title: ROCK1 translocates from non-caveolar to caveolar regions upon KCl stimulation in airway smooth muscle.
abstract: Airway smooth muscle (ASM) membrane depolarization through KCl opens L-type voltage dependent Ca2+ channels (Ca(v)1.2); its opening was considered the cause of KCl contraction. This substance is used to bypass intracellular second messenger pathways. It is now clear that KCl also activates RhoA/Rho kinase (ROCK) pathway. ROCK isoforms are characterized as ROCK1 and ROCK2. Because ROCK1 seems the most abundant isotype in lung, we studied its participation in KCl stimulated bovine ASM. With methyl-beta-cyclodextrin (MbetaCD) we disrupted caveolae, a membrane compartment considered as the RhoA/ROCK assembly site, and found that KCl contraction was reduced to the same extent (~26%) as Y-27632 (ROCK inhibitor) treated tissues. We confirmed that KCl induces ROCK activation and this effect was annulled by Y-27632 or MbetaCD. In isolated plasmalemma, ROCK1 was localized in non-caveolar membrane fractions in Western blots from control tissues, but it transferred to caveolae in samples from tissues stimulated with KCl. Ca(v)1.2 was found at the non-caveolar membrane fractions in control and MbetaCD treated tissues. In MbetaCD treated tissues stimulated with KCl, contraction was abolished by nifedipine; only the response to Ca(v)1.2 opening remained as the ROCK component disappeared. Our results show that, in ASM, the KCl contraction involves the translocation of ROCK1 from non-caveolar to caveolar regions and that the proper physiological response depends on this translocation.
authors.affiliation: Department of Bronchial Hyperreactivity Research, National Institute of Respiratory Diseases, Mexico City, Mexico. bsommerc@hotmail.com.
authors.forename: B
authors.initials: B
authors.lastname: Sommer
authors.forename: L M
authors.initials: LM
authors.lastname: Montano
authors.forename: J
authors.initials: J
authors.lastname: Chávez
authors.forename: V
authors.initials: V
authors.lastname: Carbajal
authors.forename: L M
authors.initials: LM
authors.lastname: García-Hernandez
authors.forename: C
authors.initials: C
authors.lastname: Irles
authors.forename: A M
authors.initials: AM
authors.lastname: Jiménez-Garduno
authors.forename: A
authors.initials: A
authors.lastname: Ortega
mesh_terms: D000818:Animals; D002417:Cattle; D021941:Caveolae / Q000187:drug effects / Q000378:metabolism*; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009125:Muscle Relaxants, Central / Q000494:pharmacology; D009130:Muscle, Smooth / Q000187:drug effects / Q000378:metabolism*; D009924:Organ Culture Techniques; D011189:Potassium Chloride / Q000494:pharmacology*; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D014132:Trachea / Q000187:drug effects / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: D009125:Muscle Relaxants, Central; D011189:Potassium Chloride; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases