title: An alpha1A-adrenergic-extracellular signal-regulated kinase survival signaling pathway in cardiac myocytes.
abstract: BACKGROUND
  In alpha1-AR knockout (alpha1ABKO) mice that lacked cardiac myocyte alpha1-adrenergic receptor (alpha1-AR) binding, aortic constriction induced apoptosis, dilated cardiomyopathy, and death. However, it was unclear whether these effects were attributable to a lack of cardiac myocyte alpha1-ARs and whether the alpha1A, alpha1B, or both subtypes mediated protection. Therefore, we investigated alpha1A and alpha1B subtype-specific survival signaling in cultured cardiac myocytes to test for a direct protective effect of alpha1-ARs in cardiac myocytes.
  
  
  RESULTS
  We cultured alpha1ABKO myocytes and reconstituted alpha1-AR signaling with adenoviruses expressing alpha1-GFP fusion proteins. Myocyte death was induced by norepinephrine, doxorubicin, or H2O2 and was measured by annexin V/propidium iodide staining. In alpha1ABKO myocytes, all 3 stimuli significantly increased apoptosis and necrosis. Reconstitution of the alpha1A subtype, but not the alpha1B, rescued alpha1ABKO myocytes from cell death induced by each stimulus. To address the mechanism, we examined alpha1-AR activation of extracellular signal-regulated kinase (ERK). In alpha1ABKO hearts, aortic constriction failed to activate ERK, and in alpha1ABKO myocytes, expression of a constitutively active MEK1 rescued alpha1ABKO myocytes from norepinephrine-induced death. In addition, only the alpha1A-AR activated ERK in alpha1ABKO myocytes, and expression of a dominant-negative MEK1 completely blocked alpha1A survival signaling in alpha1ABKO myocytes.
  
  
  CONCLUSIONS
  Our results demonstrate a direct protective effect of the alpha1A subtype in cardiac myocytes and define an alpha1A-ERK signaling pathway that is required for myocyte survival. Absence of the alpha1A-ERK pathway can explain the failure to activate ERK after aortic constriction in alpha1ABKO mice and can contribute to the development of apoptosis, dilated cardiomyopathy, and death.
authors.affiliation: Cardiovascular Research Institute, Sanford Research/USD, and Department of Medicine, The University of South Dakota School of Medicine, Sioux Falls, SD 57105, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Huang
authors.forename: Casey D
authors.initials: CD
authors.lastname: Wright
authors.forename: Chastity L
authors.initials: CL
authors.lastname: Merkwan
authors.forename: Nichole L
authors.initials: NL
authors.lastname: Baye
authors.forename: Qiangrong
authors.initials: Q
authors.lastname: Liang
authors.forename: Paul C
authors.initials: PC
authors.lastname: Simpson
authors.forename: Timothy D
authors.initials: TD
authors.lastname: O'Connell
mesh_terms: D000818:Animals; D017209:Apoptosis; D002311:Cardiomyopathy, Dilated / Q000209:etiology; D002470:Cell Survival; D002478:Cells, Cultured; D019610:Cytoprotection; D004317:Doxorubicin / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000502:physiology*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D048369:MAP Kinase Kinase 1 / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D032383:Myocytes, Cardiac / Q000166:cytology*; D009638:Norepinephrine / Q000494:pharmacology; D018340:Receptors, Adrenergic, alpha-1 / Q000145:classification / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D018340:Receptors, Adrenergic, alpha-1; D004317:Doxorubicin; D006861:Hydrogen Peroxide; D048049:Extracellular Signal-Regulated MAP Kinases; D048369:MAP Kinase Kinase 1; C482127:Map2k1 protein, mouse; D009638:Norepinephrine