Action potential–driven Ca2+ currents from the transverse tubules (t-tubules) trigger synchronous Ca2+ release from the sarcoplasmic reticulum of cardiomyocytes. Loss of t-tubules has been reported in cardiac diseases, including heart failure, but the effect of uncoupling t-tubules from the sarcolemma on cardiac muscle mechanics remains largely unknown. We dissected intact rat right ventricular trabeculae and compared force, sarcomere length, and intracellular Ca2+ in control trabeculae with trabeculae in which the t-tubules were uncoupled from the plasma membrane by formamide-induced osmotic shock (detubulation). We verified disconnection of a consistent fraction of t-tubules from the sarcolemma by two-photon fluorescence imaging of FM4-64–labeled membranes and by the absence of tubular action potential, which was recorded by random access multiphoton microscopy in combination with a voltage-sensitive dye (Di-4-AN(F)EPPTEA). Detubulation reduced the amplitude and prolonged the duration of Ca2+ transients, leading to slower kinetics of force generation and relaxation and reduced twitch tension (1 Hz, 30°C, 1.5 mM [Ca2+]o). No mechanical changes were observed in rat left atrial trabeculae after formamide shock, consistent with the lack of t-tubules in rodent atrial myocytes. Detubulation diminished the rate-dependent increase of Ca2+-transient amplitude and twitch force. However, maximal twitch tension at high [Ca2+]o or in post-rest potentiated beats was unaffected, although contraction kinetics were slower. The ryanodine receptor (RyR)2 Ca-sensitizing agent caffeine (200 μM), which increases the velocity of transverse Ca2+ release propagation in detubulated cardiomyocytes, rescued the depressed contractile force and the slower twitch kinetics of detubulated trabeculae, with negligible effects in controls. We conclude that partial loss of t-tubules leads to myocardial contractile abnormalities that can be rescued by enhancing and accelerating the propagation of Ca2+- induced Ca2+ release to orphan RyR2 clusters.

Impact of detubulation on force and kinetics of cardiac muscle contraction / Ferrantini C; Coppini R; Sacconi L; Tosi B; Zhang ML; Wang GL; de Vries E; Hoppenbrouwers E; Pavone F; Cerbai E; Tesi C; Poggesi C; Ter Keurs HE. - In: JOURNAL OF GENERAL PHYSIOLOGY. - ISSN 0022-1295. - STAMPA. - 143:(2014), pp. 783-797.

Impact of detubulation on force and kinetics of cardiac muscle contraction

FERRANTINI, CECILIA;COPPINI, RAFFAELE;SACCONI, LEONARDO;TOSI, BENEDETTA;PAVONE, FRANCESCO SAVERIO;CERBAI, ELISABETTA;TESI, CHIARA;POGGESI, CORRADO;
2014

Abstract

Action potential–driven Ca2+ currents from the transverse tubules (t-tubules) trigger synchronous Ca2+ release from the sarcoplasmic reticulum of cardiomyocytes. Loss of t-tubules has been reported in cardiac diseases, including heart failure, but the effect of uncoupling t-tubules from the sarcolemma on cardiac muscle mechanics remains largely unknown. We dissected intact rat right ventricular trabeculae and compared force, sarcomere length, and intracellular Ca2+ in control trabeculae with trabeculae in which the t-tubules were uncoupled from the plasma membrane by formamide-induced osmotic shock (detubulation). We verified disconnection of a consistent fraction of t-tubules from the sarcolemma by two-photon fluorescence imaging of FM4-64–labeled membranes and by the absence of tubular action potential, which was recorded by random access multiphoton microscopy in combination with a voltage-sensitive dye (Di-4-AN(F)EPPTEA). Detubulation reduced the amplitude and prolonged the duration of Ca2+ transients, leading to slower kinetics of force generation and relaxation and reduced twitch tension (1 Hz, 30°C, 1.5 mM [Ca2+]o). No mechanical changes were observed in rat left atrial trabeculae after formamide shock, consistent with the lack of t-tubules in rodent atrial myocytes. Detubulation diminished the rate-dependent increase of Ca2+-transient amplitude and twitch force. However, maximal twitch tension at high [Ca2+]o or in post-rest potentiated beats was unaffected, although contraction kinetics were slower. The ryanodine receptor (RyR)2 Ca-sensitizing agent caffeine (200 μM), which increases the velocity of transverse Ca2+ release propagation in detubulated cardiomyocytes, rescued the depressed contractile force and the slower twitch kinetics of detubulated trabeculae, with negligible effects in controls. We conclude that partial loss of t-tubules leads to myocardial contractile abnormalities that can be rescued by enhancing and accelerating the propagation of Ca2+- induced Ca2+ release to orphan RyR2 clusters.
2014
143
783
797
Ferrantini C; Coppini R; Sacconi L; Tosi B; Zhang ML; Wang GL; de Vries E; Hoppenbrouwers E; Pavone F; Cerbai E; Tesi C; Poggesi C; Ter Keurs HE...espandi
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/872838
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