%A Woodward,Mike %A Previs,Michael %A Mader,Timothy %A Debold,Edward %D 2015 %J Frontiers in Physiology %C %F %G English %K Cardiac arrest,Resuscitation,Myosin,Troponin,Phosphorylation,motility %Q %R 10.3389/fphys.2015.00199 %W %L %M %P %7 %8 2015-July-16 %9 Original Research %+ Dr Edward Debold,University of Massachusetts, Amherst,Kinesiology,158 Totman Bldg.,Amherst,01002,MA,United States,edebold@kin.umass.edu %+ Dr Edward Debold,University of Massachusetts, Amherst,Molecular and Cellular Biology,Amherst,01002,MA,United States,edebold@kin.umass.edu %# %! Molecular effects of cardiac arrest %* %< %T Modifications of myofilament protein phosphorylation and function in response to cardiac arrest induced in a swine model %U https://www.frontiersin.org/articles/10.3389/fphys.2015.00199 %V 6 %0 JOURNAL ARTICLE %@ 1664-042X %X Cardiac arrest is a prevalent condition with a poor prognosis, attributable in part to persistent myocardial dysfunction following resuscitation. The molecular basis of this dysfunction remains unclear. We induced cardiac arrest in a porcine model of acute sudden death and assessed the impact of ischemia and reperfusion on the molecular function of isolated cardiac contractile proteins. Cardiac arrest was electrically induced, left untreated for 12 min, and followed by a resuscitation protocol. With successful resuscitations, the heart was reperfused for 2 h (IR2) and the muscle harvested. In failed resuscitations, tissue samples were taken following the failed efforts (IDNR). Actin filament velocity, using myosin isolated from IR2 or IDNR cardiac tissue, was nearly identical to myosin from the control tissue in a motility assay. However, both maximal velocity (25% faster than control) and calcium sensitivity (pCa50 6.57 ± 0.04 IDNR vs. 6.34 ± 0.07 control) were significantly (p < 0.05) enhanced using native thin filaments (actin+troponin+tropomyosin) from IDNR samples, suggesting that the enhanced velocity is mediated through an alteration in muscle regulatory proteins (troponin+tropomyosin). Mass spectrometry analysis showed that only samples from the IR2 had an increase in total phosphorylation levels of troponin (Tn) and tropomyosin (Tm), but both IR2 and IDNR samples demonstrated a significant shift from mono-phosphorylated to bis-phosphorylated forms of the inhibitory subunit of Tn (TnI) compared to control. This suggests that the shift to bis-phosphorylation of TnI is associated with the enhanced function in IDNR, but this effect may be attenuated when phosphorylation of Tm is increased in tandem, as observed for IR2. There are likely many other molecular changes induced following cardiac arrest, but to our knowledge, these data provide the first evidence that this form cardiac arrest can alter the in vitro function of the cardiac contractile proteins.