Search

A regular heartbeat is essential for maintaining the homeostasis of the vertebrate body. However, environmental pollutants, oxygen deficiency and extreme temperatures can impair heart function in fish. In this Review, we provide an integrative view of the molecular origins of cardiac arrhythmias and their functional consequences, from the level of ion channels to cardiac electrical activity in living fish. First, we describe the current knowledge of the cardiac excitation-contraction coupling of fish, as the electrical activity of the heart and intracellular Ca2+ regulation act as a platform for cardiac arrhythmias. Then, we compile findings on cardiac arrhythmias in fish. Although fish can experience several types of cardiac arrhythmia under stressful conditions, the most typical arrhythmia in fish - both under heat stress and in the presence of toxic substances - is atrioventricular block, which is the inability of the action potential to progress from the atrium to the ventricle. Early and delayed afterdepolarizations are less common in fish hearts than in the hearts of endotherms, perhaps owing to the excitation-contraction coupling properties of the fish heart. In fish hearts, Ca2+-induced Ca2+ release from the sarcoplasmic reticulum plays a smaller role than Ca2+ influx through the sarcolemma. Environmental changes and ion channel toxins can induce arrhythmias in fish and weaken their tolerance to environmental stresses. Although different from endotherm hearts in many respects, fish hearts can serve as a translational model for studying human cardiac arrhythmias, especially for human neonates., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Heat tolerance of heart rate in fish is suggested to be limited by impaired electrical excitation of the ventricle due to the antagonistic effects of high temperature on Na+ (INa) and K+ (IK1) ion currents (INa is depressed at high temperatures while IK1 is resistant to them). To examine the role of Na+ channel proteins in heat tolerance of INa, we compared temperature dependencies of zebrafish (Danio rerio, warm-dwelling subtropical species) and rainbow trout (Oncorhynchus mykiss, cold-active temperate species) ventricular INa, and INa generated by the cloned zebrafish and rainbow trout NaV1.4 and NaV1.5 Na+ channels in human embryonic kidney (HEK) cells. Whole-cell patch-clamp recordings showed that zebrafish ventricular INa has better heat tolerance and slower inactivation kinetics than rainbow trout ventricular INa. In contrast, heat tolerance and inactivation kinetics of zebrafish and rainbow trout NaV1.4 channels are similar when expressed in the identical cellular environment of HEK cells. The same applies to NaV1.5 channels. These findings indicate that thermal adaptation of ventricular INa is largely achieved by differential expression of Na+ channel alpha subunits: zebrafish that tolerate higher temperatures mainly express the slower NaV1.5 isoform, while rainbow trout that prefer cold waters mainly express the faster NaV1.4 isoform. Differences in elasticity (stiffness) of the lipid bilayer and/or accessory protein subunits of the channel assembly may also be involved in thermal adaptation of INa. The results are consistent with the hypothesis that slow Na+ channel kinetics are associated with increased heat tolerance of cardiac excitation., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2021. Published by The Company of Biologists Ltd.)

At critically high temperature, cardiac output in fish collapses as a result of depression of heart rate (bradycardia). However, the cause of bradycardia remains unresolved. To investigate this, rainbow trout ( Oncorhynchus mykiss ; acclimated at 12°C) were exposed to acute warming while electrocardiograms were recorded. From 12°C to 25.3°C, electrical excitation between different parts of the heart was coordinated, but above 25.3°C, atrial and ventricular beating rates became partly dissociated because of 2:1 atrioventricular (AV) block. With further warming, atrial rate increased to a peak value of 188±22 beats min -1 at 27°C, whereas the ventricle rate peaked at 124±10 beats min -1 at 25.3°C and thereafter dropped to 111±15 beats min -1 at 27°C. In single ventricular myocytes, warming from 12°C to 25°C attenuated electrical excitability as evidenced by increases in rheobase current and the size of critical depolarization required to trigger action potential. Depression of excitability was caused by temperature-induced decrease in input resistance (sarcolemmal K + leak via the outward I K1 current) of resting myocytes and decrease in inward charge transfer by the Na + current ( I Na ) of active myocytes. Collectively, these findings show that at critically high temperatures AV block causes ventricular bradycardia owing to the increased excitation threshold of the ventricle, which is due to changes in the passive (resting ion leak) and active (inward charge movement) electrical properties of ventricular myocytes. The sequence of events from the level of ion channels to cardiac function in vivo provides a mechanistic explanation for the depression of cardiac output in fish at critically high temperature., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

Electrical excitability (EE) is vital for cardiac function and strongly modulated by temperature and external K + concentration ([K + ] o ), as formulated in the hypothesis of temperature-dependent deterioration of electrical excitability (TDEE). As little is known about EE of arctic stenothermic fishes, we tested the TDEE hypothesis on ventricular myocytes of polar cod ( Boreogadus saida ) and navaga ( Eleginus nawaga ) of the Arctic Ocean and those of temperate freshwater burbot ( Lota lota ). Ventricular action potentials (APs) were elicited in current-clamp experiments at 3, 9 and 15°C, and AP characteristics and the current needed to elicit APs were examined. At 3°C, ventricular APs of polar cod and navaga were similar but differed from those of burbot in having a lower rate of AP upstroke and a higher rate of repolarization. EE of ventricular myocytes - defined as the ease with which all-or-none APs are triggered - was little affected by acute temperature changes between 3 and 15°C in any species. However, AP duration (APD 50 ) was drastically reduced at higher temperatures. Elevation of [K + ] o from 3 to 5.4 mmol l -1 and further to 8 mmol l -1 at 3, 9 and 15°C strongly affected EE and AP characteristics in polar cod and navaga, but had a lesser effect in burbot. In all species, ventricular excitation was resistant to acute temperature elevations, while small increases in [K + ] o severely compromised EE, in particular in the marine stenotherms. This suggests that EE of the heart in these Gadiformes species is resistant against acute warming, but less so against the simultaneous temperature and exercise stresses., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Calcium channels are necessary for cardiac excitation-contraction (E-C) coupling, but Ca 2+ channel composition of fish hearts is still largely unknown. To this end, we determined transcript expression of Ca 2+ channels in the heart of zebrafish ( Danio rerio ), a popular model species. Altogether, 18 Ca 2+ channel α-subunit genes were expressed in both atrium and ventricle. Transcripts for 7 L-type (Ca v 1.1a, Ca v 1.1b, Ca v 1.2, Ca v 1.3a, Ca v 1.3b, Ca v 1.4a, Ca v 1.4b), 5 T-type (Ca v 3.1, Ca v 3.2a, Ca v 3.2b, Ca v 3.3a, Ca v 3.3b) and 6 P/Q-, N- and R-type (Ca v 2.1a, Ca v 2.1b, Ca v 2.2a, Ca v 2.2b, Ca v 2.3a, Ca v 2.3b) Ca 2+ channels were expressed. In the ventricle, T-type channels formed 54.9%, L-type channels 41.1% and P/Q-, N- and R-type channels 4.0% of the Ca 2+ channel transcripts. In the atrium, the relative expression of T-type and L-type Ca 2+ channel transcripts was 64.1% and 33.8%, respectively (others accounted for 2.1%). Thus, at the transcript level, T-type Ca 2+ channels are prevalent in zebrafish atrium and ventricle. At the functional level, peak densities of ventricular T-type ( I CaT ) and L-type ( I CaL ) Ca 2+ current were 6.3±0.8 and 7.7±0.8 pA pF -1 , respectively. I CaT mediated a sizeable sarcolemmal Ca 2+ influx into ventricular myocytes: the increment in total cellular Ca 2+ content via I CaT was 41.2±7.3 µmol l -1 , which was 31.7% of the combined Ca 2+ influx (129 µmol l -1 ) via I CaT and I CaL (88.5±20.5 µmol l -1 ). The diversity of expressed Ca 2+ channel genes in zebrafish heart is high, but dominated by the members of the T-type subfamily. The large ventricular I CaT is likely to play a significant role in E-C coupling., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2018. Published by The Company of Biologists Ltd.)

The effects of sustained anoxia on cardiac electrical excitability were examined in the anoxia-tolerant crucian carp (Carassius carassius). The electrocardiogram (ECG) and expression of excitation-contraction coupling genes were studied in fish acclimatised to normoxia in summer (+18°C) or winter (+2°C), and in winter fish after 1, 3 and 6 weeks of anoxia. Anoxia induced a sustained bradycardia from a heart rate of 10.3±0.77 beats min -1 to 4.1±0.29 beats min -1 (P<0.05) after 5 weeks, and heart rate slowly recovered to control levels when oxygen was restored. Heart rate variability greatly increased under anoxia, and completely recovered under re-oxygenation. The RT interval increased from 2.8±0.34 s in normoxia to 5.8±0.44 s under anoxia (P<0.05), which reflects a doubling of the ventricular action potential (AP) duration. Acclimatisation to winter induced extensive changes in gene expression relative to summer-acclimatised fish, including depression in those genes coding for the sarcoplasmic reticulum calcium pump (Serca2a&#95;q2) and ATP-sensitive K + channels (Kir6.2) (P<0.05). Genes of delayed rectifier K + (kcnh6) and Ca 2+ channels (cacna1c) were up-regulated in winter fish (P<0.05). In contrast, the additional challenge of anoxia caused only minor changes in gene expression, e.g. depressed expression of Kir2.2b K + channel gene (kcnj12b), whereas expression of Ca 2+ (cacna1a, cacna1c and cacna1g) and Na + channel genes (scn4a and scn5a) was not affected. These data suggest that low temperature pre-conditions the crucian carp heart for winter anoxia, whereas sustained anoxic bradycardia and prolongation of AP duration are directly induced by oxygen shortage without major changes in gene expression., (© 2017. Published by The Company of Biologists Ltd.)

The upper thermal tolerance and mechanisms of heat-induced cardiac failure in the brown trout (Salmo trutta fario) was examined. The point above which ion channel function and sinoatrial contractility in vitro, and electrocardiogram (ECG) in vivo, started to fail (break point temperature, BPT) was determined by acute temperature increases. In general, electrical excitation of the heart was most sensitive to heat in the intact animal (electrocardiogram, ECG) and least sensitive in isolated cardiac myocytes (ion currents). BPTs of Ca(2+) and K(+) currents of cardiac myocytes were much higher (>28°C) than BPT of in vivo heart rate (23.5 ± 0.6°C) (P<0.05). A striking exception among sarcolemmal ion conductances was the Na(+) current (INa), which was the most heat-sensitive molecular function, with a BPT of 20.9 ± 0.5°C. The low heat tolerance of INa was reflected as a low BPT for the rate of action potential upstroke in vitro (21.7 ± 1.2°C) and the velocity of impulse transmission in vivo (21.9 ± 2.2°C). These findings from different levels of biological organization strongly suggest that heat-dependent deterioration of Na(+) channel function disturbs normal spread of electrical excitation over the heart, leading to progressive variability of cardiac rhythmicity (missed beats, bursts of fast beating), reduction of heart rate and finally cessation of the normal heartbeat. Among the cardiac ion currents INa is 'the weakest link' and possibly a limiting factor for upper thermal tolerance of electrical excitation in the brown trout heart. Heat sensitivity of INa may result from functional requirements for very high flux rates and fast gating kinetics of the Na(+) channels, i.e. a trade-off between high catalytic activity and thermal stability.

The present study examines the importance of the Na+ current (INa) in the excitability of atrial and ventricular myocardium of the rainbow trout heart. Whole-cell patch-clamp under reduced sarcolemmal Na+ gradient showed that the density of INa is similar in atrial and ventricular myocytes of the trout heart, and the same result was obtained when INa was elicited by chamber-specific action potentials (AP) in normal physiological saline solution. However, the maximum rate (Vmax) of AP upstroke, measured with microelectrodes in intact trout heart, was 21% larger in atrium than ventricle, and thus in variance with the similar INa density of the two myocyte types. Furthermore, Vmax calculated from the INa was 2.1 and 3.2 times larger for atrium and ventricle, respectively, than the values obtained from the APs. The discrepancy between INa of isolated myocytes and Vmax of intact muscle is only partly explained by the inward rectifier K+ current (IK1), which overlaps INa and decreases the net depolarising current. Clear differences exist in the voltage dependence of steady-state activation and inactivation as well as in the inactivation kinetics of INa between atrial and ventricular myocytes. As a result of a more negative voltage dependence of INa activation, smaller IK1 and higher input resistance of atrial myocytes, the voltage threshold for AP generation is more negative in atrium than ventricle of the trout heart. These findings suggest that atrial muscle is more readily excitable than ventricular muscle, and this difference is partly due to the properties of the atrial INa.

The present study was designed to test the hypothesis that temperature acclimation modifies sarcolemmal Na+ current (INa) of the fish cardiac myocytes differently depending on the animal's lifestyle in the cold. Two eurythermal fish species with different physiological strategies for surviving in the cold, a cold-dormant crucian carp (Carassius carassius L.) and a cold-active rainbow trout (Oncorhynchus mykiss), were used in acclimation experiments. The INa of carp and trout were also compared with INa of a cold stenothermal burbot (Lota lota). In accordance with the hypothesis, cold-acclimation decreased the density of INa in crucian carp and increased it in rainbow trout, suggesting depression of impulse conduction in cold-acclimated carp and positive compensation of impulse propagation in cold-acclimated trout. The steady-state activation curve of trout INa was shifted by 6 mV to more negative voltages by cold acclimation, which probably lowers the stimulus threshold for action potentials and further improves cardiac excitability in the cold. In burbot myocytes, the INa density was high and the position of the steady-state activation curve on the voltage axis was even more negative than in trout or carp myocytes, suggesting that the burbot INa is adapted to maintain high excitability and conductivity in the cold. The INa of the burbot heart differed from those of carp and trout in causing four times larger charge influx per excitation, which suggests that INa may also have a significant role in cardiac excitation-contraction coupling of the burbot heart. In summary, INa of fish cardiac myocytes shows thermal plasticity that is different in several respects in cold-dormant and cold-active species and thus has a physiologically meaningful role in supporting the variable life styles and habitat conditions of each species.