SERCA mediated thermogenesis during contraction-relaxation cycle in rat skeletal muscle in vivo

Abstract

BACKGROUND: Across the lifespan, body temperature homeostasis is tightly regulated and impacts various physiological phenomena. In skeletal muscle, heat is produced via contraction-dependent (shivering) and independent (nonshivering) mechanisms, regulating the body temperature and muscle physiology including contractile properties. PURPOSE: In this study, we aimed to reveal the intracellular temperature dynamics in skeletal muscle in vivo during the contraction-relaxation cycle. Additionally, we tested the hypothesis that calcium ion (Ca2+) handling mediated by sarcoplasmic reticulum Ca2+ ATPase (SERCA) is involved in the thermogenesis process during contraction-relaxation cycle in skeletal muscle fibers. METHOD: The spinotrapezius muscle of anesthetized adult male Wistar rats (n = 18) was exteriorized and subjected to microinjection of fluorescent probe Cellular Thermoprobe for Fluorescence Ratio (49.3 μM) for intracellular temperature imaging in vivo. The fluorescence ratio (R: 580 nm / 515 nm) was measured in vivo during temperature increases induced by an external heater for thermoprobe calibration. R was also measured following Ca2+ injection (3.9 nL, 2.0 mM), and Ca2+ injection with SERCA inhibition by cyclopiazonic acid (CPA, 100 μM). RESULTS: The fluorescence ratio linearly increased with the increase of the muscle surface temperature from 25 °C to 40 °C (r2 = 0.97, P < 0.01). The intracellular temperature in muscle was increased and sustained following the Ca2+ injection and this thermogenetic response was significantly suppressed with SERCA inhibition (Ca2+: 38.3 ± 1.4 °C vs Ca2++CPA: 28.3 ± 2.8 °C, P < 0.01 at 1 min following injection). Importantly, elevated muscle temperature occurred predominantly in the muscle relaxation phase that followed the muscle shortening: That shortening occurred immediately and transiently after the Ca2+ injection. CONCLUSION: In this investigation, we demonstrated a novel technique for in vivo intracellular temperature imaging of skeletal muscle. Herein we demonstrate that substantial heat is generated concomitantly with SERCA mediated Ca2+ handling and muscle relaxation. This work was supported by JSPS KAKENHI Grant Numbers 20H04074, 19K22800. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.