Force-frequency Relationship Research Articles

The reduced myofilament responsiveness to calcium contributes to the negative force-frequency relationship in rat cardiomyocytes: role of reactive oxygen species and p-38 map kinase.

The force-frequency relationship (FFR) is an important intrinsic regulatory mechanism of cardiac contractility. However, a decrease (negative FFR) or no effect (flat FFR) on contractile force in response to an elevation of heart rate is present in the normal rat or in human heart failure. Reactive oxygen species (ROS) can act as intracellular signaling molecules activating diverse kinases as calcium-calmodulin-dependent protein kinase II (CaMKII) and p-38 MAP kinase (p-38K). Our aim was to elucidate the intracellular molecules implicated in the FFR of isolated rat ventricular myocytes. The myocytes were field-stimulated via two-platinum electrodes. Sarcomere length was recorded with a video camera. Ca2+ transients and intracellular pHi were recorded by epifluorescence. Increasing frequency from 0.5 to 3Hz decreased cell shortening without changes in pHi. This negative FFR was changed to positive FFR when the myocytes were pre-incubated with the ROS scavenger MPG, the NADPH oxidase blocker apocynin, or by inhibiting mitochondrial ROS production with 5-HD. Similar results were obtained when the cells were pre-incubated with the CaMKII blocker, KN-93, or the p-38K inhibitor, SB-202190. Consistently, the levels of phosphorylation of p-38K and the oxidation of CaMKII were significantly higher at 2Hz than at 0.5Hz. Despite the presence of positive inotropic effect during stimulation frequency enhancement, Ca2+ transient amplitudes were reduced in MPG- and SB-202190-treated myocytes. In conclusion, our results indicate that the activation of the intracellular pathway involving ROS-CaMKII-p-38K contributes to the negative FFR of rat cardiomyocytes, likely by desensitizing the response of contractile myofilaments to Ca2+.

Measuring Neuromuscular Junction Functionality.

Neuromuscular junction (NMJ) functionality plays a pivotal role when studying diseases in which the communication between motor neuron and muscle is impaired, such as aging and amyotrophic lateral sclerosis (ALS). Here we describe an experimental protocol that can be used to measure NMJ functionality by combining two types of electrical stimulation: direct muscle membrane stimulation and the stimulation through the nerve. The comparison of the muscle response to these two different stimulations can help to define, at the functional level, potential alterations in the NMJ that lead to functional decline in muscle. Ex vivo preparations are suited to well-controlled studies. Here we describe an intensive protocol to measure several parameters of muscle and NMJ functionality for the soleus-sciatic nerve preparation and for the diaphragm-phrenic nerve preparation. The protocol lasts approximately 60 min and is conducted uninterruptedly by means of a custom-made software that measures the twitch kinetics properties, the force-frequency relationship for both muscle and nerve stimulations, and two parameters specific to NMJ functionality, i.e. neurotransmission failure and intratetanic fatigue. This methodology was used to detect damages in soleus and diaphragm muscle-nerve preparations by using SOD1G93A transgenic mouse, an experimental model of ALS that ubiquitously overexpresses the mutant antioxidant enzyme superoxide dismutase 1 (SOD1).

Strain Rate in Children and Young Piglets Mirrors Changes in Contractility and Demonstrates a Force-Frequency Relationship

In adult humans and pig models, strain rate (SR) correlates strongly with invasive measures of contractility but does not demonstrate a force-frequency relationship, which is a fundamental behavior of myocardial contraction. Given the considerable maturational changes of the myocardium from fetal stages to adulthood, extrapolation of adult findings to the young heart may not be appropriate. We sought to evaluate the SR response of the immature heart to increased heart rate (HR) and inotropic stimulation. The study consisted of two parts. In part 1, children without obvious structural or functional cardiac abnormalities were evaluated following successful radiofrequency ablation. Echocardiography was performed at baseline HR and then with atrial pacing and isoprenaline infusion titrated to achieve 130% of baseline HR. Left ventricular (LV) speckle tracking-derived SR and tissue Doppler isovolumic acceleration (IVA, a load-independent marker of contractility) were measured. In part 2, young piglets were submitted to atrial pacing at 200, 230, and 260 bpm. Invasive LV dP/dt was assessed, and speckle tracking-derived SR was measured at all stages. Repeated measures analysis of variance was used for comparison with baseline values. In part 1, the LV SR and IVA (septal and lateral) in 23 children (ages 7.8-17.5years) increased significantly with pacing and isoprenaline infusion (P<.001). In part 2, SR and invasive dP/dt increased significantly with increasing HR in young piglets (1-17days; P<.01 and P<.001, respectively). Both LV SR and dP/dt plateaued at highest HRs concurrent with the decrease in LV end-diastolic dimension from baseline (73.0% ±9.9% of the baseline value at 260 bpm; P<.001). SR in children is augmented with chronotropic and inotropic stimulation and in young infant piglets with chronotropic stimulation; in both children and piglets it has a force-frequency relationship, abehavior that is consistent with a measure of contractility.

A simple laboratory exercise with rat isolated esophagus and stomach fundus to reveal functional differences between striated and smooth muscle cells.

This study describes an undergraduate student laboratory activity using isolated preparations from rat gastrointestinal tissues that possess contractile profiles typically exhibited by striated and smooth muscle cells. While students are introduced to an ex vivo methodology, they can compare differences in trace experiments, twitch aspects, phasic and tonic properties, force-frequency relationships, and pharmacological responsiveness of esophageal (striated) and fundic (smooth muscle) segments. Muscle strips were subjected to electrical field stimulation (EFS) applied by platinum electrodes immersed in the physiological solution. The contractile profile of EFS responses varied between these two types of gut preparations. Atropine and tubocurarine revealed differential inhibitory influences in esophagus or fundus tissues; caffeine and procaine produced similar effects, i.e., potentiation and blockade of the EFS-induced contractile response in these tissues, respectively. Experimental results obtained during the activity helped the improvement of student learning about basic concepts previously discussed in theoretical lectures. To measure student learning with this laboratory exercise, a questionnaire was applied before and after the activity, and the number of expected correct answers, concerning the mechanisms of contraction in striated and smooth muscle, could be clearly evidenced.

Impaired cardiac output during exercise in adults operated for ventricular septal defect in childhood: a hitherto unrecognised pathophysiological response.

Recent studies have demonstrated that surgical ventricular septal defect closure in childhood is associated with reduced functional capacity and disruption of the right ventricular force-frequency relationship during exercise. To further describe long-term cardiac function, we performed a non-invasive assessment of cardiac index during exercise in adults having undergone surgery for ventricular septal defect in early childhood. A total of 20 patients (surgical age 2.1±1.4 years, age at examination 22.1±2.2 years) and 20 healthy, matched controls (23.4±2.1 years at examination) underwent continuous supine bicycle ergometry during MRI. Their blood flow was recorded in the ascending aorta and the pulmonary trunk at increasing exercise levels. Cardiac index, retrograde flow, and vessel diameters were determined by blinded, post hoc analyses. The patient group had normal cardiac index at rest (2.9±0.7 L/minute/m2), which was comparable with that of the controls (3.0±0.6 L/minute/m2); however, they had a lower increase in cardiac index during exercise (reaching 7.3±1.3 L/minute/m2 at submaximal exercise) compared with controls (8.2±1.2 L/minute/m2), p<0.05. Patients had a significantly higher ascending aorta retrograde flow than controls at rest and throughout exercise. In the pulmonary artery, the retrograde flow was minimal at rest in both groups, but increased significantly in patients during exercise compared with controls. Young adults with a surgically closed ventricular septal defect have a reduced cardiac index during exercise compared with healthy, young adults. The impaired cardiac index appears to be related to an increasing retrograde flow in the pulmonary artery with progressive exertion.

Age-related Changes In Contractile Function Of Mouse Soleus And Edl Muscles

PURPOSE: Rodent models are often studied to gain insight into aging in human muscle. However, lifespan is not consistent across rodent species or strains, and muscular impairments may not occur at the same relative age (i.e., % survival). We have previously reported ~25% loss of muscle force in rats in early-stage aging, despite smaller (8-15%) declines in muscle mass. It has been suggested that mice, despite shorter lifespans, exhibit less sarcopenia than rats. The goal of the present study was to study the contractile responses of two muscles in aging mice of the same absolute age (24 months), but greater relative age (75% survival, ~ 75 yo in humans), as the rats we have previously studied. METHODS: Ex vivo contractility was studied in the soleus (SOL) and extensor digitorum longus (EDL) muscles of adult (n = 6, 12 mo.) and aged (n = 7, 24 mo.) male, C57Bl/6 mice. All muscles underwent force-frequency relation (FFR) testing and fatigue. Force, muscle quality (force/cross-sectional area (CSA)) and contractile properties for twitch and peak tetanic stimulation were determined for each muscle. RESULTS: Overall analysis of the FFR revealed a significant main effect of muscle and age x frequency and age x frequency x muscle interactions (all P < 0.001). Muscle mass did not differ with age, although there was a trend toward an effect in SOL (P = 0.062). Age significantly (P = 0.035) reduced twitch force in the EDL, but not the SOL, and maximum tetanic force showed no effect of age in either muscle. Twitch contractile properties did not differ with age or muscle. However tetanic rates of force development and relaxation in the EDL were reduced with age (P = 0.065 & 0.098, respectively), differences which became significant when normalized to force (P = 0.018 & 0.035, respectively). No differences in muscle fatigue were found in either muscle. CONCLUSIONS: Relative to aging rats, aging mice exhibit modest changes in muscle mass and force in both the EDL and SOL. Changes in force appear largely determined by changes in mass. Since many studies of human aging indicate that loss of force exceeds loss of strength, these data suggest that mice may not fully recapitulate all aspects of aging human muscle. Future studies should make efforts to examine muscles other than the EDL and SOL to determine if other muscles more closely model aging human muscle.

Vernakalant does not alter early repolarization or contractility in normal and electrically remodelled atria.

Besides the inhibition of the sodium inward current, vernakalant also inhibits the ultra rapid rectifier (IKur) and transient outward current (Ito). Inhibition of these currents increases contractility in canine atrial myocytes and goat atria. We investigated the effect of vernakalant on early repolarization and contractility in normal and electrically remodelled atria. Goats were implanted a pressure catheter, piezoelectric crystals, and electrodes to obtain atrial contractility and effective refractory period (ERP). The active component in pressure distance loops was used to compute the atrial work index (AWI). Experiments were performed in normal and electrically remodelled atria at clinically relevant plasma levels of vernakalant. As a positive control, the Ito/IKur blocker AVE0118 was investigated. Monophasic action potentials were recorded in anaesthetized goats and in explanted hearts to determine changes in action potential morphology. Vernakalant did not affect atrial work loops during sinus rhythm. Likewise vernakalant did not increase atrial fractional shortening or AWI during pacing with fixed heart rate and AV-delay. In contrast, AVE0118 did increase AWI, with a positive force frequency relation. Both in normal and remodelled atria, vernakalant strongly increased ERP but did not prolong early repolarization. In goat atria, vernakalant does not have an atrial positive inotropic effect and does not affect early repolarization. At high rates vernakalant may even have a negative inotropic effect.

The effects of KB-R7943, an inhibitor of reverse Na+/Ca2+ exchange, on the force of contraction of papillary muscles in the heart of the ground squirrel Spermophilus undulatus

We investigated the effect of KB-R7943, an inhibitor of the reverse mode of Na+/Ca2+ exchanger, on the force of isometric contractions, the contractile force–frequency relationship and post-rest potentiation (a qualitative parameter of Ca2+ levels in sarcoplasmic reticulum) in the right ventricle papillary muscles isolated from ground squirrel hearts during summer (June, n = 4) and autumn (October, n = 4) activities. In the presence of 1.8 mM Ca2+at 36°C, 1–1.5 hours-long treatment of the summer papillary muscles with KB-R7943 produced no significant effects on the contractile indices at the majority of stimulation frequencies. In the autumn papillary muscles KB-R7943 induced a 40–50% decrease in the force of contraction (negative inotropic effect) at low stimulation frequencies (0.1–0.3 Hz) without any significant effect at higher stimulation frequencies (0.4–3.0 Hz). Furthermore, in this group, KB-R7943 suppressed the post-rest potentiation of contractility by 50 ± 21% at pause durations exceeding 120 s. These observations indicate that KB-R7943 can affect Ca2+ levels in sarcoplasmic reticulum and that Na+/Ca2+ exchange may contribute to the physiological remodeling of intracellular Ca2+ homeostasis in myocardium of hibernating animals prior their transition to a hypometabolic torpid state.

IGF1 and NRG1 Enhance Proliferation, Metabolic Maturity, and the Force-Frequency Response in hESC-Derived Engineered Cardiac Tissues.

Insulin-like growth factor 1 (IGF1) and neuregulin-1β (NRG1) play important roles during cardiac development both individually and synergistically. In this study, we analyze how 3D cardiac tissue engineered from human embryonic stem cell- (hESC-) derived cardiomyocytes and 2D-plated hESC-cardiomyocytes respond to developmentally relevant growth factors both to stimulate maturity and to characterize the therapeutic potential of IGF1 and NRG1. When administered to engineered cardiac tissues, a significant decrease in active force production of ~65% was measured in all treatment groups, likely due to changes in cellular physiology. Developmentally related processes were identified in engineered tissues as IGF1 increased hESC-cardiomyocyte proliferation 3-fold over untreated controls and NRG1 stimulated oxidative phosphorylation and promoted a positive force-frequency relationship in tissues up to 3 Hz. hESC-cardiomyocyte area increased significantly with NRG1 and IGF1 + NRG1 treatment in 2D culture and gene expression data suggested increased cardiac contractile components in engineered tissues, indicating the need for functional analysis in a 3D platform to accurately characterize engineered cardiac tissue response to biochemical stimulation. This study demonstrates the therapeutic potential of IGF1 for boosting proliferation and NRG1 for promoting metabolic and contractile maturation in engineered human cardiac tissue.

Ant colony optimization for dynamic stability of laminated composite plates

This paper presents the dynamic stability study of laminated composite plates with different force combinations and aspect ratios. Optimum non-diverging stacking is obtained for certain loading combination and aspect ratio. In addition, the stability force is maximized for a definite operating frequency. A dynamic version of the principle of virtual work for laminated composites is used to obtain force-frequency relation. Since dynamic stiffness governs the divergence or flutter, an efficient optimization method is necessary for the response functional and the relevant constraints. In this way, a model based on the ant colony optimization (ACO) algorithm is proposed to search for the proper stacking. The ACO algorithm is used since it treats with large number of dynamic stability parameters. Governing equations are formulated using classic laminate theory (CLT) and von-Karman plate technique. Load-frequency relations are explicitly obtained for fundamental and secondary flutter modes of simply supported composite plate with arbitrary aspect ratio, stacking and boundary load, which are used in optimization process. Obtained results are compared with the finite element method results for validity and accuracy convince. Results revealed that the optimum stacking with stable dynamic response and maximum critical load is in angle-ply mode with almost near-unidirectional fiber orientations for fundamental flutter mode. In addition, short plates behave better than long plates in combined axial-shear load case regarding stable oscillation. The interaction of uniaxial and shear forces intensifies the instability in long plates than short ones which needs low-angle layup orientations to provide required dynamic stiffness. However, a combination of angle-ply and cross-ply stacking with a near-square aspect ratio is appropriate for the composite plate regarding secondary flutter mode.

Fast-to-Slow Transition of Skeletal Muscle Contractile Function and Corresponding Changes in Myosin Heavy and Light Chain Formation in the R6/2 Mouse Model of Huntington's Disease.

Huntington´s disease (HD) is a hereditary neurodegenerative disease resulting from an expanded polyglutamine sequence (poly-Q) in the protein huntingtin (HTT). Various studies report atrophy and metabolic pathology of skeletal muscle in HD and suggest as part of the process a fast-to-slow fiber type transition that may be caused by the pathological changes in central motor control or/and by mutant HTT in the muscle tissue itself. To investigate muscle pathology in HD, we used R6/2 mice, a common animal model for a rapidly progressing variant of the disease expressing exon 1 of the mutant human gene. We investigated alterations in the extensor digitorum longus (EDL), a typical fast-twitch muscle, and the soleus (SOL), a slow-twitch muscle. We focussed on mechanographic measurements of excised muscles using single and repetitive electrical stimulation and on the expression of the various myosin isoforms (heavy and light chains) using dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole muscle and single fiber preparations. In EDL of R6/2, the functional tests showed a left shift of the force-frequency relation and decrease in specific force. Moreover, the estimated relative contribution of the fastest myosin isoform MyHC IIb decreased, whereas the contribution of the slower MyHC IIx isoform increased. An additional change occurred in the alkali MyLC forms showing a decrease in 3f and an increase in 1f level. In SOL, a shift from fast MyHC IIa to the slow isoform I was detectable in male R6/2 mice only, and there was no evidence of isoform interconversion in the MyLC pattern. These alterations point to a partial remodeling of the contractile apparatus of R6/2 mice towards a slower contractile phenotype, predominantly in fast glycolytic fibers.

Hyperthermia and supraspinal fatigue.

Hyperthermia and supraspinal fatigue.

Contractile function and motor unit firing rates of the human hamstrings.

We studied motor unit firing rates (MUFRs) at various voluntary contraction intensities in the hamstrings, one of the only major lower limb muscles to have MUFRs affected by muscle length changes. Within the hamstrings muscle-specific differences have greater impact on MUFRs than length changes, with the biceps femoris having reduced neural drive compared with the semimembranosus-semimembranosus. Comparing our results to other lower limb muscles, flexors have inherently higher firing rate compared with extensors.

Mechanical Stress Conditioning and Electrical Stimulation Promote Contractility and Force Maturation of Induced Pluripotent Stem Cell-Derived Human Cardiac Tissue.

Tissue engineering enables the generation of functional human cardiac tissue with cells derived in vitro in combination with biocompatible materials. Human-induced pluripotent stem cell-derived cardiomyocytes provide a cell source for cardiac tissue engineering; however, their immaturity limits their potential applications. Here we sought to study the effect of mechanical conditioning and electric pacing on the maturation of human-induced pluripotent stem cell-derived cardiac tissues. Cardiomyocytes derived from human-induced pluripotent stem cells were used to generate collagen-based bioengineered human cardiac tissue. Engineered tissue constructs were subjected to different mechanical stress and electric pacing conditions. The engineered human myocardium exhibits Frank-Starling-type force-length relationships. After 2 weeks of static stress conditioning, the engineered myocardium demonstrated increases in contractility (0.63±0.10 mN/mm2 vs 0.055±0.009 mN/mm2 for no stress), tensile stiffness, construct alignment, and cell size. Stress conditioning also increased SERCA2 (Sarco/Endoplasmic Reticulum Calcium ATPase 2) expression, which correlated with a less negative force-frequency relationship. When electric pacing was combined with static stress conditioning, the tissues showed an additional increase in force production (1.34±0.19 mN/mm2), with no change in construct alignment or cell size, suggesting maturation of excitation-contraction coupling. Supporting this notion, we found expression of RYR2 (Ryanodine Receptor 2) and SERCA2 further increased by combined static stress and electric stimulation. These studies demonstrate that electric pacing and mechanical stimulation promote maturation of the structural, mechanical, and force generation properties of human-induced pluripotent stem cell-derived cardiac tissues.

Revealing calcium fluxes by analyzing inhibition dynamics in action potential clamp

In cardiac excitation-contraction coupling (ECC), calcium enters the cytosol via L-type Ca2+ channels (LTCC) and reverse Na+/Ca2+-exchange (NCXrev), or is released from the sarcoplasmic reticulum (SR) by Ca2+-induced Ca2+-release (CICR). The magnitude of Ca2+ influx via the different pathways varies with the state of the cell and is difficult to assess quantitatively, because changes in Ca2+ influx through one pathway affect the others. In rainbow trout ventricular myocytes, the role of the SR has been uncertain for decades. The aim of this work was therefore two-fold: 1) to develop a method to quantify the Ca2+ influx pathways, and 2) to determine the role of CICR from the SR in trout ventricular myocytes. The novelty of our developed method lies in the mathematical analysis of measured transsarcolemmal Ca2+ currents and their impact on the corresponding Ca2+ transient during gradual inhibition of the currents in action potential (AP) clamp. We tested the developed method using an excitation-contraction model and showed that the method was able to recover calcium fluxes from noisy synthetic data. We applied the approach to trout ventricular myocytes and quantified the relative contributions of different Ca2+ influx pathways in ECC and determined the kinetics of these fluxes. Under baseline conditions, NCXrev is the main transmembrane Ca2+ influx pathway contributing 29 ± 6% (of the Ca2+ influx), LTCC 18 ± 7%, and CICR 53 ± 10% to overall Ca2+ transient. Thus, NCXrev is an important regulator of contractility and probably plays a role in the negative force-frequency relationship of trout ventricular preparations. These results demonstrate that trout and neonatal mammalian cardiomyocytes resemble each other not only in terms of morphology and energetics but ECC as well. In summary, the developed method resolves the major problem how to separate highly interconnected fluxes in AP clamp and allows to study Ca2+ fluxes in cardiomyocytes under conditions close to in vivo.

Hemodynamic Characteristics of Patients with Impaired Force Frequency Relationship

Hemodynamic Characteristics of Patients with Impaired Force Frequency Relationship

Molecular Imaging of the Failing Heart: Assessment of Cardiac Sympathetic Nerve and Mitochondrial Function

Patients with heart failure have a high morbidity and mortality despite the advancement of recent heart failure treatment. It is important to evaluate the mechanism of the failing myocardium for decision making appropriate managements or the prediction of prognosis in patients with heart failure. Myocardium mainly utilizes fatty acid or glucose as the energy substrate of oxidative regeneration of ATP in the mitochondria. Intracellular calcium handling, that needs an amount of ATPs in several processes, induces myocardial contraction and relaxation by the sliding of the actin-myosin filament. Moreover, beta adrenal-stimulus also regulates intracellular calcium handling. In the failing myocardium, these components related to the myocardial work are variedly impaired, by various etiologies, including ischemia, inflammation, oxidative stress, metabolic or structural disorder, mechanical stress, or various other factors, and could become the imaging targets. In this review article, we focus on the clinical usefulness of 2 radionuclide imaging in evaluating sympathetic nerve function using myocardial 123I-MIBG SPECT and mitochondrial function using myocardial 99mTc-sestamibi SPECT in the failing heart. We summarize the relationship between each scintigraphic finding derived from the above mentioned tracers and myocardial functional properties of force frequency relations, the molecular mechanism of mitochondrial function, calcium handling, or beta-adrenal signaling in patient with cardiomyopathy.

The Etiology of Muscle Fatigue Differs between Two Electrical Stimulation Protocols.

This study aimed at investigating the mechanisms involved in the force reduction induced by two electrical stimulation (ES) protocols that were designed to activate motor units differently. The triceps surae of 11 healthy subjects (8 men; age, ~28 yr) was activated using ES applied over the tibial nerve. Two ES protocols (conventional [CONV]: 20 Hz, 0.05 ms vs wide-pulse high-frequency [WPHF]: 80 Hz, 1 ms) were performed and involved 40 trains (6 s on-6 s off) delivered at an intensity (IES) evoking 20% of maximal voluntary contraction. To analyze the mechanical properties of the motor units activated at IES, force-frequency relation was evoked before and after each protocol. H-reflex and M-wave responses evoked by the last stimulation pulse were also assessed during each ES protocol. Electromyographic responses (∑EMG) were recorded after each train to analyze the behavior of the motor units activated at IES. Metabolic variables, including relative concentrations of phosphocreatine and inorganic phosphate as well as intracellular pH, were assessed using P-MR spectroscopy during each protocol. Larger H-reflex amplitudes were observed during WPHF as compared with CONV, whereas opposite findings were observed for M-wave amplitudes. Despite this difference, both the force reduction (-26%) and metabolic changes were similar between the two protocols. The CONV protocol induced a rightward shift of the force-frequency relation, whereas a significant reduction of the ∑EMG evoked at IES was observed only for the WPHF. These results suggest that a decreased number of active motor units mainly contributed to WPHF-induced force decrease, whereas intracellular processes were most likely involved in the force reduction occurring during CONV stimulation.

The calcium-frequency response in the rat ventricular myocyte: an experimental and modelling study.

In the majority of species, including humans, increased heart rate increases cardiac contractility. This change is known as the force-frequency response (FFR). The majority of mammals have a positive force-frequency relationship (FFR). In rat the FFR is controversial. We derive a species- and temperature-specific data-driven model of the rat ventricular myocyte. As a measure of the FFR, we test the effects of changes in frequency and extracellular calcium on the calcium-frequency response (CFR) in our model and three altered models. The results show a biphasic peak calcium-frequency response, due to biphasic behaviour of the ryanodine receptor and the combined effect of the rapid calmodulin buffer and the frequency-dependent increase in diastolic calcium. Alterations to the model reveal that inclusion of Ca(2+) /calmodulin-dependent protein kinase II (CAMKII)-mediated L-type channel and transient outward K(+) current activity enhances the positive magnitude calcium-frequency response, and the absence of CAMKII-mediated increase in activity of the sarco/endoplasmic reticulum Ca(2+) -ATPase induces a negative magnitude calcium-frequency response. An increase in heart rate affects the strength of cardiac contraction by altering the Ca(2+) transient as a response to physiological demands. This is described by the force-frequency response (FFR), a change in developed force with pacing frequency. The majority of mammals, including humans, have a positive FFR, and cardiac contraction strength increases with heart rate. However, the rat and mouse are exceptions, with the majority of studies reporting a negative FFR, while others report either a biphasic or a positive FFR. Understanding the differences in the FFR between humans and rats is fundamental to interpreting rat-based experimental findings in the context of human physiology. We have developed a novel model of rat ventricular electrophysiology and calcium dynamics, derived predominantly from experimental data recorded under physiological conditions. As a measure of FFR, we tested the effects of changes in stimulation frequency and extracellular calcium concentration on the simulated Ca(2+) transient characteristics and showed a biphasic peak calcium-frequency relationship, consistent with recent observations of a shift from negative to positive FFR when approaching the rat physiological frequency range. We tested the hypotheses that (1) inhibition of Ca(2+) /calmodulin-dependent protein kinase II (CAMKII)-mediated increase in sarco/endoplasmic reticulum Ca(2+) -ATPase (SERCA) activity, (2) CAMKII modulation of SERCA, L-type channel and transient outward K(+) current activity and (3) Na(+) /K(+) pump dynamics play a significant role in the rat FFR. The results reveal a major role for CAMKII modulation of SERCA in the peak Ca(2+) -frequency response, driven most significantly by the cytosolic calcium buffering system and changes in diastolic Ca(2+) .

The Etiology Of Muscle Fatigue Differs Between Two Electrical Stimulation Protocols

PURPOSE: This study aimed at analyzing from combined electrophysiological and metabolic measurements, the mechanisms involved in force decrease throughout two electrical stimulation (ES) protocols for which the motor units (MUs) recruitment by efferent and/or afferent pathways differs. METHODS: The triceps surae of eleven healthy subjects was activated by ES applied over the tibial nerve. Two ES protocols (CONV: 20 Hz - 0.05 ms vs. WPHF: 80 Hz - 1 ms) were used consisting of 40 trains (6 s on- 6 s off) delivered at an intensity (IES) evoking 20% of maximal voluntary contraction. To assess the contractile properties of the solicited MUs, force-frequency relation was evoked at IES before and after each protocol. For both protocols, a single twitch was delivered at IES after each train, and the force and the electrophysiological responses (∑EMG) were recorded to analyze the behavior of the MUs. Phosphocreatine (PCr) depletion was assessed using 31P-MR spectroscopy. RESULTS: MUs were predominantly recruited through efferent pathways for CONV protocol, whereas afferent pathways were more involved in the MUs recruitment during WPHF. Despite this different MU recruitment, both the force reduction (≈-26%) and PCr depletion (≈8%) were similar between the two protocols. The CONV protocol induced a rightward shift of the force-frequency relationship, whereas a significant reduction of the ∑EMG evoked at IES was observed only for the WPHF. CONCLUSIONS: These results suggest that a decreased number of active MUs mainly contributed to WPHF-induced force decrease while intracellular processes were most likely involved in force decrease during CONV stimulation.