title: Shear stress-induced Ang II AT1 receptor activation: G-protein dependent and independent mechanisms.
abstract: Mechanotransduction enables cells to sense and respond to stimuli, such as strain, pressure and shear stress (SS), critical for maintenance of cardiovascular homeostasis or pathological states. The angiotensin II type 1 receptor (AT1R) was the first G protein-coupled receptor described to display stretch-induced activation in cardiomyocytes independent of its ligand Ang II. Here, we assessed whether SS (15 dynes/cm(2), 10 min), an important mechanical force present in the cardiovascular system, activates AT1R independent of its ligand. SS induced extracellular signal-regulated kinase (ERK) activation, used as a surrogate of AT1R activation, in Chinese hamster ovary cells expressing the AT1R (CHO+AT1) but not in wild type cells (CHO). AT1R dependent SS-induced ERK activation involves Ca(2+) inflow and activation of Gαq since Ca(2+) chelator EGTA or Gαq-specific inhibitor YM-254890 decreased SS-induced ERK activation. On the other hand, the activation of JAK-2 and Src, two intracellular signaling molecules independent of G protein activation, were not differently modulated in the presence of AT1R. Also, ERK activation by SS was observed in CHO cells expressing the mutated AT1R DRY/AAY, which has impaired ability to activate Gαq dependent intracellular signaling. Altogether we provided evidence that SS activates AT1R in the absence of its ligand by both a G protein-dependent and -independent pathways. The biological relevance of these observations deserves to be further investigated since the novel mechanisms described extend the knowledge of the activation of GPCRs independent of its traditional ligand.
authors.affiliation: Laboratory of Genetics and Molecular Cardiology, Heart Institute, InCor, University of São Paulo Medical School, São Paulo 05403-000, Brazil.
authors.forename: Valerio G
authors.initials: VG
authors.lastname: Barauna
authors.forename: Flávio C
authors.initials: FC
authors.lastname: Magalhaes
authors.forename: Luciene C G
authors.initials: LC
authors.lastname: Campos
authors.forename: Rosana I
authors.initials: RI
authors.lastname: Reis
authors.forename: Satya P
authors.initials: SP
authors.lastname: Kunapuli
authors.forename: Claudio M
authors.initials: CM
authors.lastname: Costa-Neto
authors.forename: Ayumi A
authors.initials: AA
authors.lastname: Miyakawa
authors.forename: Jose E
authors.initials: JE
authors.lastname: Krieger
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000502:physiology*; D016133:Polymerase Chain Reaction; D044140:Receptor, Angiotensin, Type 1 / Q000378:metabolism*; D013312:Stress, Physiological
chemical_list: C544065:AGTR1 protein, human; D044140:Receptor, Angiotensin, Type 1; D048049:Extracellular Signal-Regulated MAP Kinases; D019204:GTP-Binding Proteins