Effect of muscle stimulation intensity on the heterogeneous function of regions within an architecturally complex muscle.

Skeletal muscle has fiber architectures ranging from simple to complex, alongside variations in fiber-type and neuro-anatomical compartmentalization. However, the functional implications of muscle subdivision into discrete functional units remain poorly understood. The rat medial gastrocnemius has well-characterized regions with distinct architectures and fiber type composition. Here, force-length and force-velocity contractions were performed for two stimulation intensities (supramaximal and submaximal) and for three structural units (whole muscle belly, proximal region, and distal region) to assess the effect of muscle compartmentalization on contractile force-length-velocity relationships and optimal speed for power production. Additionally, fiber strain, fiber rotation, pennation, and architectural gearing were quantified. Our results suggest that the proximal and distal muscle regions have fundamentally different physiological function. During supramaximal activation, the proximal region has shorter (8.4 ± 0.8 mm versus 10.9 ± 0.7 mm) fibers and steeper (28.7 ± 11.0° versus 19.6 ± 6.3°) fiber angles at optimum length, and operates over a larger (17.9 ± 3.8% versus 12.6 ± 2.7%) range of its force-length curve. The proximal region also exhibits larger changes in pennation angle (5.6 ± 2.2°/mm versus 2.4 ± 1.5°/mm muscle shortening) and architectural gearing (1.82 ± 0.53 versus 1.25 ± 0.24), whereas the distal region exhibits greater peak shortening speed (96.0 mm/s versus 81.3 mm/s) and 18-27% greater optimal speed. Overall, similar patterns were observed during submaximal activation. These regional differences in physiological function with respect to the whole muscle highlight how variation in motor recruitment could fundamentally shift regional functional patterns within a single muscle, which likely has important implications for whole muscle force and work output in vivo.NEW & NOTEWORTHY We show that muscle compartmentalization can influence whole muscle contractile properties, with slower-fibered proximal rat medial gastrocnemius undergoing larger changes in pennation angle and architectural gearing, whereas the faster-fibered distal region achieves greater peak and optimal shortening velocity, and power output. Consequently, regional variation in motor recruitment can fundamentally influence functional patterns within a single muscle.

In vivo force-length and activation dynamics of two distal rat hindlimb muscles in relation to gait and grade.

Muscle function changes to meet the varying mechanical demands of locomotion across different gait and grade conditions. A muscle's work output is determined by time-varying patterns of neuromuscular activation, muscle force and muscle length change, but how these patterns change under different conditions in small animals is not well defined. Here, we report the first integrated in vivo force-length and activation patterns in rats, a commonly used small animal model, to evaluate the dynamics of two distal hindlimb muscles (medial gastrocnemius and plantaris) across a range of gait (walk, trot and gallop) and grade (level and incline) conditions. We use these data to explore how the pattern of force production, muscle activation and muscle length changes across conditions in a small quadrupedal mammal. As hypothesized, we found that the rat muscles show limited fascicle strains during active force generation in stance across gaits and grades, indicating that these distal rat muscles generate force economically but perform little work, similar to patterns observed in larger animals during level locomotion. Additionally, given differences in fiber type composition and variation in motor unit recruitment across the gait and grade conditions examined here for these muscles, the in vivo force-length behavior and neuromuscular activation data reported here can be used to validate improved two-element Hill-type muscle models.

Myofascial Loads Can Occur without Fascicle Length Changes.

Many studies have shown that connective tissue linkages can transmit force between synergistic muscles and that such force transmission depends on the position of these muscles relative to each other and on properties of their intermuscular connective tissues. Moving neighboring muscles has been reported to cause longitudinal deformations within passive muscles held at a constant muscle-tendon unit (MTU) length (e.g., soleus [SO]), but muscle forces were not directly measured. Deformations do not provide a direct measure of the force transmitted between muscles. We combined two different muscle preparations to assess whether myofascial loads exerted by neighboring muscles result in length changes of SO fascicles. We investigated the effects of proximal MTU length changes of two-joint gastrocnemius (GA) and plantaris (PL) muscles on the fascicle length of the one-joint SO muscle within (1) an intact muscle compartment and (2) a disrupted compartment that allowed measurements of fascicle length and distal tendon force of SO simultaneously. SO muscle bellies of Wistar rats (n = 5) were implanted with sonomicrometry crystals. In three animals, connectivity between SO and GA+PL was enhanced. Measurements were performed before and during maximal excitation of all plantar flexor muscles. In both setups, MTU length of GA+PL did not affect the length of SO fascicles, neither during passive nor active conditions. However, lengthening the MTU of GA+PL increased distal tendon force of SO by 43.3-97.8% (P < 0.001) and 27.5-182.6% (P < 0.001), respectively. This indicates that substantial myofascial force transmission between SO and synergistic muscle can occur via a connective tissue network running parallel to the series of SO sarcomeres without substantial length changes of SO fascicles.

Synergistic Co-activation Increases the Extent of Mechanical Interaction between Rat Ankle Plantar-Flexors.

Force transmission between rat ankle plantar-flexors has been found for physiological muscle lengths and relative positions, but only with all muscles maximally activated. The aims of this study were to assess intermuscular mechanical interactions between ankle plantar-flexors during (i) fully passive conditions, (ii) excitation of soleus (SO), (iii) excitation of lateral gastrocnemius (LG), and (iv) during co-activation of SO, and LG (SO&LG). We assessed effects of proximal lengthening of LG and plantaris (PL) muscles (i.e., simulating knee extension) on forces exerted at the distal SO tendon (FSO) and on the force difference between the proximal and distal LG+PL tendons (ΔFLG+PL) of the rat. LG+PL lengthening increased FSO to a larger extent (p = 0.017) during LG excitation (0.0026 N/mm) than during fully passive conditions (0.0009 N/mm). Changes in FSO in response to LG+PL lengthening were lower (p = 0.002) during SO only excitation (0.0056 N/mm) than during SO&LG excitation (0.0101 N/mm). LG+PL lengthening changed ΔFLG+PL to a larger extent (p = 0.007) during SO excitation (0.0211 N/mm) than during fully passive conditions (0.0157 N/mm). In contrast, changes in ΔFLG+PL in response to LG+PL lengthening during LG excitation (0.0331 N/mm) were similar (p = 0.161) to that during SO&LG excitation (0.0370 N/mm). In all conditions, changes of FSO were lower than those of ΔFLG+PL. This indicates that muscle forces were transmitted not only between LG+PL and SO, but also between LG+PL and other surrounding structures. In addition, epimuscular myofascial force transmission between rat ankle plantar-flexors was enhanced by muscle activation. However, the magnitude of this interaction was limited.

Limited mechanical effects of intermuscular myofascial connections within the intact rat anterior crural compartment.

Skeletal muscles of the rat anterior crural compartment are mechanically connected by epimuscular myofascial connections, but the relevance for mechanical muscle function within physiological ranges of joint motion is unclear. We evaluated the net effect at the ankle joint of epimuscular myofascial connections between tibialis anterior (TA) and extensor digitorum longus (EDL) muscles in the rat (n=8) and determined which anatomical structures may mediate such epimuscular mechanical interactions. We assessed (1) effects of knee angle (i.e. changes in EDL length and position relative to TA) and interactions of knee angle with fasciotomy and proximal EDL tenotomy on TA ankle moment and (2) the effect of knee angle on TA and EDL ankle moment summation. Knee angle was varied between 60° and 130°. Ankle angle was kept constant (90°). TA and EDL were excited individually and simultaneously (TA&EDL). The mathematical sum of individual TA and EDL moments was compared with the moment exerted by TA&EDL to assess the extent of non-additive ankle moment summation. Magnitude of TA ankle moment was not affected by knee angle, but frontal plane moment direction was. However, dissections indicated that this was not caused by the compartmental fascia or EDL length changes. Moment summation was non-additive in magnitude (+1.1±1.1% mean±s.d.) and frontal plane direction. The latter was affected by knee angle and ranged from +0.2±0.3° at 60° to +1.1±0.6° at 130°. As the net effects found were very limited, we conclude that myofascial connections between muscles in the anterior crural compartment have limited mechanical relevance during normal movement.

Effects of epimuscular myofascial force transmission on sarcomere length of passive muscles in the rat hindlimb.

Results from imaging studies and finite element models suggest epimuscular myofascial effects on sarcomere lengths in series within muscle fibers. However, experimental evidence is lacking. We evaluated epimuscular myofascial effects on (1) muscle belly, fiber, and mean sarcomere length and (2) sarcomere length distribution within passive fibers of the rat tibialis anterior (TA) and soleus (SO) muscles. Hindlimbs (n = 24) were positioned in predefined knee (55°, 90°, 125°, 160°) and ankle (either 90° or 125°) angles, and fixed in a formaldehyde solution. Varying knee joint angle causes changes in muscle-tendon unit length of SO and TA's synergists, but not of SO and TA. Whole fibers were taken from SO and TA and photographed along their length. Mean sarcomere length was assessed for the entire fiber and for the proximal, intermediate, and distal thirds (fiber segments) separately. Mean sarcomere length of the fiber was not affected by knee angle, neither for SO (mean: 2.44 ± 0.03 µm and 2.19 ± 0.05 µm for ankle angles of 90° and 125°, respectively) nor for TA (mean: 2.33 ± 0.05 µm and 2.51 ± 0.07 µm for ankle angle set to 90° and 125°, respectively). Only for TA, a significant interaction between knee angle and fiber segment was found, indicating changes in the distribution of lengths of in-series sarcomeres. Thus, while epimuscular myofascial force transmission did not cause mean sarcomere length changes within passive SO and TA, it did alter the length distribution of sarcomeres within passive TA.

No functionally relevant mechanical effects of epimuscular myofascial connections between rat ankle plantar flexors.

Triceps surae muscles are mechanically connected by the shared Achilles tendon and by epimuscular myofascial connections. We aimed to assess the effects of proximal lengthening of gastrocnemius and plantaris muscles, imposed by changes in knee angle, on the magnitude and direction of the 3D ankle moment exerted by the soleus muscle and the mechanical interaction between ankle plantar flexor muscles during co-activation of gastrocnemius muscle, in the rat (N=9). Ankle angle was kept constant (90 deg), whereas knee angle was varied between 60 deg and 130 deg. At each knee angle, the soleus muscle was excited individually as well as simultaneously with the gastrocnemius muscle (SO+GA). The mathematical sum of individual soleus and gastrocnemius ankle moments was compared with the ankle moment exerted by SO+GA to assess nonlinear summation. Knee angle did not affect the magnitude of the soleus ankle moment (P=0.695). Moment directions in the transverse (P=0.050) and frontal (P=0.008) planes were affected by knee angle, but dissection indicated that this was not caused by length changes of the two-joint synergistic muscles. Nonlinear summation was found in the magnitude (-1.4 ± 1.9%, mean ± s.d., P<0.001) and in the frontal plane vector direction of the ankle moment (0.13 ± 0.23 deg, P=0.003); however, the extent did not change with knee angle. Although contraction of SO+GA increased the length of the Achilles tendon compared with its length at rest, this was not dependent on the knee angle (P=0.649). Despite the fact that intermuscular force transmission per se cannot be excluded, we conclude that, in vivo, the mechanical effects of epimuscular myofascial connections between rat ankle plantar flexors are not functionally relevant.

Three-dimensional ankle moments and nonlinear summation of rat triceps surae muscles.

The Achilles tendon and epimuscular connective tissues mechanically link the triceps surae muscles. These pathways may cause joint moments exerted by each muscle individually not to sum linearly, both in magnitude and direction. The aims were (i) to assess effects of sagittal plane ankle angle (varied between 150° and 70°) on isometric ankle moments, in both magnitude and direction, exerted by active rat triceps surae muscles, (ii) to assess ankle moment summation between those muscles for a range of ankle angles and (iii) to assess effects of sagittal plane ankle angle and muscle activation on Achilles tendon length. At each ankle angle, soleus (SO) and gastrocnemius (GA) muscles were first excited separately to assess ankle-angle moment characteristics and subsequently both muscles were excited simultaneously to investigate moment summation. The magnitude of ankle moment exerted by SO and GA, the SO direction in the transverse and sagittal planes, and the GA direction in the transverse plane were significantly affected by ankle angle. SO moment direction in the frontal and sagittal planes were significantly different from that of GA. Nonlinear magnitude summation varied between 0.6±2.9% and -3.6±2.9%, while the nonlinear direction summation varied between 0.3±0.4° and -0.4±0.7° in the transverse plane, between 0.5±0.4° and 0.1±0.4° in the frontal plane, and between 3.0±7.9° and 0.3±2.3° in the sagittal plane. Changes in tendon length caused by SO contraction were significantly lower than those during contraction of GA and GA+SO simultaneously. Thus, moments exerted by GA and SO sum nonlinearly both in the magnitude and direction. The limited degree of nonlinear summation may be explained by different mechanisms acting in opposite directions.