Phosphorylation in coronary artery cold-induced contraction in the newborn lamb.
Myocardial dysfunction after hypothermic protection has been linked to various mechanisms. Coronary vasospasm in particular may be responsible for ischemic injury during reperfusion. Herein we hypothesized that coronary arteries (CA) sustain a cold-induced contraction during hypothermia mediated by a protein tyrosine kinase (PTK)-/protein tyrosine phosphatase (PTP)-dependent pathway. Isolated newborn lamb CA rings were studied in a tissue bath for isometric contraction during 2-h profound (17 degrees C) or ultra-profound (7 degrees C) hypothermia. In parallel, protein tyrosine phosphorylation was evaluated by use of the Western blot technique. Na-orthovanadate (SOV) and genestein (GEN) were used separately and in combination to evaluate the effect of PTK/PTP activation on CA contraction and tyrosine phosphorylation during cooling (17 or 7 degrees C) vs 37 degrees C. Cooling from 37 to 7 degrees C induced transient contraction at approximately 17 degrees C (29% KCl response), which was more prominent during rewarming to 37 degrees C (36% KCl). Cooling to 17 degrees C resulted in sustained contraction (7-10% KCl), which was reversible upon rewarming. Cold-induced contraction was significantly enhanced by SOV (7- to 10-fold at 17 degrees C; 2-fold at 7 degrees C) and abolished by GEN. Concurrently, tyrosine phosphorylation of 33-, 45-, and 104-kDa proteins increased during cooling (35-100% at 17 degrees C; 46-66% at 7 degrees C). Tyrosine phosphorylation was similarly enhanced by SOV (1.7- to 2.3-fold at 17 degrees C; 2.9- to 3.9-fold at 7 degrees C) and abolished by GEN in the presence or absence of SOV. These results support a prominent role for the PTK/PTP signal transduction pathway in the coronary artery cold-induced contraction. This information provides one possible biomolecular mechanism linked to ischemia/reperfusion pathophysiology of CA in neonatal hearts exposed to hypothermic myocardial protection.
Dahdah
N S
NS
Department of Pediatrics, MetroHealth Medical Center, Cleveland, Ohio 44109, USA.
Russo
P
P
Wagerle
L C
LC
D000818:Animals; D000831:Animals, Newborn; D003080:Cold Temperature; D003329:Coronary Vasospasm / Q000209:etiology / Q000503:physiopathology; D003331:Coronary Vessels / Q000378:metabolism / Q000503:physiopathology*; D007036:Hypothermia, Induced / Q000009:adverse effects*; D066298:In Vitro Techniques; D015428:Myocardial Reperfusion Injury / Q000209:etiology / Q000503:physiopathology; D009926:Organ Preservation / Q000009:adverse effects; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D012756:Sheep; D015398:Signal Transduction; D014661:Vasoconstriction / Q000502:physiology*
D011505:Protein-Tyrosine Kinases; D017027:Protein Tyrosine Phosphatases