Reduced force production during low blood flow to the heart correlates with altered troponin I phosphorylation.
A rat model of low myocardial blood flow was established to test the hypothesis that post-translational changes to proteins of the thin and thick muscle filaments correlate with decreased cardiac contractility. Following 3 days of low blood flow by constriction of the left anterior descending artery, rat hearts demonstrated a reduction in fractional shortening at rest and a relative decline in fractional shortening when challenged with high dose versus low dose dobutamine, reflecting reduced energy reserves. Permeabilized fibers from low blood flow hearts demonstrated a decline in maximum force per cross-section and Ca2+ sensitivity as compared to their sham operated counterparts. An examination of sarcomeric proteins by twodimensional gel electrophoresis, mass spectrometry, and phospho-specific antibodies provided evidence for Ser23/24 and Ser43/45 phosphorylation of troponin I (TnI). Total TnI phosphorylation was not different between the groups, but Ser23/24 phosphorylation declined with low blood flow, implying an accompanying increase in phosphorylation at other sites of TnI. Affinity chromatography demonstrated that TnI from low blood flow myocardium had reduced relative affinity to Ca2+ bound troponin C compared to TnI from sham operated hearts, providing a mechanism for reduced Ca2+ sensitivity of force production in low blood flow fibers. These findings suggest that altered TnI function, due to changes in the distribution of phosphorylated sites, is an early contributor to reduced contractility of the heart.
Christopher
Bridgette
B
Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, OH, 44106, USA.
Pizarro
Gresin O
GO
Nicholson
Bryson
B
Yuen
Samantha
S
Hoit
Brian D
BD
Ogut
Ozgur
O
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D001783:Blood Flow Velocity; D002118:Calcium / Q000378:metabolism*; D024722:Cardiac Myosins / Q000378:metabolism; D004280:Dobutamine / Q000494:pharmacology; D006321:Heart / Q000187:drug effects / Q000502:physiology*; D009200:Myocardial Contraction / Q000187:drug effects / Q000502:physiology*; D018994:Myosin Light Chains / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D019209:Troponin C / Q000378:metabolism; D019210:Troponin I / Q000378:metabolism*; D020107:Troponin T / Q000378:metabolism
D018994:Myosin Light Chains; D019209:Troponin C; D019210:Troponin I; D020107:Troponin T; C107184:myosin light chain 2; D004280:Dobutamine; D024722:Cardiac Myosins; D002118:Calcium