Work-loop contractions reveal that the afterload-dependent time course of cardiac Ca²⁺ transients is modulated by...

Preload and afterload dictate the dynamics of the cyclical work-loop contraction that the heart undergoes in vivo. Cellular Ca²⁺ dynamics drive contraction, but the effects of afterload alone on the Ca²⁺ transient are inconclusive. To our knowledge, no study has investigated whether the putative afterload dependence of the Ca²⁺ transient is preload dependent. This study is designed to provide the first insight into the Ca²⁺ handling of cardiac trabeculae undergoing work-loop contractions, with the aim to examine whether the conflicting afterload dependency of the Ca²⁺ transient can be accounted for by considering preload under isometric and physiological work-loop contractions. Thus, we subjected ex vivo rat right-ventricular trabeculae, loaded with the fluorescent dye Fura-2, to work-loop contractions over a wide range of afterloads at two preloads while measuring stress, length changes, and Ca²⁺ transients. Work-loop control was implemented with a real-time Windkessel model to mimic the contraction patterns of the heart in vivo. We extracted a range of metrics from the measured steady-state twitch stress and Ca²⁺ transients, including the amplitudes, time courses, rates of rise, and integrals. Results show that parameters of stress were afterload and preload dependent. In contrast, the parameters associated with Ca²⁺ transients displayed a mixed dependence on afterload and preload. Most notably, its time course was afterload dependent, an effect augmented at the greater preload. This study reveals that the afterload dependence of cardiac Ca²⁺ transients is modulated by preload, which brings the study of Ca²⁺ transients during isometric contractions into question when aiming to understand physiological Ca²⁺ handling.NEW & NOTEWORTHY This study is the first examination of Ca²⁺ handling in trabeculae undergoing work-loop contractions. These data reveal that reducing preload diminishes the influence of afterload on the decay phase of the cardiac Ca²⁺ transient. This is significant as it reconciles inconsistencies in the literature regarding the influence of external loads on cardiac Ca²⁺ handling. Furthermore, these findings highlight discrepancies between Ca²⁺ handling during isometric and work-loop contractions in cardiac trabeculae operating at their optimal length.