ±16% Fmax were superimposed. Strips relengthened during force oscillations. We measured hysteresivity and calculated FFIR as the difference between muscle length before and after 20-min imposed force oscillations. Strips were relaxed by ACh removal and treated for 1 h with 30 nM latrunculin B (sequesters G-actin and promotes depolymerization) or 500 nM jasplakinolide (stabilizes actin filaments and opposes depolymerization). A second isotonic contraction protocol was then performed; FFIR and hysteresivity were again measured. Latrunculin B increased FFIR by 92.2 ± 27.6% Lref and hysteresivity by 31.8 ± 13.5% vs. pretreatment values. In contrast, jasplakinolide had little influence on relengthening by itself; neither FFIR nor hysteresivity was significantly affected. However, when jasplakinolide-treated tissues were then incubated with latrunculin B in the continued presence of jasplakinolide for 1 more h and a third contraction protocol performed, latrunculin B no longer substantially enhanced TSM relengthening. In TSM treated with latrunculin B + jasplakinolide, FFIR increased by only 3.03 ± 5.2% Lref and hysteresivity by 4.14 ± 4.9% compared with its first (pre-jasplakinolide or latrunculin B) value. These results suggest that actin filament length, in part, determines the relengthening of contracted airway smooth muscle.

actin filament dynamics; force oscillations; isotonic contractions; hysteresivity


Address for reprint requests and other correspondence: R. W. Mitchell, Univ. of Chicago, MC6026, 5841 S. Maryland Ave., Chicago, IL 60637 (E-mail: rmitchel{at}medicine.bsd.uchicago.edu)




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