Rho-kinase and contractile apparatus proteins in murine airway hyperresponsiveness

Mapping physiological G protein-coupled receptor signaling pathways reveals a role for receptor phosphorylation in airway contraction

“…In these experiments, WT mice sensitized with ovalbumin and challenged with saline (0.9% NaCl) showed an increase in lung resistance to ACh that was significantly augmented in mice sensitized with ovalbumin and challenged with ovalbumin (Fig. 4C), a response defined as classical airway hyperresponsiveness (24,26,27). In contrast, M3-KI mice sensitized with ovalbumin and challenged with ovalbumin showed no hyperresponsiveness in response to ACh administration (Fig.…”

“…M3-mAChR signaling through RhoA in ASM has been closely linked with experimentally induced airway hyperresponsiveness in different studies using murine models of allergy (23,24). We tested here if the reduced coupling of the phosphodeficient M3-mAChR mutant to RhoA signaling and the subsequent reduction in ASM contraction observed in M3-KI mice had an impact on allergeninduced airway hyperresponsiveness using a murine model of allergic inflammatory airway disease (25).…”

The importance of complete tissue homogenization for accurate stoichiometric measurement of myosin light chain phosphorylation in airway smooth muscle

“…A review of the recent literature revealed a wide range of techniques that are being used for MLC20 phosphorylation measurements in airway smooth muscle. These include: isoelectric focusing followed by Coomassie brilliant blue staining (An and Hai 2000;Pabelick et al 2000) or colloidal gold labeling (Parris et al 1999); sodium dodecyl sulfate (SDS) acrylamide gel electrophoresis followed by Western blot with an antibody specific to phosphorylation site Ser19 (Gil et al 2006;Khromov et al 2006;Smith et al 2007;Fairbank et al 2008;Goto et al 2008;Morin et al 2008;Witzenrath et al 2008;Lee et al 2009;Wang et al 2014); Phos-tag gels initially developed for small blood vessels (Takeya et al 2008) but recently used for airway smooth muscle (Matusovsky et al 2014), and the classic urea-glycerol gel, which we chose as the method to obtain the stoichiometry of MLC20 phosphorylation in tracheal smooth muscle strips.…”

Tratamento com inibidor da Rho quinase em cobaias com inflamação alérgica crônica: modulação da inflamação eosinofílica, da expressão de citocinas inflamatórias, da matriz extracelular, do estresse oxidativo e da reatividade de vias aéreas

“…De fato, a relação entre Rho quinase e hiperresponsividade das vias aéreas já foi demonstrada em diferentes estudos (Chiba et al, 1999;Schaafsma et al, 2004;Witzenrath et al, 2008). Inibindo a sensibilização do Ca 2+ , o Y-27632 é capaz de relaxar completamente o músculo liso das vias aéreas (Yoshii et al, 1999;Iizuka et al, 2000).…”

“…Os efeitos da inibição aguda da Rho quinase em animais sensibilizados foram analisados em alguns estudos (Schaafsma et al, 2008;Fernandes et al, 2006;Zhang et al, 1995;Henry et al, 2005;Witzenrath et al, 2008;Hashimoto et al, 2001;Hanazaki et al, 2000;Hanazaki et al, 2008). Schaafsma et al (2008) Além disso, é mostrado claramente que a ativação da iNOS pode contribuir para a formação de peroxinitrito, que é um agente oxidante formado pela interação entre NO e superóxido.…”