Underlying mechanism of the contractile dysfunction in atrophied ventricular myocytes from a murine model of hypothyroidism

Abstract

Hypothyroidism (Hypo) is a risk factor for cardiovascular diseases, including heart failure. Hypo rapidly induces Ca²⁺ mishandling and contractile dysfunction (CD), as well as atrophy and ventricular myocytes (VM) remodeling. Hypo decreases SERCA-to-phospholamban ratio (SERCA/PLB), and thereby contributes to CD. Nevertheless, detailed spatial and temporal Ca²⁺ cycling characterization in VM is missing, and contribution of other structural and functional changes to the mechanism underlying Ca²⁺ mishandling and CD, as transverse tubules (T-T) remodeling, mitochondrial density (D_mit) and energy availability, is unclear. Therefore, in a rat model of Hypo, we aimed to characterize systolic and diastolic Ca²⁺ signaling, T-T remodeling, D_mit, citrate synthase (CS) activity and high-energy phosphate metabolites (ATP and phosphocreatine). We confirmed a decrease in SERCA/PLB (59%), which slowed SERCA activity (48%), reduced SR Ca²⁺ (19%) and blunted Ca²⁺ transient amplitude (41%). Moreover, assessing the rate of SR Ca²⁺ release (dRel/dt), we found that early and maximum dRel/dt decreased, and this correlated with staggered Ca²⁺ transients. However, dRel/dt persisted during Ca²⁺ transient relaxation due to abundant late Ca²⁺ sparks. Isoproterenol significantly up-regulated systolic Ca²⁺ cycling. T-T were unchanged, hence, cannot explain staggered Ca²⁺ transients and altered dRel/dt. Therefore, we suggest that these might be caused by RyR2 clusters desynchronization, due to diminished Ca²⁺-dependent sensitivity of RyR2, which also caused a decrease in diastolic SR Ca²⁺ leak. Furthermore, D_mit was unchanged and CS activity slightly decreased (14%), however, the ratio phosphocreatine/ATP did not change, therefore, energy deficiency cannot account for Ca²⁺ and contractility dysregulation. We conclude that decreased SR Ca²⁺, due to slower SERCA, disrupts systolic RyR2 synchronization, and this underlies CD.

Keywords: Cardiac atrophy; Phospholamban; Ryanodine receptors type 2; SERCA; Transverse tubules.