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.

Achilles tendon (AT) comprises of 3 subtendons arising from the soleus (SOL) and the lateral (LG) and medial (MG) heads of the gastrocnemius muscle. While recent human studies show differential displacement within AT, these displacements have not been attributed to specific subtendons. We tested the hypothesis that the SOL and LG subtendons show differential displacement and strain during various combinations of SOL, LG, and MG excitations. Movement of knots, sutured onto SOL and LG subtendons of 12 Wistar rats, was videotaped, while the muscles were stimulated intramuscularly and ankle torque was assessed. When SOL only was stimulated, the plantar flexion torque was the smallest among the different conditions (P < .001). In this condition, from passive to active state, the displacement (0.57 vs 0.47 mm, P = .002) and strain (8.4% vs 2.4%, P < .001) in the SOL subtendon were greater than in LG subtendon. When LG only was stimulated, a higher ankle torque was measured as compared to SOL stimulation (P < .001); the displacement was similar in both subtendons (~0.6 mm), while the strain was greater in LG than in SOL (4.7% vs 1.7%, P < .001). When all 3 muscles were stimulated simultaneously, ankle torque was highest and the displacement (0.79 vs 0.74 mm, P = .002) and strain (7.7% vs 4.4%, P = .003) were greater in SOL than in LG. These data show that the different subtendons of AT can experience relative displacement and differential strains. Together with anatomical dissections, the results revealed that such uniformities may be due to a lower stiffness of SOL subtendon compared to LG.

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.

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.

To relate in vivo behavior of fascicle segments within a muscle to their in vitro force-length relationships, we examined the strain behavior of paired segments within each of three vertebrate muscles. After determining in vivo muscle activity patterns and length changes of in-series segments within the semimembranosus muscle (SM) in the American Toad (Bufo americanus) during hopping and within the sternohyoid (SH) muscle in the rat (Rattus rattus) during swallowing, and of spatially separated fascicles within the medial gastrocnemius (MG) muscle in the rat during trotting, we measured their corresponding in vitro (toad) or in situ (rat) force-length relationships (FLRs). For all three muscles, in vivo strain heterogeneity lasted for about 36-57% of the behavior cycle, during which one segment or fascicle shortened while the other segment or fascicle simultaneously lengthened. In the toad SM, the proximal segment shortened from the descending limb across the plateau of its FLR from 1.12 to 0.91 of its optimal length (Lo), while the distal segment lengthened (by 0.04 ± 0.04 Lo) before shortening down the ascending limb from 0.94 to 0.83 Lo. In the rat SH muscle, the proximal segment tended to shorten on its ascending limb from 0.90 to 0.85 Lo while the distal segment tended to lengthen across Lo (0.96-1.12 Lo). In the rat MG muscle, in vivo strains of proximal fascicles ranged from 0.72 to 1.02 Lo, while the distal fascicles ranged from 0.88 to 1.11 Lo. Even though the timing of muscle activation patterns were similar between segments, the heterogeneous strain patterns of fascicle segments measured in vivo coincided with different operating ranges across their FLRs simultaneously, implying differences in force-velocity behavior as well. The three vertebrate skeletal muscles represent a diversity of fiber architectures and functions and suggest that patterns of in vivo contractile strain and the operating range over the FLR in one muscle region does not necessarily represent other regions within the same muscle.

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.

a b s t r a c tSkeletal 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 ¼ 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 ( þ1.1 7 1.1% mean7 s.d.) and frontal plane direction. The latter was affected by knee angle and ranged from þ 0.27 0.3°at 60°to þ1.1 7 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.

Skeletal muscle has fiber architectures ranging from simple to complex, alongside variation 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 relations 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±0.8mm vs 10.9±0.7mm) fibers and steeper (28.7±11.0° vs 19.6±6.3°) fiber angles at optimum length, and operates over a larger (17.9 ± 3.8% vs 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 vs 2.4 ± 1.5°/mm muscle shortening) and architectural gearing (1.82 ± 0.53 vs 1.25 ± 0.24); whereas, the distal region exhibits greater peak shortening speed (96.0mm/s vs 81.3mm/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.