Regulatory mechanism of smooth muscle contraction studied with gelsolin-treated strips of taenia caeci in guinea pig

Ying Ming Liou, Masaru Watanabe, Masatoshi Yumoto, Shin'ichi Ishiwata

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    The potential roles of the regulatory proteins actin, tropomyosin (Tm), and caldesmon (CaD), i.e., the components of the thin filament, in smooth muscle have been extensively studied in several types of smooth muscles. However, controversy remains on the putative physiological significance of these proteins. In this study, we intended to determine the functional roles of Tm and CaD in the regulation of smooth muscle contraction by using a reconstitution system of the thin filaments. At appropriate conditions, the thin (actin) filaments within skinned smooth muscle strips of taenia caeci in guinea pigs could be selectively removed by an actin-severing protein, gelsolin, without irreversible damage to the contractile apparatus, and then the thin filaments were reconstituted with purified components of thin filaments, i.e., actin, Tm, and CaD. We found that the structural remodeling of actin filaments or thin filaments was functionally linked to the Ca2+-induced force development and reduction in muscle cross-sectional area (CSA). That is, after the reconstitution of the gelsolin-treated skinned smooth muscle strips with pure actin, the Ca2+-dependent force development was partially restored, but the Ca2+-induced reduction in CSA occurred once. In contrast, the reconstitution with actin, followed by Tm and CaD, restored not only the force generation but also both its Ca2+ sensitivity and the reversible Ca2+-dependent reduction in CSA. We confirmed that both removal of the thin filaments by gelsolin treatment and reconstitution of the actin (thin) filaments with Tm and CaD caused no significant changes in the level of myosin regulatory light chain phosphorylation. We thus conclude that Tm and CaD are necessary for the full regulation of smooth muscle contraction in addition to the other regulatory systems, including the myosinlinked one.

    Original languageEnglish
    JournalAmerican Journal of Physiology - Cell Physiology
    Volume296
    Issue number5
    DOIs
    Publication statusPublished - 2009 May

    Fingerprint

    Gelsolin
    Calmodulin-Binding Proteins
    Taenia
    Tropomyosin
    Muscle Contraction
    Smooth Muscle
    Guinea Pigs
    Myosin Light Chains
    Proteins
    Phosphorylation
    Muscles

    Keywords

    • Confocal fluorescence microscopy
    • Force development
    • Phalloidin
    • Phosphorylation of myosin regulatory light chain
    • Reduction in muscle cross-sectional area
    • Thin filament-linked regulation

    ASJC Scopus subject areas

    • Cell Biology
    • Physiology

    Cite this

    Regulatory mechanism of smooth muscle contraction studied with gelsolin-treated strips of taenia caeci in guinea pig. / Liou, Ying Ming; Watanabe, Masaru; Yumoto, Masatoshi; Ishiwata, Shin'ichi.

    In: American Journal of Physiology - Cell Physiology, Vol. 296, No. 5, 05.2009.

    Research output: Contribution to journalArticle

    Liou, Ying Ming ; Watanabe, Masaru ; Yumoto, Masatoshi ; Ishiwata, Shin'ichi. / Regulatory mechanism of smooth muscle contraction studied with gelsolin-treated strips of taenia caeci in guinea pig. In: American Journal of Physiology - Cell Physiology. 2009 ; Vol. 296, No. 5.
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    abstract = "The potential roles of the regulatory proteins actin, tropomyosin (Tm), and caldesmon (CaD), i.e., the components of the thin filament, in smooth muscle have been extensively studied in several types of smooth muscles. However, controversy remains on the putative physiological significance of these proteins. In this study, we intended to determine the functional roles of Tm and CaD in the regulation of smooth muscle contraction by using a reconstitution system of the thin filaments. At appropriate conditions, the thin (actin) filaments within skinned smooth muscle strips of taenia caeci in guinea pigs could be selectively removed by an actin-severing protein, gelsolin, without irreversible damage to the contractile apparatus, and then the thin filaments were reconstituted with purified components of thin filaments, i.e., actin, Tm, and CaD. We found that the structural remodeling of actin filaments or thin filaments was functionally linked to the Ca2+-induced force development and reduction in muscle cross-sectional area (CSA). That is, after the reconstitution of the gelsolin-treated skinned smooth muscle strips with pure actin, the Ca2+-dependent force development was partially restored, but the Ca2+-induced reduction in CSA occurred once. In contrast, the reconstitution with actin, followed by Tm and CaD, restored not only the force generation but also both its Ca2+ sensitivity and the reversible Ca2+-dependent reduction in CSA. We confirmed that both removal of the thin filaments by gelsolin treatment and reconstitution of the actin (thin) filaments with Tm and CaD caused no significant changes in the level of myosin regulatory light chain phosphorylation. We thus conclude that Tm and CaD are necessary for the full regulation of smooth muscle contraction in addition to the other regulatory systems, including the myosinlinked one.",
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