Force Of Extensor Digitorum Longus Muscle Research Articles

The Implication of Hinge 1 and Hinge 4 in Micro-Dystrophin Gene Therapy for Duchenne Muscular Dystrophy.

Duchenne muscular dystrophy (DMD) is a fatal muscle disease caused by dystrophin deficiency. Dystrophin consists of the amino terminus, central rod domain with 24 spectrin-like repeats and four hinges (H), cysteine-rich domain, and carboxyl terminus. Several highly abbreviated micro-dystrophins (μDys) are currently in clinical trials. They all carry H1 and H4. In this study, we investigated whether these two hinges are essential for μDy function in murine DMD models. Three otherwise identical μDys were engineered to contain H1 and/or H4 and were named H1/H4 (with both H1 and H4), ΔH1 (without H1), and ΔH4 (without H4). These constructs were packaged in adeno-associated virus serotype-9 and delivered to the tibialis anterior muscle of 3-month-old male mdx4cv mice (1E12 vector genome particles/muscle). Three months later, we detected equivalent μDys expression in total muscle lysate. However, only H1/H4 and ΔH1 showed correct sarcolemmal localization. ΔH4 mainly existed as sarcoplasmic aggregates. H1/H4 and ΔH1, but not ΔH4, fully restored the dystrophin-associated protein complex and significantly improved the specific muscle force. Eccentric contraction-induced force decline was best protected by H1/H4, followed by ΔH1, but not by ΔH4. Next, we compared H1/H4 and ΔH1 in 6-week-old male mdx mice by intravenous injection (1E13 vector genome particles/mouse). Four months postinjection, H1/H4 significantly outperformed ΔH1 in extensor digitorum longus muscle force measurements but two constructs yielded comparable electrocardiography improvements. We conclude that H4 is essential for μDys function and H1 facilitates force production. Our findings will help develop next-generation μDys gene therapy.

The Effect of Short Nerve Grafts in Series on Axonal Regeneration Across Isografts or Acellular Nerve Allografts

To evaluate the regenerative effect of the additional suture line when using either isografts (ISOs) or acellular nerve allografts (ANAs) placed end-to-end to span a short gap in a rat model. Rat sciatic nerves were transected and repaired with 2-cm nerve grafts (ISO or ANA). The grafts were 2 cm in length or a 1-cm segment was connected end-to-end to a 1-cm segment to yield a 2-cm length. At 8 weeks, extensor digitorum longus (EDL) muscle force and mass were measured. Nerves were harvested for histomorphometry. In a separate parallel study, the nerves were harvested 2 weeks following graft implantation to assess gene expression changes. All grafts demonstrated regeneration across the 2-cm segment(s). The additional suture line did not result in statistical differences in the number of myelinated nerve fibers that reached the distal nerve. However, when the graft types were compared, there was a significant decrease in nerve fibers in the ANA groups. The EDL muscle mass was significantly greater by using nerve ISOs compared with ANAs, regardless of an additional suture line, but there were no statistical differences noted in EDL muscle force. Gene expression analysis did not differ owing to an additional suture line. Minimal axonal loss and no functional deficits were identified with an additional suture line in this rodent short nerve gap model. Placing nerve grafts in series is a viable option for treating short nerve gaps; however, the use of autografts remains preferable over the use of ANAs.

The Angiotensin Converting Enzyme Inhibitor Lisinopril Improves Muscle Histopathology but not Contractile Function in a Mouse Model of Duchenne Muscular Dystrophy.

Background: Angiotensin converting enzyme inhibitors (ACEi) are the current standard of care treatment for cardiac dysfunction in Duchenne muscular dystrophy patients. We previously showed treatment with an ACEi plus mineralocorticoid receptor (MR) antagonist improves limb and respiratory skeletal muscles, in addition to cardiac muscles, in a dystrophic mouse model at 20 weeks-of-age. Objective: To determine whether previously observed preclinical benefits of an ACEi plus MR antagonist on dystrophic skeletal muscles can be reproduced by increasing ACEi dosage alone. We also compared functional and histological outcome measures at 10 and 20 weeks-of-age. Methods: Dystrophin deficient utrophin haplo-insufficient (utrn +/- ; mdx) “het” mice were treated with 10, 20, or 50 mg/kg × day of the ACEi lisinopril from 4 to 10 weeks-of-age via water bottles and compared with C57BL/10 wild-type control mice and untreated hets. Data from 10 week-old het mice were also compared to data collected from an untreated het group at 20 weeks-old. In vivo cardiac and grip strength measurements, in vitro diaphragm and extensor digitorum longus muscle force measurements, and histopathological analyses were performed. One-way ANOVA followed by Dunnett post hoc comparison was used to determine significance. Results: ACEi treatment reduced skeletal muscle damage but had no significant effect on muscle force. Body weight, heart rate, grip strength and blood pressure were unaffected by treatment. Limb muscle histopathology was more informative at 10 than 20 weeks-of-age.Conclusions: These results suggest increased ACEi dosage alone cannot improve all dystrophic parameters. Further optimization of MR antagonists in 20 week-old mice is warranted.

Myofascial force transmission is increasingly important at lower forces: firing frequency‐related length–force characteristics of rat extensor digitorum longus

Effects of submaximal stimulation frequencies on myofascial force transmission were investigated for rat anterior crural muscles with all motor units activated. Tibialis anterior and extensor hallucis longus (TAEHL) muscles were kept at constant muscle-tendon complex length, but extensor digitorum longus muscle (EDL) was lengthened distally. All muscles were activated simultaneously at 10, 20, 30 and 100 Hz within an intact anterior crural compartment. At lower frequencies, significant proximo-distal EDL force differences exist. Absolute EDL proximo-distal active force differences were highest at 100 Hz (deltaF(dist-prox) = 0.4 N). However, the normalized difference was highest at 10 Hz (deltaF(dist-prox) = 30%F(dist)). Firing-frequency dependent shifts of the ascending limb of the EDL length-force curve to higher lengths were confirmed for a muscle within an intact compartment, although effects of firing frequency assessed at proximal and distal EDL tendons differed quantitatively. As EDL was lengthened distally, TAEHL distal isometric active force decreased progressively. The absolute decrease was highest for 100 Hz (deltaF(from initial) = -0.25 N). However, the highest normalized decrease was found for 10 Hz stimulation (deltaF(from initial) = -40%). At submaximal stimulation frequencies, myofascial force transmission is present and the fraction of force transmitted myofascially increases with progressively lower firing frequencies. Evidently, the stiffness of epimuscular myofascial paths of force transmission decreases less than the stiffness of serial sarcomeres and myotendinous pathways. It is concluded that low firing frequencies as encountered in vivo enhance the relative importance of epimuscular myofascial force transmission with respect to myotendinous force transmission.

Non-Myotendinous Force Transmission in Rat Extensor Digitorum Longus Muscle

The extensor digitorum longus muscle (EDL) of the rat hindleg consists of four heads. The heads are named after their insertions on the digits of toes II, III, IV and V. The EDL heads share a proximal tendon and aponeurosis, but have separate distal aponeuroses and tendons. By cutting the distal tendons of selected heads, direct myotendinous force transmission within these heads is prevented. Therefore, force exerted by the muscle would be expected to decrease according to the physiological cross-sectional area disconnected if myotendinous force transmission were the only mechanism of force transmission. The results indicate that EDL force production remained at high levels after acute tenotomy: muscle length-force curves did not alter significantly following cutting of the tendons of heads II and III. Cutting the tendon of head IV as well leaves only head V in its original condition. After tenotomy of head IV, length-force characteristics were altered significantly, but optimum force was maintained at 84% of that of the intact muscle. After separation of head IV from head V intramuscularly for some distance along their interface, the force dropped to much lower levels, with optimum force approaching 50% of that of the intact muscle. The length of active proximal fibres (located within head II) did not remain constant but increased with increasing muscle lengths after tenotomy as well as after partial separation of heads IV and V. The amount of length change decreased after intramuscular separation of the heads, indicating declining reactive forces. It is concluded that force transmission occurred from tenotomized heads to their intact neighbours and vice versa. The magnitude of the force transmitted from head to head was dependent on the degree of integrity of the connective tissue at the interface between heads.

Tubular Nerve Guide and Epineurial Repair: Comparison of Techniques for Neurorrhaphy

Experiments were conducted on 36 male, Sprague-Dawley rats. In 10 animals, neurorrhaphy was performed on the peroneal nerve with epineurial repair and, in 11 animals, with a tubular polyethylene nerve guide. The authors tested the hypothesis that, following transient denervation of a skeletal muscle by transection of a peroneal nerve, the restoration of maximum force and of maximum specific force developed after insertion of a tubular nerve guide, will not be different from that developed after microsurgical epineurial neurorrhaphy. The contractile properties of the extensor digitorum longus (EDL) muscle, innervated by the peroneal nerve, were evaluated after an average of 116 days. The maximum tetanic force of EDL muscles with epineurial repair and nerve guide were 84 percent and 75 percent, respectively, of the value for control EDL muscles. The specific forces of the muscles in both groups were not different from the control values. The conclusion is that, following stabilization after transection and repair, each of the two methods was equally effective in restoring the ability of the muscle to develop force.