title: Repeated allergen inhalation induces cytoskeletal remodeling in smooth muscle from rat bronchioles.
abstract: Airway hyperresponsiveness (AHR) is associated with airway wall structural remodeling and alterations in airway smooth muscle (ASM) function. Previously, in bronchioles from Brown Norway rats challenged by repeated ovalbumin (OVA) inhalation, we have reported increased force generation and depletion of smooth muscle contractile proteins. Here, we investigated if cytoskeletal changes in smooth muscle could account for this paradox. Sensitized rats (n = 5/group) were repeatedly challenged with OVA or saline, and the lungs were removed 24 h after the last challenge. Levels of globular (G) and filamentous (F) actin in bronchioles were determined by DNase I inhibition and contraction assessed in intact small bronchioles using a myograph. DNase I inhibition assays showed that G-actin monomers were more abundant ( approximately 1F:2G) in extracts from resting small bronchioles from OVA- or saline-challenged animals. However, while contractile protein levels in bronchioles were reduced by OVA (P < 0.05), the proportion of F:G actin was 1.8-fold greater compared with saline challenge (P < 0.05). Consistent with induction of F-actin after OVA challenge, increases in maximum tension development to carbachol or KCl in small bronchioles from OVA-challenged animals were abrogated (P < 0.01) by actin cytoskeleton disruption with 0.5 microM latrunculin A. Cytoskeletal stabilization of F-actin with 0.1 microM jasplakinolide potentiated maximum contractions to carbachol or KCl (P < 0.05) in bronchioles from OVA- but not saline-treated rats. We conclude that alterations in the composition and/or arrangement of the contractile apparatus after OVA exposure confer enhanced contractile responses, possibly as a result of increased F-actin content. Such a mechanism may have relevance for AHR found in allergic asthma.
authors.affiliation: King's College London School of Medicine, MRC & Asthma UK Centre in Allergic Mechanisms of Asthma, Division of Asthma, Allergy and Lung Biology, London, United kingdom.
authors.forename: Clare G
authors.initials: CG
authors.lastname: McVicker
authors.forename: Sum-Yee
authors.initials: SY
authors.lastname: Leung
authors.forename: Varsha
authors.initials: V
authors.lastname: Kanabar
authors.forename: Lyn M
authors.initials: LM
authors.lastname: Moir
authors.forename: Katharina
authors.initials: K
authors.lastname: Mahn
authors.forename: K Fan
authors.initials: KF
authors.lastname: Chung
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Hirst
mesh_terms: D000199:Actins / Q000378:metabolism; D000485:Allergens / Q000276:immunology*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001249:Asthma / Q000276:immunology; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D001980:Bronchi / Q000166:cytology* / Q000187:drug effects / Q000276:immunology*; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D047630:Depsipeptides / Q000494:pharmacology; D019570:Inhalation Exposure; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010047:Ovalbumin / Q000008:administration & dosage / Q000276:immunology; D051381:Rats; D053778:Thiazolidines / Q000494:pharmacology
chemical_list: D000199:Actins; D000485:Allergens; D000970:Antineoplastic Agents; D019086:Bridged Bicyclo Compounds, Heterocyclic; D047630:Depsipeptides; D053778:Thiazolidines; C057531:jasplakinolide; D002217:Carbachol; D010047:Ovalbumin; C037067:latrunculin A