In Vivo Assessment of Mouse Hindlimb Muscle Force, Contractile, and Fatigue Characteristics, and Motor Unit Number
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- Abstract
- Table of Contents
- Materials
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- Literature Cited
Abstract
The use of rodents to model neuromuscular diseases necessitates assessment of neuromuscular function to monitor disease progression. Muscle function can be assessed by determining muscle force, and the contraction's contractile and fatigue characteristics. Assessment of motor units gives a measure of motoneuron health. Thus, assessment of these parameters can reveal the degree and nature of neuromuscular pathology. A reduction in muscle force may result either from loss of motoneurons and a concomitant denervation of muscles or as a result of primary muscle pathology. Estimation of the number of functional motor units may identify whether the deficit is neural in origin. Here, we give a detailed description of the assessment of muscle force, contractile characteristics, and muscle fatigue, as well as a method that gives a direct and accurate readout on the number of motor units in individual mouse hindlimb muscles in mice?now widely used to model a variety of neuromuscular disorders. Curr. Protoc. Mouse Biol. 2:89?101 © 2012 by John Wiley & Sons, Inc.
Keywords: muscle force; muscle fatigue; muscle contraction; motor unit; motoneuron; twitch force
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PDF or HTML at Wiley Online LibraryTable of Contents
- Introduction
- Basic Protocol 1: Surgical Preparation of Mouse Hindlimb Muscles for Physiological Recordings
- Basic Protocol 2: Physiological Recording of Single‐Twitch Contractions to Determine Single‐Twitch Force and Time Characteristics of Single‐Twitch Contractions in Mouse Muscles
- Basic Protocol 3: Physiological Recording of Motor Units in the EDL Muscle of the Mouse
- Commentary
- Literature Cited
- Figures
- Tables
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Basic Protocol 1: Surgical Preparation of Mouse Hindlimb Muscles for Physiological Recordings
Materials
Basic Protocol 2: Physiological Recording of Single‐Twitch Contractions to Determine Single‐Twitch Force and Time Characteristics of Single‐Twitch Contractions in Mouse Muscles
Materials
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Figure 1. A graphic illustration on the position of muscles investigated in this protocol. The tibialis anterior (TA) and extensor digitorum longus (EDL) muscles are located anterior from the tibia, whereas the soleus muscle is located posterior from the bone; therefore, the dissection of this muscle is more practical from the prone position. View Image -
Figure 2. Analysis of single‐twitch traces obtained during isometric tension recordings. The figure shows two single‐twitch recordings obtained from the tibialis anterior (TA) muscles of a normal mouse (red represents the left side and blue represents right side). Using single‐twitch recordings like these, the following parameters can be read: twitch force ( g ) can be read by measuring the peak of the twitch trace; time to peak (TTP, milliseconds) is the time from the start of contraction until the peak of contraction; half relaxation time (HRT, milliseconds) is the time between the peak force and half value of the peak force on the relaxation phase. View Image -
Figure 3. Analysis of tetanic tension traces obtained during isometric tension recordings. The figure shows two tetanic tension traces obtained from the tibialis anterior (TA) muscles of a normal mouse (red represents the left side and blue represents right side). Single‐twitch recordings are also shown on the same graph. In order to achieve maximum force production the tetanic contractions were elicited by increasing stimulation frequencies; 40, 80, and 100 Hz. From recordings like these, the maximum force production can be measured in response to tetanic stimuli. Tetanic force ( g ) can be read by measuring the peak of the tetanus trace. View Image -
Figure 4. A characteristic fatigue trace obtained from a normal EDL muscle. The muscle was stimulated for 3 min through the sciatic nerve using 250‐msec tetanic pulses at 40‐Hz frequency. Muscle contractions were recorded using a chart recorder. For the analysis of the fatigue recording, the maximal and the minimal contraction trace were measured at the beginning and at the end of the recording period in mm (representing Ft0 and Ft180 as depicted in the graph). The fatigue index (FI) is a ratio between Ft180 and Ft0 . FI=Ft180 /Ft0 . View Image -
Figure 5. A typical motor unit's recording to assess functional motor units in normal EDL muscle in mouse is shown. Each trace represents a single‐twitch recording in response to sciatic nerve stimulation. By gradually increasing the stimulating voltage from 0 to 10 V, more motor units are recruited, which contribute incrementally to the amplitude of the recording trace. The number of motor units that innervate the EDL muscle can be calculated by counting the total number of increments (only traces that can be reproduced can be included into the counts). Because of this recording a total of 32 motor units were counted, which are shown as red‐dotted lines. View Image
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Literature Cited
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