Mice were anaesthetized with isoflurane and placed supine on a platform heated via circulating water bath at 37°C. The skin was removed from the right leg exposing the TA muscle, and the lower ligament was tied with a looped suture at the head of the TA. The patella was secured to the heated platform with a 25‐G needle, and the looped suture was fixed to the Dual Mode Muscle Lever (300‐C, Aurora Scientific Inc.). Two electrodes were placed above the tibia and under the TA. A 1 Hz electrical stimulation was delivered (0.2 s duration, 1 s between stimulations) via stimulator (701C, Aurora Scientific Inc.), and tension of the TA was increased until an optimal tension was achieved (Lo). After optimal tension was established, a force frequency of isometric contractions was initiated (0.2 s duration, 120 s between stimulations). All data were collected and analysed using the manufacturer supplied software (DMC and DMA, Aurora Scientific Inc.). Mice were housed at thermoneutral temperature prior to this measurement.
Dual mode muscle lever
Manufactured by Aurora Scientific
Sourced in Canada
The Dual-mode muscle lever is a laboratory instrument designed to study the mechanical properties of muscle tissue. It is capable of operating in both force and length control modes, allowing researchers to precisely measure the force generated by a muscle sample or control its length. The device features a high-resolution force transducer and linear actuator to facilitate accurate data collection during experiments.
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3 protocols using dual mode muscle lever
1
Isometric Muscle Tension Measurement
2
Isometric Muscle Force Measurement
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Mice were anesthetized by IP injection with a mixture of 10 mg/ml dolethal (Vetoquinol, UK) and 15 μg/ml buprenodale (Dechra, UK) at five times of the bodyweight. The distal tendon of tibialis anterior (TA) muscle was dissected and attached to an isometric transducer, Dual-mode muscle lever (Aurora Scientific, Canada), through a loop made of braided silk suture (Harvard Apparatus, UK). The sciatic nerve was isolated and distally stimulated by a bipolar silver electrode using supramaximal square wave pulses at 0.1 ms duration. Data provided by the isometric transducer were recorded and analyzed using Dynamic Muscle Control and Analysis Software (Aurora Scientific, Canada). All isometric measurements were obtained at an initial length at which a maximal tension was recorded during the tetanus. Responses to tetanic stimulations at increased pulse frequencies from 10 to 180 Hz were recorded and the maximal force (mN) was determined. The specific force (mN/mm2) was subsequently calculated based on a ratio of the maximal force and the muscle cross-sectional area (CSA) that was approximated mathematically by dividing the muscle mass by the optimum fiber length and the density of mammalian muscle, as described in TREAT-NMD SOP DMD_M.2.2.005.
3
Measuring Muscle Contractile Function in Mice
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Mice were anesthetized by IP injection with a mixture of 10 mg/ml dolethal (Vetoquinol, UK) and 15 µg/ml buprenodale (Dechra, UK) at 5 times of the bodyweight. The distal tendon of tibialis anterior (TA) muscle was dissected and attached to an isometric transducer, Dual-mode muscle lever (Aurora Scientific, Canada), through a loop made of braided silk suture (Harvard Apparatus, UK). The sciatic nerve was isolated and distally stimulated by a bipolar silver electrode using supramaximal square wave pulses at 0.1 ms duration. Data provided by the isometric transducer were recorded and analyzed using Dynamic Muscle Control and Analysis Software (Aurora Scientific, Canada). All isometric measurements were obtained at an initial length at which a maximal tension was recorded during the tetanus. Responses to tetanic stimulations at increased pulse frequencies from 10 Hz to 180 Hz were recorded and the maximal force (mN) was determined. The specific force (mN/mm 2 ) was subsequently calculated based on a ratio of the maximal force and the muscle cross-sectional area (CSA) that was approximated mathematically by dividing the muscle mass by the optimum fiber length and the density of mammalian muscle, as described in TREAT-NMD SOP DMD_M.2.2.005. Force measurement was performed in a blinded manner.
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