Changes in the time course of potentiated and fatigued contractions of fast motor units in rat muscle

The contraction and relaxation times of the twitches and the last contractions within 32 unfused tetani of FF and 27 unfused tetani of FR motor units in the rat medial gastrocnemius muscle were studied during prolonged activity. The pattern of the MU stimulation included single pulses (to evoke twitches) and series of three trains of stimuli at 40, 50 and 60 Hz (to evoke unfused tetani), repeated 30 times. The analysis concerned changes of force and time parameters at the beginning of activity, during the potentiation and then during the fatigue. It was found that changes of force during the potentiation and the fatigue were mainly accompanied by changes in the course of relaxation. The significant prolongation of the half-relaxation time during the potentiation of either twitches or unfused tetani was revealed in both types of fast MU. The twitch contraction time did not change markedly, whereas significantly shortened in the last contractions of unfused tetani during the potentiation. These changes of time parameters correlated to the increase of the fusion degree. During the fatigue, the time parameters shortened, however, changes of the half-relaxation times were remarkably higher. The shortening of relaxation was responsible for the decrease of the fusion degree. Changes of the fusion index exceeding 0.75 during the potentiation or decreasing below this value during the fatigue, were accompanied by respective appearance or disappearance of the biphasic relaxation.     

Modeling of summation of individual twitches into unfused tetanus for various types of rat motor units.

Repeated stimulation of motor units (MUs) causes an increase of the force output that cannot be explained by linear summation of equal twitches evoked by the same stimulation pattern. To explain this phenomenon, an algorithm for reconstructing the individual twitches, that summate into an unfused tetanus is described in the paper. The algorithm is based on an analytical function for the twitch course modeling. The input parameters of this twitch model are lead time, contraction and half-relaxation times and maximal force. The measured individual twitches and unfused tetani at 10, 20, 30 and 40 Hz stimulation frequency of three rat motor units (slow, fast resistant to fatigue and fast fatigable) are processed. It is concluded that: (1) the analytical function describes precisely the course of individual twitches; (2) the summation of equal twitches does not follow the results from the experimentally measured unfused tetani, the differences depend on the type of the MU and are bigger for higher values of stimulation frequency and fusion index; (3) the reconstruction of individual twitches from experimental tetanic records can be successful if the tetanus is feebly fused (fusion index up to 0.7); (4) both the maximal forces and time parameters of individual twitches subtracted from unfused tetani change and influence the course of each tetanus. A discrepancy with respect to the relaxation phase was observed between experimental results and model prediction for tetani with fusion index exceeding 0.7. This phase was predicted longer than the experimental one for better fused tetani. Therefore, a separate series of physiological experiments and then, more complex model are necessary for explanation of this distinction.     

Model-generated decomposition of unfused tetani of motor units evoked by random stimulation

Unfused tetani of motor units (MUs) evoked by stimulation at variable interpulse intervals at mean frequencies of 20, 25, 33, 40 and 50 Hz were studied using ten functionally isolated fast-type MUs from the medial gastrocnemius muscle of adult Wistar rats. A previously proposed algorithm and computer program for mathematical decomposition of unfused tetani into a series of twitches, representing responses to individual pulses, were used. Analysis of the parameters of the decomposed twitches showed considerable variability in force of successive contractions. These twitches were extremely variable with up to 2-fold higher forces and longer contraction times than a single twitch evoked by one stimulus. However, when the stimulation frequency was decreased, the decomposed twitches became similar to the single twitch with respect to amplitude and contraction time. It was found that the basic contractile parameters of decomposed twitches could be predicted with high accuracy on the basis of the tetanus force level at which the next contraction begins. This analysis of the parameters of decomposed twitches demonstrated that the contractile responses of the muscle fibers to successive action potentials generated by motoneurons are highly variable and depend on the previous MU state.     

Summation of motor unit forces in rat medial gastrocnemius muscle

The summation of contractile forces of motor units (MUs) was analyzed by comparing the recorded force during parallel stimulation of two and four individual MUs or four groups of MUs to the algebraic sum of their individual forces. Contractions of functionally-isolated single MUs of the medial gastrocnemius muscle were evoked by electrical stimulation of thin filaments of the split L5 or L4 ventral roots of spinal nerves. Additionally, contractions of large groups of MUs were evoked by stimuli delivered to four parts of the divided L5 ventral root. Single twitches, 40 Hz unfused tetani, and 150 Hz fused maximum tetani were recorded. In these experimental situations the summation was more effective for unfused tetani than for twitches or maximum tetani. The results obtained for pairs of MUs were highly variable (more- or less-than-linear summation), but coactivation of more units led to progressively weaker effects of summation, which were usually less-than-linear in comparison to the algebraic sums of the individual forces. The variability of the results highlights the importance of the structure of the muscle and the architecture of its MUs. Moreover, the simultaneous activity of fast and slow MUs was considerably more effective than that of two fast units.     

Isometric Muscle Contraction and the Active State: An Analog Computer

From Sandow's excitation-contraction coupling hypothesis and reasonable assumptions I obtain the kinetics of the active state, (AS), and thence, via empirical equations for series elastic and contractile components for frog sartorius around 20 degrees C, the tension, P, and dP/dt vs. time. Assumptions: (a) Rate of Ca(+2) injection is proportional to the Ca gradient, and a permeability, which increases from zero to a limit as the membrane potential rises above a threshold. (b) Released Ca(+2) is bound by the "muscle machinery," M, and removed by a carrier pump. (c) The AS is proportional to the concentration of Ca-M. The kinetic pattern depends mainly upon the mechanism; the time scale was fixed by the amount of Ca(+2) injected. Depending upon the time course and repetition pattern chosen for the action potential, I obtain P and dP/dt, that agree well with experiment, for normal, potentiated, and summed twitches, tetani, and tension redevelopment after a quick release. Upon excitation the AS rises rapidly to 88%, declines thereafter in twitches, but rises slowly in unfused fashion toward 100% in tetani. The knee in dP/dt marks the first maximum in the AS. Potentiators should raise it in tetani as well as in twitches. Velocity and dP/dt show a much higher fusion frequency than P. The model integrates diverse observations. It may be tested by measuring tension and intramyofibrillar Ca(+2) under controlled depolarization.     

Changes in the properties of mechanomyographic signals and in the tension during the fatigue test of rat medial gastrocnemius muscle motor units

The influence of activity-related changes in tension on properties of the mechanomyogram (MMG) was investigated in fast fatigable, fast resistant and slow motor units (MUs). A standard fatigue test was used in which rhythmically repeated unfused tetani were evoked. The amplitudes of the rise in tension of the first and the last contraction within the unfused tetanus and the amplitudes of accompanying signals in MMG were calculated. For fast fatigable MUs a parallel decrease in the amplitudes of both analysed contractions and in the amplitudes of accompanying MMG signals during the fatigue test was observed. For majority of fast resistant MUs at the beginning of the fatigue test a potentiation occurred and this phenomenon increased the tension of the first contraction and of the peak tetanic tension. However, the potentiation coincided also with a decrease of the amplitude of the last contraction in the tension recording of an unfused tetanus. The MMG reflected both, the increase of amplitude of the first contraction and the decrease of the amplitude of the further contractions within the tetanus. The single twitch contraction evoked immediately before and after the fatigue test was additionally recorded. A decrease (fatigue) or an increase (potentiation) of the twitch tension after the fatigue test was reflected by a decrease or an increase in the amplitude of MMG, respectively. However, the fatigue failed to change significantly the time parameters of MMG. To conclude, fatigue and potentiation can occur during activity of fast MUs and both these phenomena involve changes in the amplitude of oscillations in tension of unfused tetani which are reflected in MMG.     

Experimentally verified mathematical approach for the prediction of force developed by motor units at variable frequency stimulation patterns

During normal daily activity, muscle motor units (MUs) develop unfused tetanic contractions evoked by trains of motoneuronal firings at variable interpulse intervals (IPIs). The mechanical responses of a MU to successive impulses are not identical. The aim of this study was to develop a mathematical approach for the prediction of each response within the tetanus as well as the tetanic force itself. Experimental unfused tetani of fast and slow rat MUs, evoked by trains of stimuli at variable IPIs, were decomposed into series of twitch-shaped responses to successive stimuli using a previously described algorithm. The relationships between the parameters of the modeled twitches and the tetanic force level at which the next response begins were examined and regression equations were derived. Using these equations, profiles of force for the same and different stimulation patterns were mathematically predicted by summating modeled twitches. For comparison, force predictions were made by the summation of twitches equal to the first one. The recorded and the predicted tetanic forces were compared. The results revealed that it is possible to predict tetanic force with high accuracy by using regression equations. The force predicted in this way was much closer to the experimental record than the force obtained by the summation of equal twitches, especially for slow MUs. These findings are likely to have an impact on the development of realistic muscle models composed of MUs, and will assist our understanding of the significance of the neuronal code in motor control and the role of biophysical processes during MU contractions.     

Initial force production before sag is enhanced by prior contraction followed by a 3-minute rest period in fast motor units of the rat medial gastrocnemius

Unfused tetanic contractions evoked in fast motor units exhibit extra-efficient force production at the onset of contraction, an effect called “boost”. Boost is diminished in subsequent contractions if there is a short rest period between contractions, but can be re-established with a longer period of rest. We tested the hypothesis that contractile activity and rest could enhance boost-related metrics. Two sets of 3 unfused tetani were evoked 3 min apart in fast fatigable (FF) and fast fatigue-resistant (FR) motor units of the rat medial gastrocnemius. The greatest changes occurred in the first unfused tetanic contractions. Relative to the first contraction in the first set, the first contraction in the second set exhibited higher peak force during boost in a subset of motor units (76% of FF and 48% of FR). Enhanced force during boost was influenced by interaction of slowing of twitch contraction time (up to 20% and 25%, for FF and FR motor units, respectively), half-relaxation time (up to 37% and 49% for FF and FR motor units, respectively), and potentiation of the first twitch (up to 13% and 5% for FF and FR motor units, respectively). Examination of twitches evoked between sets suggested opportunity for greater enhancement of boost with shorter intervening rest periods. The phenomenon of enhanced boost following motor unit activity may interest sports scientists.     

Experimentally verified model of mechanomyograms recorded during single motor unit contractions

The aim of the paper is to create a model which enables to observe the mechanomyographic (MMG) wave generated during single motor unit contractions in a muscle, while the muscle is immersed in paraffin oil. The muscle model is described as a rheological membrane. Both the muscle and the medium models have been built by using Stiff-Finite-Element-Method (SFEM), which allows one to simulate the muscle surface displacement and the acoustic propagation of this effect in the oil. Such a modelling enables one to determine the impact of the rheological properties of the liquid environment on the shape of the MMG wave. In order to verify the model, the MMG signals and the contraction forces have been recorded in vivo from the medial gastrocnemius muscle of a rat. In these experiments single motor units were stimulated with various stimulation frequencies. A piezotransducer, immersed in paraffin oil, has been used to record the MMG signal recording. The signals recorded during individual twitches of the motor units have been used to estimate the parameters of the model. Subsequently, the model has been experimentally verified. The signals recorded in experiments during unfused and fused tetani have been compared with the simulated model responses in the analogous stimulation program. It has been observed that the MMG signals obtained with the proposed linear model have been consistent with the results of in vivo experiments.     

Effects of caffeine at different temperatures on contractile properties of slow-twitch and fast-twitch rat muscles

The slow-twitch soleus muscle (SOL) exhibits decreased twitch tension (cold depression) in response to a decreased temperature, whereas the fast-twitch extensor digitorum longus (EDL) muscle shows enhanced twitch tension (cold potentiation). On the other hand, the slow-twitch SOL muscle is more sensitive to twitch potentiation and contractures evoked by caffeine than the fast-twitch EDL muscle. In order to reveal the effects of these counteracting conditions (temperature and caffeine), we have studied the combined effects of temperature changes on the potentiation effects of caffeine in modulating muscle contractions and contractures in both muscles. Isolated muscles, bathed in a Tyrode solution containing 0.1-60 mM caffeine, were stimulated directly and isometric single twitches, fused tetanic contractions and contractures were recorded at 35 degrees C and 20 degrees C. Our results showed that twitches and tetani of both SOL and EDL were potentiated and prolonged in the presence of 0.3-10 mM caffeine. Despite the cold depression, the extent of potentiation of the twitch tension by caffeine in the SOL muscle at 20 degrees C was by 10-15 % higher than that at 35 degrees C, while no significant difference was noted in the EDL muscle between both temperatures. Since the increase of twitch tension was significantly higher than potentiation of tetani in both muscles, the twitch-tetanus ratio was enhanced. Higher concentrations of caffeine induced contractures in both muscles; the contracture threshold was, however, lower in the SOL than in the EDL muscle at both temperatures. Furthermore, the maximal tension was achieved at lower caffeine concentrations in the SOL muscle at both 35 degrees C and 20 degrees C compared to the EDL muscle. These effects of caffeine were rapidly and completely reversed in both muscles when the test solution was replaced by the Tyrode solution. The results have indicated that the potentiation effect of caffeine is both time- and temperature-dependent process that is more pronounced in the slow-twitch SOL than in the fast-twitch EDL muscles.     

Estimation of contractile parameters of successive twitches in unfused tetanic contractions of single motor units – A proof-of-concept study using ultrafast ultrasound imaging in vivo

During a voluntary contraction, motor units (MUs) fire a train of action potentials, causing summation of the twitch forces, resulting in fused or unfused tetanus. Twitches have been important in studying whole-muscle contractile properties and differentiation between MU types. However, there are still knowledge gaps concerning the voluntary force generation mechanisms. Current methods rely on the spike-triggered averaging technique, which cannot track changes in successive twitches’ properties in response to individual neural firings. This study proposes a method that estimates successive twitches contractile parameters of single MUs during low force voluntary isometric contractions in human biceps brachii. We used a previously developed ultrafast ultrasound imaging method to estimate unfused tetanic activity signals of single MUs. A twitch decomposition model was used to decompose unfused tetanic activity signals into individual twitches. This study found that the contractile parameters varied within and across MUs. There was an association between the inter-spike interval and the contraction time (r = 0.49, p < 0.001) and the half-relaxation time (r = 0.58, p < 0.001), respectively. The method shows the proof-of-concept to study MU contractile properties of individual twitches in vivo, which can provide further insights into the force generation mechanisms of voluntary contractions and response to individual neural discharges.     

Summation of slow motor unit forces at constant and variable interpulse intervals in rat soleus muscle

Effects of the summation of forces generated by functionally isolated slow-twitch motor units (MU) of the rat soleus muscle were examined in this study. Initially, the twitch, fused tetanic and unfused tetanic contractions evoked by trains of stimuli at variable interpulse intervals were recorded for each MU. Then, two, three or four MUs were co-activated, and the recorded forces were compared to the algebraic sum of the forces of individual MUs. The mean cumulative force of twitches and the mean cumulative force of fused tetani were not statistically different from the respective algebraic sums of forces, which revealed a high degree of linearity in the summation. However, relaxation of the recorded tetanic contractions (either fused or unfused) was faster than that predicted by the linear summation of individual contractions. Moreover, for twitch and tetanic contractions, a tendency to shorten relaxation with an increasing number of co-active MUs was noted. The results indicate that forces of rat soleus slow MUs sum up more linearly than in the respective cat muscle as well as more linearly than for fast MUs in the medial gastrocnemius muscle.     

Slow changes and wiener analysis of nonlinear summation in contractions of cat muscles

With regular trains of stimuli at a high frequency, the contribution of each stimulus to the force generated over time declines from the second to about the tenth stimulus, but then begins to increase again. This late increase is referred to as tetanic potentiation in analogy with the post-tetanic potentiation of the twitch after such a period of stimulation. With regular trains of stimuli at a low frequency, a progressive decrease in the essentially unfused twitches (negative staircase) is observed in the slow soleus muscle of the cat, while a progressive increase (positive staircase) is observed for the fast plantaris muscle. The time constant for the approximately exponential changes observed is on the order of 10 s. Random trains of stimuli were applied at intermediate frequencies and analyzed in terms of general methods of analysis for nonlinear systems. Systematic decreases in the magnitude and increases in the time course of the average tension per stimulus were observed with increasing mean rates of stimulation. Similar changes were observed for short intervals between stimuli within a given random train at a constant mean rate. These changes can be described in terms of an early depression and a later facilitation described in the previous papers in this series.     

Analysis of the unfused tetanus course in fast motor units of the rat medial gastrocnemius muscle

The course of unfused tetani with the sag effect in fast motor units of rat medial gastrocnemius was studied. The analysis of the course of successive contractions within these tetani showed that the high peak force at the beginning of tetanus before the sag resulted from temporary, very efficient sum mation of contractions at this phase, both in FF (fast fatigable) and FR (fast resistant to fatigue) units. The process of summation developed in spite of parallel shortening of the contraction and relaxation. The peak of tetanus force was visible on the average at the 2nd contraction in FF units and at the 5th contraction in FR units. After the tetanus peak the process of the efficient summation was completed and the force decreased what was visible as a sag. In the following part of the tetanus, mainly in FF units, the potentiation occurred and the force of successive contractions increased. The rise of force was visible in spite of shortening of the contraction time and was due to prolongation of the relaxation in this part of the tetanus. These observations indicated that the processes of the summation of successive contractions before sag and during the potentiation underwent different mechanisms discussed in this paper. Considerable release of Ca2+ ions from the sarcoplasmic reticulum was proposed as a possible mechanism responsible for a very efficient summation at the beginning of the tetanus whereas phosphorylation of regulatory light chain of myosin (RLC) in muscle fibers was considered as the reason of potentiation. Moreover, the present analysis revealed that previously found differences in tetani profiles between FF and FR units resulted from faster development of described changes in the course of contractions summating into the tetanus in FF motor units.     

Estimation of the error between experimental tetanic force curves of MUs of rat medial gastrocnemius muscle and their models by summation of equal successive contractions

More accurate muscle models require appropriate modelling of individual twitches of motor units (MUs) and their unfused tetanic contractions. It was shown in our previous papers, using a few MUs, that modelling of unfused tetanic force curves by summation of equal twitches is not accurate, especially for slow MUs. The aim of this study was to evaluate this inaccuracy using a statistical number of MUs of the rat medial gastrocnemius muscle (15 of slow, 15 of fast resistant and 15 of fast fatigable type). Tetanic contractions were evoked by trains of 41 stimuli at random interpulse intervals and different mean frequencies, resembling discharge patterns observed during natural muscle activity. The tetanic curves were calculated by the summation of equal twitches according to the respective experimental patterns. The previously described 6-parameter analytical function for twitch modelling was used. Comparisons between the experimental and the modelled curves were made using two coefficients: the fit coefficient and the area coefficient. The errors between modelled and experimental tetanic forces were substantially different between the three MU types. The error was the most significant for slow MUs, which develop much higher forces in real contractions than could be predicted based on the summation of equal twitches, while the smallest error was observed for FF MUs – their recorded tetanic forces were similar to those predicted by modelling. The obtained results indicate the importance of the inclusion of the type-specific non-linearity in the summation of successive twitch-like contractions of MUs in order to increase the reliability of modelling skeletal muscle force.     

The function of motor nerves innervating slow tonic skeletal muscle in hens with delayed neuropathy induced by tri-o-tolyl phosphate

The reduction of neurogenic posttetanic potentiation in the slow twitch, soleus muscle is an index of impaired motor nerve function in cats with organophosphate-induced neuropathy. We have applied the measurement of posttetanic potentiation to study the functional state of the slow, tonic, plantaris muscle and its motor innervation in adult White Leghorn hens with tri-o-tolyl phosphate (TOTP)-induced neuropathy. At suitable intervals following single oral doses of vehicle or TOTP (500 mg/kg), nerve conduction velocity and posttetanic potentiation were measured in anesthetized hens. Conduction in the sciatic nerve was not altered by TOTP. The plantaris muscle of birds treated with vehicle (peanut oil) either failed to contract or responded to nerve stimulation at 0.4 Hz with very small twitches. Following nerve stimulation at frequencies inducing tetanus (50-140 Hz), the muscles responded with large, slow twitches that gradually decayed in amplitude. The area under the curve formed by the amplitude of these twitches over time (posttetanic potentiation) was directly proportional to the frequency and duration of nerve stimulation. In hens at 1,2, and 4 weeks following treatment with TOTP, the average amount of posttetanic potentiation was reduced concomitantly with the development of ataxia, paralysis, and pathological changes in the peripheral nerves. This difference between vehicle- and TOTP-treated hens was not significant, owing to large interbird variations. Since TOTP-treated hens showed greater disturbances in gait following moderate exercise, the fatigue of posttetanic potentiation with periodic neuronal stimulation was measured.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of temperature on post-tetanic potentiation in human dorsiflexor muscles

The effect of temperature on post-tetanic potentiation (PTP) has been examined in the muscles of small mammals but not in human skeletal muscle. We examined PTP in the ankle dorsiflexor muscles of 10 young men by evoking twitches before and after a 7-second tetanus at 100 Hz in a control (room air approximately 21 degrees C) condition and after immersion of the lower leg in warm (45 degrees C) and cold (10 degrees C) water baths for 30 min. Exposure to cold decreased tetanus and pre-tetanus twitch peak torque, but increased rise time, half-relaxation time, and muscle action potential (M-wave) amplitude; exposure to warm water had little effect. PTP was smallest in cold exposure 5 s post-tetanus, but persisted throughout the 12 min test period, whereas PTP had subsided by 6 min post-tetanus in control and warm exposures. M-wave amplitude initially decreased after exposure to warm water, recovered, then decreased again by 11 min post-tetanus. In contrast, exposure to cold had no initial effect but did increase the M-wave amplitude during the last half of the 12 min test period, similar to that seen in the control. The greatest immediate decrease in rise time and half-relaxation time was observed in the control; however, by 12 min post-tetanus warm exposure showed the greatest increase in rise time and half-relaxation time above pre-tetanus values. The decrease in the unpotentiated twitch torque with cooling in human dorsiflexors is typical for muscles with a predominance of type I (slow) fibres. The effect of cold on PTP is similar to that seen previously in mammalian muscles with a predominance of type II (fast) fibres, although the underlying mechanism of the cooling effect appears to differ.     

Post-tetanic potentiation of miniature end-plate potential frequency at neuromuscular junction of the rat soleus muscle

1. Changes in miniature end-plate potential (m.e.p.p.) frequency by repetitive nerve stimulation were examined in the rat soleus muscle. 2. The increase of m.e.p.p. frequency was induced by repetitive stimulation and persisted for several minutes after the tetanus. That is, post-tetanic potentiation (PTP) of neuromuscular transmission was first demonstrated here in the rat soleus muscle. 3. The time course of the decay of m.e.p.p. frequency after the tetanus showed a double exponential curve which consisted of a fast decaying component (augmentation) and a slow decaying component (potentiation). 4. The magnitude of PTP depended on the stimulation frequency and its duration. It increased with the increase of duration and was at its maximum at a frequency of 100 Hz. 5. No PTP was elicited by repetitive stimulation under conditions in which end-plate potential (e.p.p.) was completely suppressed, and, moreover, m.e.p.p. frequency tended to decrease after the tetanus.     

Isometric contractile properties and instantaneous stiffness of amphibian skeletal muscle in the temperature range from 0 to 20 degrees C

The isometric contractile properties of frog (Rana pipiens) and toad (Bufo bufo) sartorii have been studied over the temperature range from 0 to 20 degrees C. The isometric twitch tension was found to vary considerably between these two species and between muscles in the same species. Between 0 and 4 degrees C there was very little change in maximum isometric twitch tension. Between 4 and 12 degrees C several muscles from frog or toad showed a potentiation of twitch tension whereas others showed a decline. Over this temperature range the toad sartorii consistently demonstrated a greater potentiation. By 12 degrees C a steady decline in twitch tension in both muscles was seen as the temperature range the toad sartorii consistently demonstrated a greater potentiation. By 12 degrees C a steady decline in twitch tension in both muscles was seen as the temperature approached 20 degrees C. The maximum isometric tetanic tension recorded between 18 and 20 degrees C increased fractionally to an average of 1.504 +/- 0.029 (n = 4) for frog sartorii and to 1.377 +/- 0.008 (n = 5) for toad sartorii. The time to peak twitch tension and the half-relaxation time decreased markedly with an increase in temperature. Moreover, the half-relaxation time was reduced by a greater proportion than the time to peak twitch tension. Measurements of instantaneous stiffness by controlled velocity releases from the plateau of isometric tetani revealed that the large increase in isometric tetanus tension as the muscle was warmed was not accompanied by a corresponding increase in the total number of active cross-bridges. The possibility that a decreased availability of intracellular Ca2+ ions at the contractile sites contributing to the fall of isometric twitch tension at elevated temperatures is discussed. The possibility exists that at elevated temperatures a change inthe intrinsic contractile ability of the muscle occurs which produces an increased tension per cross-bridge.     

Phase-Dependent Effects of Stimuli Locked to Oscillatory Activity in Cultured Cortical Networks

The archetypal activity pattern in cultures of dissociated neurons is spontaneous network-wide bursting. Bursts may interfere with controlled activation of synaptic plasticity, but can be suppressed by the application of stimuli at a sufficient rate. We sinusoidally modulated (4 Hz) the pulse rate of random background stimulation (RBS) and found that cultures were more active, burst less frequently, and expressed oscillatory activity. Next, we studied the effect of phase-locked tetani (four pulses, 200 s⁻¹) on network activity. Tetani were applied to one electrode at the peak or trough of mRBS stimulation. We found that when tetani were applied at the peak of modulated RBS (mRBS), a significant potentiation of poststimulus histograms (PSTHs) occurred. Conversely, tetani applied at the trough resulted in a small but insignificant depression of PSTHs. In addition to PSTHs, electrode-specific firing rate profiles within spontaneous bursts before and after mRBS were analyzed. Here, significant changes in firing rate profiles were found only for stimulation at the peak of mRBS. Our study shows that rhythmic activity in culture is possible, and that the network responds differentially to strong stimuli depending on the phase at which they are delivered. This suggests that plasticity mechanisms may be differentially accessible in an oscillatory state.