Airway smooth muscle (ASM) has developed a mechanical adaption mechanism by which it transduces force and responds to environmental forces, which is essential for periodic breathing. Cytoskeletal reorganization has been implicated in this process, but the regulatory mechanism remains to be determined. We here observe that ASM abundantly expresses cytoskeleton regulators Limk1 and Limk2, and their expression levels are further upregulated in chronic obstructive pulmonary disease (COPD) animals. By establishing mouse lines with deletions of Limk1 or Limk2, we analyse the length-sensitive contraction, F/G-actin dynamics, and F-actin pool of mutant ASM cells. As LIMK1 phosphorylation does not respond to the contractile stimulation, LIMK1-deficient ASM develops normal maximal force, while LIMK2 or LIMK1/LIMK2 deficient ASMs show approximately 30% inhibition. LIMK2 deletion causes a significant decrease in cofilin phosphorylation along with a reduced F/G-actin ratio. As LIMK2 functions independently of cross-bridge movement, this observation indicates that LIMK2 is necessary for F-actin dynamics and hence force transduction. Moreover, LIMK2-deficient ASMs display abolishes stretching-induced suppression of 5-hydroxytryptamine (5-HT) but not acetylcholine-evoks force, which is due to the differential contraction mechanisms adopted by the agonists. We propose that LIMK2-mediated cofilin phosphorylation is required for membrane cytoskeleton reorganization that is necessary for ASM mechanical adaption including the 5-HT-evoked length-sensitive effect.
|
Aoun, J., Mayne, K., Baeck, J., Sanders, K., Ward, S., Greenwood, I., Bulley, S., Jaggar, J., Earley, S., Leblanc, N., 2020. Ano1, cav1.2 and ip3r form a functional unit of excitation-contraction coupling during agonist-mediated contraction of mouse pulmonary arterial smooth muscle. Biophys. J. 118, 563a-564a.
|
|
Chen, C.P., Chen, X., Qiao, Y.N., Wang, P., He, W.Q., Zhang, C.H., Zhao, W., Gao, Y.Q., Chen, C., Tao, T., et al., 2015. In vivo roles for myosin phosphatase targeting subunit-1 phosphorylation sites T694 and T852 in bladder smooth muscle contraction. J. Physiol. 593, 681-700.
|
|
Draeger, A., Amos, W.B., Ikebe, M., Small, J.V., 1990. The cytoskeletal and contractile apparatus of smooth muscle: contraction bands and segmentation of the contractile elements. J. Cell Biol. 111, 2463-2473.
|
|
Gabella, G., 1984. Structural apparatus for force transmission in smooth muscles. Physiol. Rev. 64, 455-477.
|
|
Gunst, S.J., Meiss, R.A., Wu, M.F., Rowe, M., 1995. Mechanisms for the mechanical plasticity of tracheal smooth muscle. Am. J. Physiol. Cell Physiol. 268, C1267-C1276.
|
|
Gunst, S.J., Shen, X., Ramchandani, R., Tepper, R.S., 2001. Bronchoprotective and bronchodilatory effects of deep inspiration in rabbits subjected to bronchial challenge. J. Appl. Physiol. 91, 2511-2516.
|
|
Gunst, S.J., Stropp, J.Q., Service, J., 1990. Mechanical modulation of pressurevolume characteristics of contracted canine airways in vitro. J. Appl. Physiol. 68, 2223-2229.
|
|
Gunst, S.J., Tang, D.D., 2000. The contractile apparatus and mechanical properties of airway smooth muscle. Eur. Respir. J. 15, 600.
|
|
Gunst, S.J., Tang, D.D., Opazo Saez, A., 2003. Cytoskeletal remodeling of the airway smooth muscle cell: a mechanism for adaptation to mechanical forces in the lung. Respir. Physiol. Neurobiol. 137, 151-168.
|
|
Gunst, S.J., Zhang, W., 2008. Actin cytoskeletal dynamics in smooth muscle: a new paradigm for the regulation of smooth muscle contraction. Am. J. Physiol. Cell Physiol. 295, C576.
|
|
He, W.-Q., Qiao, Y.-N., Zhang, C.-H., Peng, Y.-J., Chen, C., Wang, P., Gao, Y.-Q., Chen, C., Chen, X., Tao, T., et al., 2011. Role of myosin light chain kinase in regulation of basal blood pressure and maintenance of salt-induced hypertension. Am. J. Physiol. Heart Circ. Physiol. 301, H584-H591.
|
|
He, W.Q., Qiao, Y.N., Peng, Y.J., Zha, J.M., Zhang, C.H., Chen, C., Chen, C.P., Wang, P., Yang, X., Li, C.J., et al., 2013. Altered contractile phenotypes of intestinal smooth muscle in mice deficient in myosin phosphatase target subunit 1. Gastroenterology 144, 1456-1465.
|
|
Hirshman, C.A., Togashi, H., Shao, D., Emala, C.W., 1998. Gai-2 is required for carbachol-induced stress fiber formation in human airway smooth muscle cells. Am. J. Physiol. Lung Cell Mol. Physiol. 275, L911-L916.
|
|
Hoover, W.C., Zhang, W., Xue, Z., Gao, H., Chernoff, J., Clapp, D.W., Gunst, S.J., Tepper, R.S., 2012. Inhibition of p21 activated kinase (pak) reduces airway responsiveness in vivo and in vitro in murine and human airways. PLoS One 7, e42601.
|
|
Horowitz, A., Menice, C.B., Laporte, R., Morgan, K.G., 1996. Mechanisms of smooth muscle contraction. Physiol. Rev. 76, 967-1003.
|
|
Huang, Y., Zhang, W., Gunst, S.J., 2010. Activation of vinculin induced by cholinergic stimulation regulates contraction of tracheal smooth muscle tissue. J. Biol. Chem. 286, 3630-3644.
|
|
Jin, X., Shah, S., Du, X., Zhang, H., Gamper, N., 2016. Activation of Ca2+-activated Cl- channel ano1 by localized Ca2+ signals. J. Physiol. 594, 19-30.
|
|
Jin, X., Shah, S., Liu, Y., Zhang, H., Lees, M., Fu, Z., Lippiat, J.D., Beech, D.J., Sivaprasadarao, A., Baldwin, S.A., et al., 2013. Activation of the Cl- channel ANO1 by localized calcium signals in nociceptive sensory neurons requires coupling with the IP3 receptor. Sci. Signal. 6, ra73.
|
|
Kamm, K.E., Stull, J.T., 1985. The function of myosin and myosin light chain kinase phosphorylation in smooth muscle. Annu. Rev. Pharmacol. Toxicol. 25, 593-620.
|
|
Lee, S.S., von Der Weid, P.-Y., 2013. Role of voltage-dependent calcium channels in stretch-induced lymphatic vessel contractions. Faseb. J. 27, 681.611.
|
|
Manetti, F., 2012. Lim kinases are attractive targets with many macromolecular partners and only a few small molecule regulators. Med. Res. Rev. 32, 968-998.
|
|
Mehta, D., Gunst, S.J., 1999. Actin polymerization stimulated by contractile activation regulates force development in canine tracheal smooth muscle. J. Physiol. 519(Pt 3), 829-840.
|
|
Mizuno, K., 2013. Signaling mechanisms and functional roles of cofilin phosphorylation and dephosphorylation. Cell Signal. 25, 457-469.
|
|
Nadel, J.A., Tierney, D.F., 1961. Effect of a previous deep inspiration on airway resistance in man. J. Appl. Physiol. 16, 717-719.
|
|
Obara, K., Yabu, H., 1994. Effect of cytochalasin b on intestinal smooth muscle cells. Eur. J. Pharmacol. 255, 139-147.
|
|
Ohashi, K., 2015. Roles of cofilin in development and its mechanisms of regulation. Dev. Growth Differ. 57, 275-290.
|
|
Postma, D.S., Kerstjens, H.A.M., 1998. Characteristics of airway hyperresponsiveness in asthma and chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 158, S187-S192.
|
|
Prakash, Y.S., 2016. Emerging concepts in smooth muscle contributions to airway structure and function: implications for health and disease. Am. J. Physiol. Lung Cell Mol. Physiol. 311, L1113-L1140.
|
|
Prunier, C., Prudent, R., Kapur, R., Sadoul, K., Lafanechère, L., 2017. Lim kinases: cofilin and beyond. Oncotarget 8, 41749-41763.
|
|
Ran, F.A., Hsu, P.D., Lin, C.-Y., Gootenberg, J.S., Konermann, S., Trevino, A.E., Scott, D.A., Inoue, A., Matoba, S., Zhang, Y., et al., 2013. Double nicking by RNAguided CRISPR Cas9 for enhanced genome editing specificity. Cell 154, 1380-1389.
|
|
Riedl, J., Crevenna, A.H., Kessenbrock, K., Yu, J.H., Neukirchen, D., Bista, M., Bradke, F., Jenne, D., Holak, T.A., Werb, Z., et al., 2008. Lifeact: a versatile marker to visualize f-actin. Nat. Methods 5, 605-607.
|
|
Saito, S.Y., Hori, M., Ozaki, H., Karaki, H., 1996. Cytochalasin D inhibits smooth muscle contraction by directly inhibiting contractile apparatus. J. Smooth Muscle Res. 32, 51-60.
|
|
Schaller, M.D., 2010. Cellular functions of FAK kinases: insight into molecular mechanisms and novel functions. J. Cell Sci. 123, 1007-1013.
|
|
Scott, R.W., Olson, M.F., 2007. LIM kinases: function, regulation and association with human disease. J. Mol. Med. 85, 555-568.
|
|
Shen, X., Gunst, S.J., Tepper, R.S., 1997. Effect of tidal volume and frequency on airway responsiveness in mechanically ventilated rabbits. J. Appl. Physiol. (1985) 83, 1202-1208.
|
|
Siegman, M.J., Butler, T.M., Mooers, S.U., Davies, R.E., 1976. Calcium-dependent resistance to stretch and stress relaxation in resting smooth muscles. Am. J. Physiol. 231, 1501-1508.
|
|
Skloot, G., Togias, A., 2003. Bronchodilation and bronchoprotection by deep inspiration and their relationship to bronchial hyperresponsiveness. Clin. Rev. Allergy Immunol. 24, 55-71.
|
|
Small, J.V., 1995. Structure-function relationships in smooth muscle: the missing links. Bioessays 17, 785-792.
|
|
Tang, D., Mehta, D., Gunst, S.J., 1999. Mechanosensitive tyrosine phosphorylation of paxillin and focal adhesion kinase in tracheal smooth muscle. Am. J. Physiol. 276, C250-C258.
|
|
Tang, D.D., Gunst, S.J., 2001. Depletion of focal adhesion kinase by antisense depresses contractile activation of smooth muscle. Am. J. Physiol. Cell Physiol. 280, C874-C883.
|
|
Thirstrup, S., 2000. Control of airway smooth muscle tone. I - electrophysiology and contractile mediators. Respir. Med. 94, 328-336.
|
|
Wang, P., Zhao, W., Sun, J., Tao, T., Chen, X., Zheng, Y.Y., Zhang, C.H., Chen, Z., Gao, Y.Q., She, F., et al., 2018. Inflammatory mediators mediate airway smooth muscle contraction through a G protein-coupled receptor-transmembrane protein 16A-voltage-dependent Ca2+ channel axis and contribute to bronchial hyperresponsiveness in asthma. J. Allergy Clin. Immunol. 141, 1259-1268 e1211.
|
|
Wong, R.S., Larcombe, A.N., Fernandes, L.B., Zosky, G.R., Noble, P.B., 2012. The mechanism of deep inspiration-induced bronchoprotection: evidence from a mouse model. Eur. Respir. J. 40, 982-989.
|
|
Xu, J., Zhao, M., Liao, S., 2000. Establishment and pathological study of models of chronic obstructive pulmonary disease by SO2 inhalation method. Chin. Med. J.(Engl) 113, 213-216.
|
|
Zhang, W.-C., Peng, Y.-J., Zhang, G.-S., He, W.-Q., Qiao, Y.-N., Dong, Y.-Y., Gao, Y.-Q., Chen, C., Zhang, C.-H., Li, W., et al., 2010. Myosin light chain kinase is necessary for tonic airway smooth muscle contraction. J. Biol. Chem. 285, 5522-5531.
|
|
Zhang, W., Gunst, S.J., 2008. Interactions of airway smooth muscle cells with their tissue matrix: implications for contraction. Proc. Am. Thorac. Soc. 5, 32-39.
|
|
Zhang, W., Gunst, S.J., 2019. Molecular mechanisms for the mechanical modulation of airway responsiveness. J. Eng. Sci. Med. Diagnost. Ther. 2, 0108051-0108058.
|
|
Zhao, R., Du, L., Huang, Y., Wu, Y., Gunst, S.J., 2008. Actin depolymerization factor/cofilin activation regulates actin polymerization and tension development in canine tracheal smooth muscle. J. Biol. Chem. 283, 36522-36531.
|
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Supported by: Beijing Renhe Information Technology Co. Ltd
Yeqiong Li, Yuwei Zhou, Pei Wang, Tao Tao, Lisha Wei, Ye Wang, Wei Wang, Yanyan Zheng, Zhihui Jiang, Tiantian Qiu, Wei Zhao, Jie Sun, Xin Chen, Xue-Na Zhang, Min-Sheng Zhu. LIMK2 is required for membrane cytoskeleton reorganization of contracting airway smooth muscle[J]. Journal of Genetics and Genomics, 2021, 48(6): 452-462. doi: 10.1016/j.jgg.2021.04.014
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