Force potentiation in the MDX mouse

Large forces are the primary mechanism of injury in muscular dystrophy, and muscular dystrophy is especially damaging to type IIB muscle fibers. It was hypothesized that post-tetanic potentiation (PTP) would be down-regulated to prevent damage in Xlinked muscular dystrophy (mdx) mice since PTP increases force and PTP effects are greatest in IIB fibers. PTP experiments were performed on the extensor digitorum longus (EDL) of 50 day old mdx (YM) and C57BL/10 (YC) mice and 10 month old mdx (OM) and C57B1710 (OC) mice. Twitch and tetanic forces were lower in mdx than controls and lower in younger than older mice. Contrary to the hypothesis, PTP was higher in both mdx groups compared to controls. OM potentiated more than any other condition (OM: 29.8%, OC: 23.2%, YM: 21.9%, YC: 17.2%). In accordance with literature PTP increased in the older groups. To explain PTP changes, fiber typing and Western blots for myosin light chain kinase (MLCK) were performed. YM and YC had similar fiber type profiles (2% I, 58% IIX/D and 40% IIB). In accordance with literature but contrary to expected conditions for elevated PTP, OM had a slower fiber type profile (1.7% I, 69% IIX/D and 29% IIB) than OC (0.4% I, 61% IIX/D and 38% IIB). No differences were found in MLCK expression. It seems that PTP is up-regulated to maintain muscle function rather than being down-regulated to prevent muscle damage. Ca""^ transient and myosin phosphorylation measurements would be beneficial in explaining increased PTP seen in this study.

Myocardial Remodeling after Large Infarcts in Rat Converts Post Rest-Potentiation in Force Decay

Background: Post-rest contraction (PRC) of cardiac muscle provides indirect information about the intracellular calcium handling. Objective: Our aim was to study the behavior of PRC, and its underlying mechanisms, in rats with myocardial infarction. Methods: Six weeks after coronary occlusion, the contractility of papillary muscles (PM) obtained from sham-operated (C, n = 17), moderate infarcted (MMI, n = 10) and large infarcted (LMI, n = 14) rats was evaluated, following rest intervals of 10 to 60 seconds before and after incubation with lithium chloride (Li+) substituting sodium chloride or ryanodine (Ry). Protein expression of SR Ca(2+)-ATPase (SERCA2), Na+/Ca2+ exchanger (NCX), phospholamban (PLB) and phospho-Ser(16)-PLB were analyzed by Western blotting. Results: MMI exhibited reduced PRC potentiation when compared to C. Opposing the normal potentiation for C, post-rest decays of force were observed in LMI muscles. In addition, Ry blocked PRC decay or potentiation observed in LMI and C; Li+ inhibited NCX and converted PRC decay to potentiation in LMI. Although MMI and LMI presented decreased SERCA2 (72 +/- 7% and 47 +/- 9% of Control, respectively) and phospho-Ser(16)-PLB (75 +/- 5% and 46 +/- 11%, respectively) protein expression, overexpression of NCX (175 +/- 20%) was only observed in LMI muscles. Conclusion: Our results showed, for the first time ever, that myocardial remodeling after MI in rats may change the regular potentiation to post-rest decay by affecting myocyte Ca(2+) handling proteins. (Arq Bras Cardiol 2012;98(3):243-251)

Considerations for determining the time course of post-activation potentiation

We sought to determine the efficacy of using a continuous time course trial to assess the temporal profile of post-activation potentiation and to determine the time course of potentiation of discrete jump squat kinetic and kinematic variables. Eight physically trained men performed jump squats before and 4, 8, and 12 min after a 5-repetition maximum back squat. Time intervals were assessed in 3 discontinuous trials (each time interval assessed in a separate trial) and in 1 continuous trial (all time intervals assessed in a single trial). Percentage differences between continuous and discontinuous trials at each time interval were mostly insubstantial. Discrete variables displayed a diverse time course (effect size: trivial to large); time to maximal values ranged between 5.00 ± 2.53 min (concentric peak force) and 9.50 ± 2.98 min (eccentric mean force). Eccentric variables (8.58 ± 3.56 min) took longer to peak than concentric variables (6.64 ± 2.93 min) (effect size: small). Individual subjects attained maximal values for kinetic and kinematic variables at different time intervals, yet the 4-min interval typically displayed the greatest magnitude and frequency of potentiation. We conclude that a continuous time course trial does not substantially influence performance of subsequent jumps and is appropriate for determining the temporal profile of potentiation, which is influenced by discrete jump squat kinetic and kinematic variables and individual differences.

Validation of jump squats as a practical measure of post-activation potentiation

To determine if post-activation potentiation (PAP) can augment sports performance, it is pertinent that researchers be confident that any enhancement in performance is attributable to the PAP phenomenon. However, obtaining mechanistic measures of PAP in the daily training environment of highly trained athletes is impractical. We sought to validate jump squats as a practical measure with ecological validity to sports performance against a mechanistic measure of PAP. We assessed the evoked muscle twitch properties of the knee extensors and jump squat kinetics of 8 physically trained males in response to a 5-repetition-maximum back squat conditioning stimulus (CS). Evoked muscle twitch, followed by 3 jump squats, was assessed before and at 4, 8, and 12 min post CS. Time intervals were assessed on separate occasions using a Latin square design. Linear regression was used to determine the relationship between post-pre changes in kinetic variables and muscle twitch peak force (Ft) and twitch rate of force development (RFDt). Large correlations were observed for both concentric relative and absolute mean power and Ft (r = 0.50 ± 0.30) and RFDt (r = 0.56 ± 0.27 and r = 0.58 ± 0.26). Concentric rate of force development (RFD) showed moderate correlations with Ft (r = 0.45 ± 0.33) and RFDt (r = 0.49 ± 0.32). Small-to-moderate correlations were observed for a number of kinetic variables (r = -0.42-0.43 ± 0.32-0.38). Jump squat concentric mean power and RFD are valid ecological measures of muscle potentiation, capable of detecting changes in athletic performance in response to the PAP phenomenon.

Does high intensity squatting elicit post activation potentiation in the squat?

The phenomenon post activation potentiation (PAP) can be defined as an increase in muscle performance following a heavy contraction of a movement that utilizes the same muscle group (i.e. improved 20m sprint following a high intensity squat). However, research has yet to examine if heavy squats elicit PAP on the squat itself. Further, past studies have not yet analyzed if PAP is elicited at a moderate intensity (i.e. 60% one-repetition maximum-1RM).

The role of retinoic acid in long-term memory formation and synaptic plasticity in the mollusc Lymnaea stagnalis

The active metabolite of vitamin A, retinoic acid (RA), is involved in memory formation and hippocampal plasticity in vertebrates. A similar role for retinoid signaling in learning and memory formation has not previously been examined in an invertebrate species. However, the conservation of retinoid signaling between vertebrates and invertebrates is supported by the presence of retinoid signaling machinery in invertebrates. For example, in the mollusc Lymnaea stagnalis the metabolic enzymes and retinoid receptors have been cloned from the CNS. In this study I demonstrated that impairing retinoid signaling in Lymnaea by either inhibiting RALDH activity or using retinoid receptor antagonists, prevented the formation of long-term memory (LTM). However, learning and intermediate-term memory were not affected. An additional finding was that exposure to constant darkness (due to the light-sensitive nature of RA) itself enhanced memory formation. This memory-promoting effect of darkness was sufficient to overcome the inhibitory effects of RALDH inhibition, but not that of a retinoid receptor antagonist, suggesting that environmental light conditions may influence retinoid signaling. Since RA also influences synaptic plasticity underlying hippocampal-dependent memory formation, I also examined whether RA would act in a trophic manner to influence synapse formation and/or synaptic transmission between invertebrate neurons. However, I found no evidence to support an effect of RA on post-tetanic potentiation of a chemical synapse. Retinoic acid did, however, reduce transmission at electrical synapses in a cell-specific manner. Overall, these studies provide the first evidence for a role of RA in the formation of implicit long-term memories in an invertebrate species and suggest that the role of retinoid signaling in memory formation has an ancient origin.

Plasticité présynaptique et gliale à long-terme en réponse à un changement chronique de l’activité synaptique,à la jonction neuromusculaire d’amphibien

La plasticité synaptique est une importante propriété du système nerveux, impliquée dans l’intégration de l’information. Cette plasticité a généralement été décrite par des changements aux niveaux pré et postsynaptiques. Notamment, l’efficacité présynaptique, soit la probabilité de libération de neurotransmetteurs associée au contenu quantique d’une synapse, peut être augmentée ou diminuée selon l’activité antérieure de la synapse. Malgré cette caractérisation, les mécanismes à l’origine de la détermination de l’efficacité présynaptique demeurent obscurs. Également, la plasticité synaptique reste encore mal définie au niveau glial, limitant, de ce fait, notre compréhension de l’intégration de l’information. Pourtant, la dernière décennie a mené à une redéfinition du rôle des cellules gliales. Autrefois reléguées à un rôle de support passif aux neurones, elles sont désormais reconnues comme étant impliquées dans la régulation de la neurotransmission. Notamment, à la jonction neuromusculaire (JNM), les cellules de Schwann périsynaptiques (CSPs) sont reconnues pour moduler l’efficacité présynaptique et les phénomènes de plasticité. Un tel rôle actif dans la modulation de la neurotransmission implique cependant que les CSPs soient en mesure de s’adapter aux besoins changeants des JNMs auxquelles elles sont associées. La plasticité synaptique devrait donc sous-tendre une forme de plasticité gliale. Nous savons, en effet, que la JNM est capable de modifications tant morphologiques que physiologiques en réponse à des altérations de l'activité synaptique. Par exemple, la stimulation chronique des terminaisons nerveuses entraîne une diminution persistante de l’efficacité présynaptique et une augmentation de la résistance à la dépression. À l’opposé, le blocage chronique des récepteurs nicotiniques entraîne une augmentation prolongée de l’efficacité présynaptique. Aussi, compte tenu que les CSPs détectent et répondent à la neurotransmission et qu’elles réagissent à certains stimuli environnementaux par des changements morphologiques, physiologiques et d’expression génique, nous proposons que le changement d'efficacité présynaptique imposé à la synapse, soit par une stimulation nerveuse chronique ou par blocage chronique des récepteurs nicotiniques, résulte en une adaptation des propriétés des CSPs. Cette thèse propose donc d’étudier, en parallèle, la plasticité présynaptique et gliale à long-terme, en réponse à un changement chronique de l’activité synaptique, à la JNM d’amphibien. Nos résultats démontrent les adaptations présynaptiques de l’efficacité présynaptique, des phénomènes de plasticité à court-terme, du contenu mitochondrial et de la signalisation calcique. De même, ils révèlent différentes adaptations gliales, notamment au niveau de la sensibilité des CSPs aux neurotransmetteurs et des propriétés de leur réponse calcique. Les adaptations présynaptiques et gliales sont discutées, en parallèle, en termes de mécanismes et de fonctions possibles dans la régulation de la neurotransmission. Nos travaux confirment donc la coïncidence de la plasticité présynaptique et gliale et, en ce sens, soulèvent l’importance des adaptations gliales pour le maintien de la fonction synaptique.

Short-Term Plasticity in a Computational Model of the Tail-Withdrawal Circuit in Aplysia.

The tail-withdrawal circuit of Aplysia provides a useful model system for investigating synaptic dynamics. Sensory neurons within the circuit manifest several forms of synaptic plasticity. Here, we developed a model of the circuit and investigated the ways in which depression (DEP) and potentiation (POT) contributed to information processing. DEP limited the amount of motor neuron activity that could be elicited by the monosynaptic pathway alone. POT within the monosynaptic pathway did not compensate for DEP. There was, however, a synergistic interaction between POT and the polysynaptic pathway. This synergism extended the dynamic range of the network, and the interplay between DEP and POT made the circuit responded preferentially to long-duration, low-frequency inputs.

THE SITE AND MECHANISM OF ACTION OF BENZODIAZEPINE IN RABBIT HIPPOCAMPUS

The purpose of this study was to determine the effects of benzodiazepine in area CA1 of the hippocampus, and to explore possible mechanisms of action for these agents in this brain area. Two distinctly different benzodiazepine-induced changes in hippocampal physiology have been identified. First, benzodiazepine depresses the population spike recorded in stratum pyramidale, indicating a decrease in action potential generation. Second, benzodiazepine decreases the magnitude of post-tetanic potentiation of the population EPSP recorded in stratum radiatum, and shortens the duration. The effect of benzodiazepine on pyramidal cell excitation was reversed by the GABA antagonis bicuculline, and mimicked by GABA itself. Thus the available evidence is consistent with the hypothesis that benzodiazepine acts by enhancing the effect of GABA in this area. In stratum radiatum, on the other hand, the effect of benzodiazepine on post tetanic potentiation of the population EPSP was not altered by bicuculline although bicuculline did antagonize GABA in this area. In addition, application of GABA, while it caused profound changes in the population EPSP,p, did not cause the same changes that were induced by benzodiazepine. Thus the evidence does not support the hypothesis that benzodiazepine is acting in stratum radiatum by enhancing the effects of GABA. ^

Force potentiation as a modulator of contractile performance: Implications for control of skeletal muscle force and energetics of work

This thesis investigated the modulation of dynamic contractile function and energetics of work by posttetanic potentiation (PTP). Mechanical experiments were conducted in vitro using software-controlled protocols to stimulate/determine contractile function during ramp shortening, and muscles were frozen during parallel incubations for biochemical analysis. The central feature of this research was the comparison of fast hindlimb muscles from wildtype and skeletal myosin light chain kinase knockout (skMLCK-/-) mice that does not express the primary mechanism for PTP: myosin regulatory light chain (RLC) phosphorylation. In contrast to smooth/cardiac muscles where RLC phosphorylation is indispensable, its precise physiological role in skeletal muscle is unclear. It was initially determined that tetanic potentiation was shortening speed dependent, and this sensitivity of the PTP mechanism to muscle shortening extended the stimulation frequency domain over which PTP was manifest. Thus, the physiological utility of RLC phosphorylation to augment contractile function in vivo may be more extensive than previously considered. Subsequent experiments studied the contraction-type dependence for PTP and demonstrated that the enhancement of contractile function was dependent on force level. Surprisingly, in the absence of RLC phosphorylation, skMLCK-/- muscles exhibited significant concentric PTP; consequently, up to ~50% of the dynamic PTP response in wildtype muscle may be attributed to an alternate mechanism. When the interaction of PTP and the catchlike property (CLP) was examined, we determined that unlike the acute augmentation of peak force by the CLP, RLC phosphorylation produced a longer-lasting enhancement of force and work in the potentiated state. Nevertheless, despite the apparent interference between these mechanisms, both offer physiological utility and may be complementary in achieving optimal contractile function in vivo. Finally, when the energetic implications of PTP were explored, we determined that during a brief period of repetitive concentric activation, total work performed was ~60% greater in wildtype vs. skMLCK-/- muscles but there was no genotype difference in High-Energy Phosphate Consumption or Economy (i.e. HEPC: work). In summary, this thesis provides novel insight into the modulatory effects of PTP and RLC phosphorylation, and through the observation of alternative mechanisms for PTP we further develop our understanding of the history-dependence of fast skeletal muscle function.

Duration-dependent effects of the BDNF Val66Met polymorphism on anodal tDCS induced motor cortex plasticity in older adults: a group and individual perspective

The brain derived neurotrophic factor (BDNF) Val66Met polymorphism and stimulation duration are thought to play an important role in modulating motor cortex plasticity induced by non-invasive brain stimulation (NBS). In the present study we sought to determine whether these factors interact or exert independent effects in older adults. Fifty-four healthy older adults (mean age = 66.85 years) underwent two counterbalanced sessions of 1.5 mA anodal transcranial direct current stimulation (atDCS), applied over left M1 for either 10 or 20 min. Single pulse transcranial magnetic stimulation (TMS) was used to assess corticospinal excitability (CSE) before and every 5 min for 30 min following atDCS. On a group level, there was an interaction between stimulation duration and BDNF genotype, with Met carriers (n = 13) showing greater post-intervention potentiation of CSE compared to Val66Val homozygotes homozygotes (n = 37) following 20 min (p = 0.002) but not 10 min (p = 0.219) of stimulation. Moreover, Met carriers, but not Val/Val homozygotes, exhibited larger responses to TMS (p = 0.046) after 20 min atDCS, than following 10 min atDCS. On an individual level, two-step cluster analysis revealed a considerable degree of inter-individual variability, with under half of the total sample (42%) showing the expected potentiation of CSE in response to atDCS across both sessions. Intra-individual variability in response to different durations of atDCS was also apparent, with one-third of the total sample (34%) exhibiting LTP-like effects in one session but LTD-like effects in the other session. Both the inter-individual (p = 0.027) and intra-individual (p = 0.04) variability was associated with BDNF genotype. In older adults, the BDNF Val66Met polymorphism along with stimulation duration appears to play a role in modulating tDCS-induced motor cortex plasticity. The results may have implications for the design of NBS protocols for healthy and diseased aged populations.

Mdx myotubes have normal excitability but show reduced contraction-relaxation dynamics.

The pathogenesis of Duchenne muscular dystrophy (DMD), characterised by lack of the cytoskeletal protein dystrophin, is not completely understood. An early event in the degenerative process of DMD muscle could be a rise in cytosolic calcium concentration. In order to investigate whether this leads to alterations of contractile behaviour, we studied the excitability and contractile properties of cultured myotubes from control (C57BL/10) and mdx mice, an animal model for DMD. The myotubes were stimulated electrically and their motion was recorded photometrically. No significant differences were found between control and mdx myotubes with respect to the following parameters: chronaxy and rheobase (0.33 +/- 0.03 ms and 23 +/- 4 V vs. 0.39 +/- 0.07 ms and 22 +/- 2 V for C57 and mdx myotubes, respectively), tetanisation frequency (a similar distribution pattern was found between 5 and 30 Hz), fatigue during tetanus (found in 35% of both types of myotubes) and post-tetanic contracture. In contrast, contraction and relaxation times were longer (P < 0.005) in mdx (36 +/- 2 and 142 +/- 13 ms, respectively) than in control myotubes (26 +/- 1 and 85 +/- 9 ms, respectively). Together with our earlier findings, these results suggest a decreased capacity for calcium removal in mdx cells leading, in particular, to alterations of muscle relaxation.

Decreased motor unit firing rate in the potentiated tibialis anterior in humans

With repeated activity, force production, rate of force production, and relaxation time are impaired. These are characteristics ofa fatigued muscle (Vandenboom, 2004). However, brief bouts of near maximal to maximal activity results in the increased ability of the muscle to generate force, termed post activation potentiation (P AP)(V andervoort et aI., 1983). The purpose of the present study was to characterize motor unit firing rate (MUFR) in the unfatigued, potentiated tibialis anterior (TA). Using a quadrifilar needle electrode, MUFR was measured during a 5s 50% MVC in which the TA was either potentiated or unpotentiated; monopolar electrodes measured surface parameters. A lOs MVC was used to potentiate the muscle. Firing rate decreased significantly from 20.15±2.9Opps to 18.27±2.99pps, while mean power frequency decreased significantly from 60. 13±7.75 Hz to 53.62±8.56 Hz. No change in root mean square (RMS) was observed. Therefore, in the present study, MUFR decreases in response to a potentiated TA.

Effet d’une pré-sollicitation maximale isométrique des muscles stabilisateurs sur la coordination intermusculaire lors d’un exercice pluriarticulaire épuisant.

Introduction: Les stratégies d’optimisation de la performance chez les athlètes sont de plus en plus exploitées par les entraîneurs et préparateurs physiques. La potentialisation de post- activation (PAP) est reconnue comme étant un phénomène pouvant mener à une augmentation des performances. L’objectif de la présente étude était de donc décrire les effets d’une pré-sollicitation à la hanche sur la coordination inter-musculaire et la performance au cours d’un exercice épuisant. Méthodes: Six athlètes de patins de vitesse de courte piste (3 de sexe masculin et 3 de sexe féminin; âge: 20.2 ± 2.8 ans; moyenne±écart-type) ont exécuté aléatoirement un exercice qui consistait en 2 séries de 9 blocs de squats sautés maximaux, entre-coupés d’un squat isométrique d’une durée de 5 secondes sans pré-sollicitation préalable (CON) et avec une tâche de pré-sollicitation unilatérale de squat isométrique (EXP) contre une barre fixe de 2x3 secondes. Le pic de puissance moyen, l’amplitude et la fréquence moyenne d’EMG, et la vitesse et l’accélération angulaires des premiers et derniers blocs étaient enregistrés. Résultats: La pré-sollicitation isométrique maximale des membres inférieurs n’a pas amélioré de manière significative la performance de sauts et la coordination des muscles stabilisateurs à la hanche. La fréquence spectrale moyenne a néanmoins témoigné de l’implication de stratégies compensatoires du membre inférieur gauche en réponse à la fatigue. Conclusion: La pré-sollicitation des stabilisateurs à la hanche n’augmenterait pas la performance de squats répétés. Par contre, la fréquence moyenne du grand fessier et du tibial antérieur gauche ont suggéré meilleure résistance à la fatigue des muscles du membre inférieur non-dominant avec une pré-sollicitation. Les résultats de la présente étude indiquent donc la pertinence de considérer la pré-sollicitation dans un objectif de performance et de réadaptation sachant que l’asymétrie est omniprésente chez les athlètes et qu’elle est impliquée dans le taux élevé de blessures enregistré chez cette population.