Cyclic Length Changes Research Articles

The importance of muscle activation on the interpretation of muscle mechanical performance.

The work loop technique was developed to assess muscle performance during cyclical length changes with phasic activation, simulating the in vivo conditions of many muscles, particularly during locomotion. To estimate muscle function in vivo, the standard approach involves subjecting a muscle to length trajectories and activation timings derived from in vivo measurements, whilst simultaneously measuring force. However, the stimulation paradigm typically used, supramaximal, 'square-wave' stimulation, does not accurately reflect the graded intensity of activation observed in vivo. While the importance of the timing and duration of stimulation within the cycle on estimates of muscle performance has long been established, the importance of graded muscle activation has not been investigated. In this study, we investigated how the activation pattern affects muscle performance by comparing square-wave, supramaximal activation with a graded in vivo activation pattern. First, we used in vivo electromyography-derived activation patterns and fibre strains from the rabbit digastric muscle during mastication and replayed them in situ. Second, we used Hill-type musculoskeletal model-derived activation patterns and fibre strains in a trotting mouse, replayed ex vivo in the soleus (SOL) and extensor digitorum longus (EDL) muscles. In the rabbit digastric muscle, square-wave activation led to an 8-fold higher estimate of net power, compared with the in vivo graded activation pattern. Similarly, in the mouse SOL and EDL, supramaximal, square-wave activation resulted in significantly greater positive and negative muscle work. These findings highlight that realistic interpretations of in vivo muscle function rely upon more accurate representations of muscle activation intensity.

The mechanical properties of the mantle muscle of European cuttlefish (Sepia officinalis).

The circular muscles surrounding the mantle cavity of European cuttlefish (Sepia officinalis) generate the mechanical power to compress the cavity, forcing a jet of water out of the funnel, propelling the animal during jet propulsion swimming. During ontogeny, jetting frequency decreases in adults compared with juveniles, and this is expected to be reflected in the contractile properties of the locomotory muscles. To develop greater insight into how the locomotion of these animals is powered during ontogeny, we determined the mechanical properties of bundles of muscle fascicles during isometric, isotonic and cyclic length changes in vitro, at two life stages: juveniles and adults. The twitch kinetics were faster in juveniles than in adults (twitch rise time 257 ms compared with 371 ms; half-twitch relaxation 257 ms compared with 677 ms in juveniles and adults, respectively); however, twitch and tetanic stress, the maximum velocity of shortening and curvature of the force-velocity relationship did not differ. Under cyclic conditions, net power exhibited an inverted U-shaped relationship with cycle frequency in both juveniles and adults; the frequency at which maximum net power was achieved was shifted to lower cycle frequencies with increased maturity, which is consistent with the slower contraction and relaxation kinetics in adults compared with juveniles. The cycle frequency at which peak power was achieved during cyclical contractions in vitro was found to match that seen in vivo in juveniles, suggesting power is being maximised during jet propulsion swimming.

Timing precision in fly flight control: integrating mechanosensory input with muscle physiology.

Animals rapidly collect and act on incoming information to navigate complex environments, making the precise timing of sensory feedback critical in the context of neural circuit function. Moreover, the timing of sensory input determines the biomechanical properties of muscles that undergo cyclic length changes, as during locomotion. Both of these issues come to a head in the case of flying insects, as these animals execute steering manoeuvres at timescales approaching the upper limits of performance for neuromechanical systems. Among insects, flies stand out as especially adept given their ability to execute manoeuvres that require sub-millisecond control of steering muscles. Although vision is critical, here I review the role of rapid, wingbeat-synchronous mechanosensory feedback from the wings and structures unique to flies, the halteres. The visual system and descending interneurons of the brain employ a spike rate coding scheme to relay commands to the wing steering system. By contrast, mechanosensory feedback operates at faster timescales and in the language of motor neurons, i.e. spike timing, allowing wing and haltere input to dynamically structure the output of the wing steering system. Although the halteres have been long known to provide essential input to the wing steering system as gyroscopic sensors, recent evidence suggests that the feedback from these vestigial hindwings is under active control. Thus, flies may accomplish manoeuvres through a conserved hindwing circuit, regulating the firing phase-and thus, the mechanical power output-of the wing steering muscles.

Isometric placement of the augmentation braid is not attained reliably in contemporary ACL suture repair

BackgroundTo assess if during arthroscopic braid-augmented ACL suture repair (ACLSR), the actual positions of the augmentation braids' tunnels corresponded with the positions of their intended and targeted isometric points, and to test the hypothesis that there would be no dispersion in actual positions of the augmentation braids' tunnels compared to their intended and targeted isometric points. MethodsIn 12 human cadaveric knees, the positions of the augmentation braids' tunnels and their intended and targeted isometric points relative to a femoral and tibial grid were analysed. Furthermore, vector length between these positions was calculated to assess the accuracy and precision of the augmentation braids' tunnel placement. ResultsThere was dispersion for all of the augmentation braids' tunnel positions compared to their intended isometric points. The femoral and tibial vector lengths (mean ± SD (range)) were 2.9 ± 1.0 (1.1–4.1) and 7.1 ± 2.0 (3.2–9.8) mm respectively. ConclusionIn augmented ACLSR, with the ruptured ACL in situ, there was dispersion of the positions of the actual small diameter femoral and tibial augmentation braids' tunnels away from their desired isometric points. Clinical relevanceThe extent of dispersion of the position of both the femoral and tibial tunnels away from their intended isometric positions may cause cyclic length changes with knee motion. An ACLSR with static braid augmentation will thus be vulnerable to cyclic stretching-out. The difficulty of obtaining an isometric tunnel combination for the small diameter augmentation braid may influence the clinician's choice between non-, static or dynamic augmented ACLSR techniques.

X-Ray Diffraction Resolves How Actin-Myosin Spacing Explains the Differences of Two Muscles with Identical Steady State Properties

The actin-myosin lattice underlying muscle is dynamic. Radial lattice spacing changes during activation and during periodic axial strain like muscle experiences during locomotion. Actin-myosin spacing in muscle affects crossbridge binding rates and the steady-state length-tension properties of muscle. The interaction of macroscopic strain, lattice spacing and binding kinetics raises the possibility that small changes in lattice spacing could have profound consequences for the dynamic behavior of muscle. To study the functional implications of lattice spacing, we examined two muscles in Blaberus discoidalis. These muscles have nearly identical length-tension and force-velocity relationships and share innervation. However, when periodically activated under identical cyclical length changes, one of them does net positive mechanical work while the other is dissipative. Using the BioCAT x-ray beamline at the Advanced Photon Source at Argonne National Lab, we measured lattice spacing during dynamic oscillations and isometric twitches. We found passive lattice spacing in the two muscles to be 50.01+/−1.54 nm and 50.51+/−.6 nm (95% CI) with the second muscle's spacing consistently higher (p≈10−5). We found that in the isometric twitch case, the first muscle's lattice spacing increased to 51.15+/−1.7 while the other did not change significantly (51.3+/−.65), meaning that the spacing difference vanishes under steady state, activated conditions (p≈.19). A difference in spacing under passive conditions, but not active, means that during cyclic length changes the lattice spacing changes were larger in one muscle. In the muscle with the larger dynamic range, lattice spacing correlates with the timing of increased stress and positive mechanical work. A 1 nm transient difference in actin-myosin spacing mediates the dynamic difference between the two muscles (motor vs. brake) but also predicts their identical quasi-static properties, allowing nanoscale structure to influence macroscopic muscle function.

The effects of elevated levels of sodium bicarbonate (NaHCO3) on the acute power output and time to fatigue of maximally stimulated mouse soleus and EDL muscles

This study examined the effects of elevated buffer capacity [~32 mM HCO₃(-)] through administration of sodium bicarbonate (NaHCO₃) on maximally stimulated isolated mouse soleus (SOL) and extensor digitorum longus (EDL) muscles undergoing cyclical length changes at 37 °C. The elevated buffering capacity was of an equivalent level to that achieved in humans with acute oral supplementation. We evaluated the acute effects of elevated [HCO₃(-)] on (1) maximal acute power output (PO) and (2) time to fatigue to 60 % of maximum control PO (TLIM60), the level of decline in muscle PO observed in humans undertaking similar exercise, using the work loop technique. Acute PO was on average 7.0 ± 4.8 % greater for NaHCO₃-treated EDL muscles (P < 0.001; ES = 2.0) and 3.6 ± 1.8 % greater for NaHCO₃-treated SOL muscles (P < 0.001; ES = 2.3) compared to CON. Increases in PO were likely due to greater force production throughout shortening. The acute effects of NaHCO₃ on EDL were significantly greater (P < 0.001; ES = 0.9) than on SOL. Treatment of EDL (P = 0.22; ES = 0.6) and SOL (P = 0.19; ES = 0.9) with NaHCO₃ did not alter the pattern of fatigue. Although significant differences were not observed in whole group data, the fatigability of muscle performance was variable, suggesting that there might be inter-individual differences in response to NaHCO₃ supplementation. These results present the best indication to date that NaHCO₃ has direct peripheral effects on mammalian skeletal muscle resulting in increased acute power output.

Tendon material properties vary and are interdependent among turkey hindlimb muscles

The material properties of a tendon affect its ability to store and return elastic energy, resist damage, provide mechanical feedback and amplify or attenuate muscle power. While the structural properties of a tendon are known to respond to a variety of stimuli, the extent to which material properties vary among individual muscles remains unclear. We studied the tendons of six different muscles in the hindlimb of Eastern wild turkeys to determine whether there was variation in elastic modulus, ultimate tensile strength and resilience. A hydraulic testing machine was used to measure tendon force during quasi-static lengthening, and a stress-strain curve was constructed. There was substantial variation in tendon material properties among different muscles. Average elastic modulus differed significantly between some tendons, and values for the six different tendons varied nearly twofold, from 829±140 to 1479±106 MPa. Tendons were stretched to failure, and the stress at failure, or ultimate tensile stress, was taken as a lower-limit estimate of tendon strength. Breaking tests for four of the tendons revealed significant variation in ultimate tensile stress, ranging from 66.83±14.34 to 112.37±9.39 MPa. Resilience, or the fraction of energy returned in cyclic length changes was generally high, and one of the four tendons tested was significantly different in resilience from the other tendons (range: 90.65±0.83 to 94.02±0.71%). An analysis of correlation between material properties revealed a positive relationship between ultimate tensile strength and elastic modulus (r(2)=0.79). Specifically, stiffer tendons were stronger, and we suggest that this correlation results from a constrained value of breaking strain, which did not vary significantly among tendons. This finding suggests an interdependence of material properties that may have a structural basis and may explain some adaptive responses observed in studies of tendon plasticity.

Automatic tracking of medial gastrocnemius fascicle length during human locomotion

During human locomotion lower extremity muscle-tendon units undergo cyclic length changes that were previously assumed to be representative of muscle fascicle length changes. Measurements in cats and humans have since revealed that muscle fascicle length changes can be uncoupled from those of the muscle-tendon unit. Ultrasonography is frequently used to estimate fascicle length changes during human locomotion. Fascicle length analysis requires time consuming manual methods that are prone to human error and experimenter bias. To bypass these limitations, we have developed an automatic fascicle tracking method based on the Lucas-Kanade optical flow algorithm with an affine optic flow extension. The aims of this study were to compare gastrocnemius fascicle length changes during locomotion using the automated and manual approaches and to determine the repeatability of the automated approach. Ultrasound was used to examine gastrocnemius fascicle lengths in eight participants walking at 4, 5, 6, and 7 km/h and jogging at 7 km/h on a treadmill. Ground reaction forces and three dimensional kinematics were recorded simultaneously. The level of agreement between methods and the repeatability of the automated method were quantified using the coefficient of multiple correlation (CMC). Regardless of speed, the level of agreement between methods was high, with overall CMC values of 0.90 ± 0.09 (95% CI: 0.86-0.95). Repeatability of the algorithm was also high, with an overall CMC of 0.88 ± 0.08 (95% CI: 0.79-0.96). The automated fascicle tracking method presented here is a robust, reliable, and time-efficient alternative to the manual analysis of muscle fascicle length during gait.

A spatially explicit model of muscle contraction explains a relationship between activation phase, power and ATP utilization in insect flight

Using spatially explicit, stochastically kinetic, molecular models of muscle force generation, we examined the relationship between mechanical power output and energy utilization under differing patterns of length change and activation. A simulated work loop method was used to understand prior observations of sub-maximal power output in the dominant flight musculature of the hawkmoth Manduca sexta L. Here we show that mechanical work output and energy consumption (via ATP) vary with the phase of activation, although they do so with different phase sensitivities. The phase relationship for contraction efficiency (the ratio of power output to power input) differs from the phase relationships of energy consumption and power output. To our knowledge, this is the first report to suggest that ATP utilization by myosin cross-bridges varies strongly with the phase of activation in muscle undergoing cyclic length changes.

A modified Hill muscle model that predicts muscle power output and efficiency during sinusoidal length changes

The power output of a muscle and its efficiency vary widely under different activation conditions. This is partially due to the complex interaction between the contractile component of a muscle and the serial elasticity. We investigated the relationship between power output and efficiency of muscle by developing a model to predict the power output and efficiency of muscles under varying activation conditions during cyclical length changes. A comparison to experimental data from two different muscle types suggests that the model can effectively predict the time course of force and mechanical energetic output of muscle for a wide range of contraction conditions, particularly during activation of the muscle. With fixed activation properties, discrepancies in the work output between the model and the experimental results were greatest at the faster and slower cycle frequencies than that for which the model was optimised. Further optimisation of the activation properties across each individual cycle frequency examined demonstrated that a change in the relationship between the concentration of the activator (Ca2+) and the activation level could account for these discrepancies. The variation in activation properties with speed provides evidence for the phenomenon of shortening deactivation, whereby at higher speeds of contraction the muscle deactivates at a faster rate. The results of this study demonstrate that predictions about the mechanics and energetics of muscle are possible when sufficient information is known about the specific muscle.

70 μM caffeine treatment enhances in vitro force and power output during cyclic activities in mouse extensor digitorum longus muscle

Caffeine ingestion by human athletes has been found to improve endurance performance primarily acting via the central nervous system as an adenosine receptor antagonist. However, a few studies have implied that the resultant micromolar levels of caffeine in blood plasma (70 microM maximum for humans) may directly affect skeletal muscle causing enhanced force production. In the present study, the effects of 70 microM caffeine on force and power output in isolated mouse extensor digitorum longus muscle were investigated in vitro at 35 degrees C. Muscle preparations were subjected to cyclical sinusoidal length changes with electrical stimulation conditions optimised to produce maximal work. 70 microM caffeine caused a small but significant increase (2-3%) in peak force and net work produced during work loops (where net work represents the work input required to lengthen the muscle subtracted from the work produced during shortening). However, these micromolar caffeine levels did not affect the overall pattern of fatigue or the pattern of recovery from fatigue. Our results suggest that the plasma concentrations found when caffeine is used to enhance athletic performance in human athletes might directly enhance force and power during brief but not prolonged activities. These findings potentially confirm previous in vivo studies, using humans, which implied caffeine ingestion may cause acute improvements in muscle force and power output but would not enhance endurance.

Predicting muscle force generation during fast-starts for the common carp Cyprinus carpio

Muscle contractile properties have been characterised for white myotomal muscle from the common carp Cyprinus carpio at 10, 15, and 20 °C. The time course of muscle force development was measured when one, two, or three stimuli were delivered at the onset of constant velocity shortening. As the shortening velocity increased several parameters decreased including the maximum force, the time course for the contraction and the relative duration of the deactivation compared to the activation. The maximum force and the relative rates of activation to deactivation for the contraction were relatively independent of temperature, whereas the duration of the contraction decreased with increasing temperature. A predictive model was developed which was based on fitting a modified Weibull distribution to these observations. The model was used to interpolate the expected contractile forces during cyclic length-changes. Measured and predicted values for force and power during such cyclic work-loop experiments showed an excellent agreement over the range of shortening regimes typically found during swimming behaviours. However, the predicted force was overestimated during the deactivation phase of the contractions when the shortening velocities exceeded those found during swimming.

Muscle contraction as a Markov process. I: Energetics of the process.

Force generation during muscle contraction can be understood in terms of cyclical length changes in segments of actin thin filaments moving through the three-dimensional lattice of myosin thick filaments. Recent anomalies discovered in connection with analysis of myosin step sizes in in vitro motility assays and with skinned fibres can be rationalized by assuming that ATP hydrolysis on actin accompanies these length changes. The paradoxically rapid regeneration of tension in quick release experiments, as well as classical energetic relationships, such as Hill's force-velocity curve, the Fenn effect, and the unexplained enthalpy of shortening, can be given mutually self-consistent explanations with this model. When muscle is viewed as a Markov process, the vectorial process of chemomechanical transduction can be understood in terms of lattice dependent transitions, wherein the phosphate release steps of the myosin and actin ATPases depend only on occurrence of allosteric changes in neighbouring molecules. Tropomyosin has a central role in coordinating the steady progression of these cooperative transitions along actin filaments and in gearing up the system in response to higher imposed loads.

Muscle contraction as a Markov process I: energetics of the process

Force generation during muscle contraction can be understood in terms of cyclical length changes in segments of actin thin filaments moving through the three-dimensional lattice of myosin thick filaments. Recent anomalies discovered in connection with analysis of myosin step sizes in in vitro motility assays and with skinned fibres can be rationalized by assuming that ATP hydrolysis on actin accompanies these length changes. The paradoxically rapid regeneration of tension in quick release experiments, as well as classical energetic relationships, such as Hill's force–velocity curve, the Fenn effect, and the unexplained enthalpy of shortening, can be given mutually self-consistent explanations with this model. When muscle is viewed as a Markov process, the vectorial process of chemomechanical transduction can be understood in terms of lattice dependent transitions, wherein the phosphate release steps of the myosin and actin ATPases depend only on occurrence of allosteric changes in neighbouring molecules. Tropomyosin has a central role in coordinating the steady progression of these cooperative transitions along actin filaments and in gearing up the system in response to higher imposed loads.

Optimal shortening velocities for in situ power production of rat soleus and plantaris muscles.

Force-velocity (FV) relationships have been used previously to calculate maximal power production and to identify an optimal velocity of shortening (V(opt)-fv) to produce such power in skeletal muscle. The cyclical nature of muscle position during locomotion for muscles such as the soleus and plantaris is such that either constant force or velocity is rarely attained. In the present study, the work loop technique, a technique developed to measure maximal attainable power output from muscles undergoing cyclic length changes, was undertaken to determine whether simulating in vivo function alters the power-velocity relationship of the soleus and plantaris and, in particular, the velocity of shortening that produces maximal power (V(opt)-wl). FV relationships were determined for both soleus (n = 4) and plantaris (n = 4) muscles in situ from adult female Sprague-Dawley rats by measuring shortening velocities during afterloaded isotonic contractions. The velocity that produced maximal power using FV relationships, V(opt)-fv, was 54.6 +/- 0.7 mm/s for the plantaris vs. 20.2 +/- 1.2 mm/s for the soleus. Then, the work loop technique was employed to measure net power from these same muscles at multiple cycling frequencies (1.5 to 4.0 Hz for the soleus; 4.0 to 8.0 Hz for the plantaris). Multiple power-velocity curves were generated (one at each cycle frequency) by varying the strain (1-8 mm). Thus, at each cycle frequency, V(opt)-wl could be identified. For both the plantaris and soleus, V(opt)-wl at each cycle frequency was not different from their respective V(opt)-fv value. Thus both fast and slow skeletal muscles have inherent optimal shortening velocities, identifiable with FV relationships, that dictate their respective maximal attainable mechanical power production using the work loop technique.

Interaction of the movement-dependent, extrafusal and fusimotor after-effects in the firing of the primary spindle endings

The after-effects of the firing of the primary spindle endings were studied in ankle extensor muscles of cats under Nembutal anesthesia. The activities of 27 primary endings of the muscle spindles from mm. soleus, plantaris, and gastrocnemius have been analysed in various combinations of fusimotor and extrafusal stimulation and application of the mechanostimulation to the spindle bearing muscle. Short-term simulation of static γ-axons evoked a post-stimulation increase in the spindle ending firing, which can be recorded under both isometric and isotonic conditions on applying a weak extrafusal stimulation or without it. The movement-dependent after-effects were tested with a double-trapezoid pattern of muscle length (or load) changes. The after-effects consisted of the difference of firing rates at the same values of muscle length (or load) with opposite direction of movement to the steady states; these uncertainties were also present during constant stimulation of static γ-axons. The rate difference showed a tendency to a certain decrease with stimulation rate increment. For diapason of the stimulation rates up to 125impulses/s a small negative correlation (r=−0.61) between the firing rate differences and the γ-stimulation rate has been registered in the population of primary endings tested under length servo-control conditions.Using a frequency-modulated intrafusal stimulation, a clockwise hysteresis dependence of the spindle firing rate upon stimulation rate was demonstrated. The pronounced after-effects were shown to exist for steady rates of stimulation: the discharge rates were always higher after stimulation rate increase and lower after its decrease. Fusimotor after-effects were effectively destroyed by both the extrafusal stimulation and the cyclic length (load) changes evoked lengthening–shortening movements of the muscle.The results obtained can be considered as evidence for a hypothesis that history-dependent behavior of muscle spindles is mainly connected with hysteresis of the intrafusal muscle fibers and the whole spindle bearing muscle.

Electromotility of outer hair cells from the cochlea of the echolocating bat, Carollia perspicillata

Isolated outer hair cells (OHCs) and explants ot the organ of Corti were obtained from the cochlea of the echolocating bat, Carollia perspicillata, whose hearing range extends up to about 100 kHz. The OHCs were about 10-30 microns long and produced resting potentials between -30 to -69 mV. During stimulation with a sinusoidal extracellular voltage field (voltage gradient of 2 mV/microns) cyclic length changes were observed in isolated OHCs. The displacements were most prominent at the level of the cell nucleus and the cuticular plate. In the organ of Corti explants, the extracellular electric field induced a radial movement of the cuticular plate which was observed using video subtraction and photodiode techniques. Maximum displacements of about 0.3-0.8 microns were elicited by stimulus frequencies below 100 Hz. The displacement amplitude decreased towards the noise level of about 10-30 nm for stimulus frequencies between 100-500 Hz, both in apical and basal explants. This compares well with data from the guinea pig, where OHC motility induced by extracellular electrical stimulation exhibits a low pass characteristic with a corner frequency below 1 kHz. The data indicate that fast OHC movements presumably are quite small at ultrasonic frequencies and it remains to be solved how they participate in amplifying and sharpening cochlear responses in vivo.

The influence of temperature on power output of scup red muscle during cyclical length changes.

To gain insight into how temperature affects locomotory performance, we measured power output of scup red muscle during oscillatory length changes at 10 degrees C and 20 degrees C. When we optimized work loop parameters, we found that maximum power was 27.9 W kg-1 at 20 degrees C and 12.8 W kg-1 at 10 degrees C, giving a Q10 of 2.29. Maximum power was generated at a higher oscillation frequency at 20 degrees C (5 Hz) than at 10 degrees C (2.5 Hz), and the Q10 for power output at a given oscillation frequency ranged from about 1 at 1 Hz to about 5 at 7.5 Hz. An analysis of the results in terms of crossbridge kinetics suggests that the higher power output at 20 degrees C is associated with both a higher Vmax (i.e. more power per cycling crossbridge) and faster activation and relaxation (i.e. a greater number of cycling crossbridges). To obtain a more realistic understanding of the functional importance of temperature effects on muscle, we imposed on isolated muscle in vivo length changes and oscillation frequencies (measured during previous experiments on swimming scup) and the in vivo stimulus duty cycle (measured from electromyograms in this study). At 20 degrees C, muscle bundles tested under these in vivo conditions produced nearly maximal power, suggesting that the muscle works optimally during locomotion and, thus, important contractile parameters have been adjusted to produce maximum mechanical power at the oscillation frequencies and length changes needed during swimming. At 10 degrees C, muscle bundles tested under in vivo conditions produced much less power than was obtained during the 'optimized' work loop experiments discussed above. Furthermore, although 'optimized' work loop experiments showed that maximum power output occurs at 2.5 Hz, scup do not appear to swim with such a low tailbeat frequency. Although the reason for these apparent discrepancies at 10 degrees C are not known, it shows that simple extrapolation from isolated muscle to the whole animal, without knowledge of how the muscle is actually used in vivo, can be misleading.

Efferent and afferent activity in a gastrocnemius nerve branch during locomotion in the thalamic cat.

The firing patterns of alpha and gamma efferent fibres and of group I and group II afferent fibres innervating the gastrocnemius muscle were observed during spontaneous locomotor movements in the thalamic cat. Multi-unit discharges of each kind of fibre were obtained by electronic sorting of the action potentials from the whole activity of a thin branch of gastrocnemius lateralis or medialis nerve. The main results were: During the locomotor cycle the activity of the afferent and efferent populations was highly modulated. alpha- and gamma-motoneurones were co-activated within the locomotor cycle during ankle plantar-flexion. The gamma discharge began to rise earlier and to fall later than did the alpha discharge. The amplitude of the gamma discharge, unlike that of the alpha discharge, was largely independent of the vigour of walking. Between the cyclic discharges, most of gamma populations were tonically active whereas alpha populations were silent. Subgroups of the alpha and gamma populations were not usually activated according to the cell-size principle, but, the activation of the latest gamma subgroup always preceded that of the earliest alpha subgroup. Modulation of the group I and II afferent discharges was closely related to the cyclic length changes of the parent muscle. Fusimotor activation during the active shortening of gastrocnemius muscle prevented the afferent discharges from pausing. The pattern of afferent and efferent activity during selective curarisation of the extrafusal junctions indicated that the discharge of static gamma-motoneurones is modulated during the locomotor cycle. After curarisation of both extrafusal and intrafusal junctions, an efferent-discharge pattern of central origin persisted alternately in extensor- and flexor-muscle nerves (fictive locomotion). The durations of the fictive locomotor cycle and of the cyclic discharge in the sartorius nerve were increased as a consequence of the suppression of phasic afferent inputs to the C.N.S. Maintained ankle dorsi-flexion slowed the fictive locomotor rhythm and elicited opposite effects, respectively excitation and depression, on the magnitude of the alpha and gamma discharges. Maintained ankle plantar-flexion scarcely perturbed the duration of the fictive locomotor cycle, but the duration of the sartorius-nerve discharge lengthened at the expense of that of the gastrocnemius discharge. Both gastrocnemius alpha- and gamma-motoneurones were depressed, the former considerably more than the latter. The roles of the gastrocnemius afferents and gamma-efferents during the locomotor cycle are discussed in the light of these results.

Cyclical cell length changes in wood in relation to storied structure and interlocked grain

Interlocked grain can result from migration of a succession of left (S) and right (Z) orientational domains along the cambium. This grain pattern occurs both in species having nonstoried and those having storied cambium. In storied cambia, storey height being fixed, geometry requires that cambial fusiform initial cells undergo cyclical length changes in phase with cyclical inclination changes, e.g., a 4% length increase for a 16° inclination. We can expect the length changes to be detectable in wood if vessel member length reflects fusiform initial cell length and if the absolute amount of intrusive growth during differentiation of fibers is nearly invariant. Measurements of cells from maximum S, maximum Z, and axially aligned grain (I) sites along a radius in Entandrophragma cyclindricum and E. utile wood (storied) revealed clearly cyclical length changes in vessel members, parenchyma strands, and xylem fibers. Measurements in Nyssa sylvatica and Platanus acerifolia (nonstoried) revealed no such changes. Hence in species having storied cambium, cyclical changes in length of wood cells can accompany the slow endogenous rhythm manifested by interlocked grain. This is true because intrusive growth in these species, though extensive, does not obi iterate effects of small differences in cambial initial cell length.