Force maintenance and myosin filament assembly regulated by Rho-kinase in airway smooth muscle

Lan, Bo; Deng, Linhong; Donovan, Graham M.; Chin, Leslie Y. M.; Syyong, Harley T.; Wang, Lu; Zhang, Jenny; Pascoe, Chris D.; Norris, Brandon A.; Liu, Jeffrey C.-Y.; Swyngedouw, Nicholas E.; Banaem, Saleha M.; Paré, Peter D.; Seow, Chun Y.

doi:10.1152/ajplung.00222.2014

Cited by 38 publications

(41 citation statements)

References 25 publications

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“…In slower nonmuscle contractile processes, order may be capable of emerging spontaneously through self-organization. We note, however, that ROCK localization has not been determined in smooth muscle where ROCK is known to play a role in calcium sensitization and force maintenance (Lan et al, 2015; Uehata et al, 1997). Our data suggests that ROCK localization could support its function in other contexts.…”

Section: Discussionmentioning

confidence: 82%

Apical Sarcomere-like Actomyosin Contracts Nonmuscle Drosophila Epithelial Cells

Coravos

Martin

2016

Developmental Cell

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Summary Actomyosin networks generate contractile force that changes cell and tissue shape. In muscle cells, actin filaments and myosin II appear in a polarized structure called a sarcomere, where myosin II is localized in the center. Nonmuscle cortical actomyosin networks are thought to contract when nonmuscle myosin II (myosin) is activated throughout a mixed-polarity actin network. Here, we identified a mutant version of the myosin-activating kinase, ROCK, that localizes diffusely, rather than centrally, in epithelial cell apices. Surprisingly, this mutant inhibits constriction, suggesting that centrally localized apical ROCK/myosin activity promotes contraction. We determined actin cytoskeletal polarity by developing a barbed end incorporation assay for Drosophila embryos, which revealed barbed end enrichment at junctions. Our results demonstrate that epithelial cells contract with a spatially organized apical actomyosin cortex, involving a polarized actin cytoskeleton and centrally positioned myosin, with cell-scale order that resembles a muscle sarcomere.

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Section: Discussionmentioning

confidence: 82%

Apical Sarcomere-like Actomyosin Contracts Nonmuscle Drosophila Epithelial Cells

Coravos

Martin

2016

Developmental Cell

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“…The data suggest that ROCK2-mediated events are downstream of O 3 -induced inflammation and/or injury. Although ROCK is known to contribute to ASM contraction (4,7,13,18,35,37,71), the specific role of the ROCK2 isoform has not been established. Consequently, we examined the effect of a ROCK2 inhibitor on contractile force of HASM cells using FTTM.…”

Section: Rock2 Inhibition Decreases Contractile Force Of Hasm Cellsmentioning

confidence: 99%

ROCK insufficiency attenuates ozone-induced airway hyperresponsiveness in mice

Kasahara¹,

Mathews²,

Park³

et al. 2015

American Journal of Physiology-Lung Cellular and Molecular Phys

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Ozone causes airway hyperresponsiveness (AHR) and pulmonary inflammation. Rho kinase (ROCK) is a key regulator of smooth muscle cell contraction and inflammatory cell migration. To determine the contribution of the two ROCK isoforms ROCK1 and ROCK2 to ozone-induced AHR, we exposed wild-type, ROCK1(+/-), and ROCK2(+/-) mice to air or ozone (2 ppm for 3 h) and evaluated mice 24 h later. ROCK1 or ROCK2 haploinsufficiency did not affect airway responsiveness in air-exposed mice but significantly reduced ozone-induced AHR, with a greater reduction in ROCK2(+/-) mice despite increased bronchoalveolar lavage (BAL) inflammatory cells in ROCK2(+/-) mice. Compared with wild-type mice, ozone-induced increases in BAL hyaluronan, a matrix protein implicated in ozone-induced AHR, were lower in ROCK1(+/-) but not ROCK2(+/-) mice. Ozone-induced increases in other inflammatory moieties reported to contribute to ozone-induced AHR (IL-17A, osteopontin, TNFα) were not different in wild-type vs. ROCK1(+/-) or ROCK2(+/-) mice. We also observed a dose-dependent reduction in ozone-induced AHR after treatment with the ROCK1/ROCK2 inhibitor fasudil, even though fasudil was administered after induction of inflammation. Ozone increased pulmonary expression of ROCK2 but not ROCK1 or RhoA. A ROCK2 inhibitor, SR3677, reduced contractile forces in primary human airway smooth muscle cells, confirming a role for ROCK2 in airway smooth muscle contraction. Our results demonstrate that ozone-induced AHR requires ROCK. Whereas ROCK1-dependent changes in hyaluronan may contribute to ROCK1's role in O3-induced AHR, the role of ROCK2 is downstream of inflammation, likely at the level of airway smooth muscle contraction.

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“…Another limitation of the study is that we only focused on the pathways related to the regulation of contraction through actomyosin interaction. During the prolonged (120 min) contraction induced by Bay K8644, other pathways could have come into play, notably the pathways related to the regulation of actin cytoskeletal function and the muscle’s ability to maintain force through non-cross-bridge mechanisms (Lehman and Morgan, 2012; Yamin and Morgan, 2012; Lan et al, 2015). Both intracellular calcium and MLC phosphorylation levels are known to vary with time during contraction (Dillon et al, 1981).…”

Section: Discussionmentioning

confidence: 99%

Sustained Contraction in Vascular Smooth Muscle by Activation of L-type Ca2+ Channels Does Not Involve Ca2+ Sensitization or Caldesmon

2016

Self Cite

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Vascular smooth muscle (VSM) is unique in its ability to maintain an intrinsic level of contractile force, known as tone. Vascular tone is believed to arise from the constitutive activity of membrane-bound L-type Ca2+ channels (LTCC). This study used a pharmacological agonist of LTCC, Bay K8644, to elicit a sustained, sub-maximal contraction in VSM that mimics tone. Downstream signaling was investigated in order to determine what molecules are responsible for tone. Medial strips of swine carotid artery were stimulated with 100 nM Bay K8644 to induce a sustained level of force. Force and phosphorylation levels of myosin light chain (MLC), MAP kinase, MYPT1, CPI-17, and caldesmon were measured during Bay K8644 stimulation in the presence and absence of nifedipine, ML-7, U0126, bisindolylmaleimide (Bis), and H-1152. Nifedipine and ML-7 inhibited force and MLC phosphorylation in response to Bay K8644. Inhibition of Rho kinase (H-1152) but not PKC (Bis) inhibited Bay K8644 induced force. U0126 significantly increased Bay K8644-dependent force with no effect on MLC phosphorylation. Neither CPI-17 nor caldesmon phosphorylation were increased during the maintenance of sustained force. Our results suggest that force due to the influx of calcium through LTCCs is partially MLC phosphorylation-dependent but does not involve PKC or caldesmon. Interestingly, inhibition of MLC kinase (MLCK) and PKC significantly increased MAP kinase phosphorylation suggesting that MLCK and PKC may directly or indirectly inhibit MAP kinase activity during prolonged contractions induced by Bay K8544.

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Force maintenance and myosin filament assembly regulated by Rho-kinase in airway smooth muscle

Cited by 38 publications

References 25 publications

Apical Sarcomere-like Actomyosin Contracts Nonmuscle Drosophila Epithelial Cells

Apical Sarcomere-like Actomyosin Contracts Nonmuscle Drosophila Epithelial Cells

ROCK insufficiency attenuates ozone-induced airway hyperresponsiveness in mice

Sustained Contraction in Vascular Smooth Muscle by Activation of L-type Ca2+ Channels Does Not Involve Ca2+ Sensitization or Caldesmon

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