Nuclear receptors PPARbeta/delta and PPARalpha direct distinct metabolic regulatory programs in the mouse heart.

In the diabetic heart, chronic activation of the PPARalpha pathway drives excessive fatty acid (FA) oxidation, lipid accumulation, reduced glucose utilization, and cardiomyopathy. The related nuclear receptor, PPARbeta/delta, is also highly expressed in the heart, yet its function has not been fully delineated. To address its role in myocardial metabolism, we generated transgenic mice with cardiac-specific expression of PPARbeta/delta, driven by the myosin heavy chain (MHC-PPARbeta/delta mice).

Molecular and integrated biology of thin filament protein phosphorylation in heart muscle.

An increasing body of evidence points to posttranslational modifications of the thin filament regulatory proteins, cardiac troponin T (cTnT) and cardiac troponin I (cTnI) by protein kinase C (PKC) phosphorylation as important in both long- and short-term regulation of cardiac function and potentially implicated in the transition between compensated hypertrophy and decompensation. The main sites for PKC-dependent phosphorylation on cTnI are Ser43, Ser45, and Thr144 and on cTnT are Thr197, Ser201, Thr206, and Thr287 (mouse sequence). We analyzed the function of each phosphorylation residue using a phosphorylation mimic approach introducing glutamates (E) at PKC phosphorylation sites and then measuring the isometric tension of fiber bundles exchanged with these mutants.

Effects of protein kinase C dependent phosphorylation and a familial hypertrophic cardiomyopathy-related mutation of cardiac troponin I on structural transition of troponin C and myofilament activation.

In experiments reported here, we compared tension and thin filament Ca(2+) signaling in preparations containing either wild-type cardiac troponin I (cTnI) or a mutant cTnI with an R146G mutation [cTnI(146G)] linked to familial hypertrophic cardiomyopathy. Myofilament function is altered in association with cTnI phosphorylation by protein kinase C (PKC), which is activated in hypertrophy. Whether there are differential effects of PKC phosphorylation on cTnI compared to cTnI(146G) remains unknown.

Altered signaling surrounding the C-lobe of cardiac troponin C in myofilaments containing an alpha-tropomyosin mutation linked to familial hypertrophic cardiomyopathy.

A region of interaction between the near N-terminal of cardiac troponin I (cTnI) and the C-lobe of troponin C (cTnC), where troponin T (cTnT) binds, appears to be critical in regulation of myofilament Ca(2+)-activation. We probed whether functional consequences of modulation of this interface influence the function of tropomyosin (Tm) in thin filament activation. We modified the C-lobe of cTnC directly by addition of the Ca(2+)-sensitizer, EMD 57033, and indirectly by replacing native cTnI with cTnI-containing Glu residues at Ser-43 and Ser-45 (cTnI-S43E/S45E) in myofilaments from hearts of non-transgenic (NTG) and transgenic (TG) mice expressing a point mutation on alpha-Tm (E180G) linked to familial hypertrophic cardiomyopathy.

Phosphorylation or glutamic acid substitution at protein kinase C sites on cardiac troponin I differentially depress myofilament tension and shortening velocity.

There is evidence that multi-site phosphorylation of cardiac troponin I (cTnI) by protein kinase C is important in both long- and short-term regulation of cardiac function. To determine the specific functional effects of these phosphorylation sites (Ser-43, Ser-45, and Thr-144), we measured tension and sliding speed of thin filaments in reconstituted preparations in which endogenous cTnI was replaced with cTnI phosphorylated by protein kinase C-epsilon or mutated to cTnI-S43E/S45E/T144E, cTnI-S43E/S45E, or cTnI-T144E. We used detergent-skinned mouse cardiac fiber bundles to measure changes in Ca(2+)-dependence of force.

Integration of pathways that signal cardiac growth with modulation of myofilament activity.

We review evidence for integrated effects of the signals that promote cardiac growth and remodeling and that modify the processes of excitation-contraction coupling. We have focused on integration of alterations in myofilament function with cell growth on the basis of genetic linkage analysis demonstrating that sarcomeric mutations are causal in hypertrophic cardiomyopathies. This evidence argues strongly for a path of communication between the intrinsic functional changes associated with a sarcomeric protein mutation and nuclear events.