Name: repeated measurement of respiratory muscle activity and ventilation in mouse models of neuromuscular disease
Uploaded: 2017-04-17T14:00:00
Description: Watch this Scientific Journal Video about Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease at JoVE.com

Support for this work was provided by a Cincinnati Children&#39;s Hospital Medical Center Trustee Award to S.A.C. and an NIH training grant (T32NS007453) to V.N.J.

Experimental procedures were approved by the Cincinnati Children&#39;s Hospital Medical Center Institutional Animal Care and Use Committee and conducted in compliance with the NIH Guide for the care and use of laboratory animals.
1. Preparing for Telemetry Device Implant Surgery
<ol>
	<li>Put on personal protective equipment (i.e., scrubs, shoe covers, gown, hair net, mask, and surgical gloves). 
	NOTE: This surgery requires a sterile field.</li>
	<li>Turn on the incubator (servo-controlled humidif...

<ol>
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F., Finiels, F., Revah, F., Mallet, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromuscular+function+impairment+is+not+caused+by+motor+neurone+loss+in+FALS+mice:+an+electromyographic+study.">Neuromuscular function impairment is not caused by motor neurone loss in FALS mice: an electromyographic study.</a> Neuroreport. 7 (8), 1427-1431 (1996).</li><li>Pinto, S., Alves, P., Pimentel, B., Swash, M., de Carvalho, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+for+assessment+of+diaphragm+in+ALS.">Ultrasound for assessment of diaphragm in ALS.</a> Clin Neurophysiol. 127 (1), 892-897 (2016).</li><li>Stewart, H., Eisen, A., Road, J., Mezei, M., Weber, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electromyography+of+respiratory+muscles+in+amyotrophic+lateral+sclerosis.">Electromyography of respiratory muscles in amyotrophic lateral sclerosis.</a> J Neurol Sci. 191 (1-2), 67-73 (2001).</li><li>Arnulf, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sleep+disorders+and+diaphragmatic+function+in+patients+with+amyotrophic+lateral+sclerosis.">Sleep disorders and diaphragmatic function in patients with amyotrophic lateral sclerosis.</a> Am J Respir Crit Care Med. 161, 849-856 (2000).</li><li>Bennett, J. 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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+sleep-like+brain+state+alternations+and+breathing+characteristics+in+urethane+anesthetized+mice.">Spontaneous sleep-like brain state alternations and breathing characteristics in urethane anesthetized mice.</a> PLoS One. 8 (7), 70411 (2013).</li><li>Nicaise, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phrenic+motor+neuron+degeneration+compromises+phrenic+axonal+circuitry+and+diaphragm+activity+in+a+unilateral+cervical+contusion+model+of+spinal+cord+injury.">Phrenic motor neuron degeneration compromises phrenic axonal circuitry and diaphragm activity in a unilateral cervical contusion model of spinal cord injury.</a> Exp Neurol. 235 (2), 539-552 (2012).</li><li>Akay, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+measurement+of+muscle+denervation+and+locomotor+behavior+in+individual+wild-type+and+ALS+model+mice.">Long-term measurement of muscle denervation and locomotor behavior in individual wild-type and ALS model mice.</a> J Neurophysiol. 111 (3), 694-703 (2014).</li><li>Tysseling, V. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+potential+early+physiological+marker+for+CNS+oxygen+toxicity:+hyperoxic+hyperpnea+precedes+seizure+in+unanesthetized+rats+breathing+hyperbaric+oxygen.">A potential early physiological marker for CNS oxygen toxicity: hyperoxic hyperpnea precedes seizure in unanesthetized rats breathing hyperbaric oxygen.</a> J Appl Physiol. 114 (1985), 1009-1020 (1985).</li><li>Morrison, J. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+to+remove+ECG+artifacts+from+trunk+muscle+EMG+signals.">A simple method to remove ECG artifacts from trunk muscle EMG signals.</a> J Electromyogr Kinesiol. 19 (6), e554-e555 (2009).</li><li>Lu, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Removing+ECG+noise+from+surface+EMG+signals+using+adaptive+filtering.">Removing ECG noise from surface EMG signals using adaptive filtering.</a> Neurosci Lett. 462 (1), 14-19 (2009).</li></ol>

The procedure demonstrated here allows for the noninvasive (after initial surgical implantation of the transmitter) measurement of respiratory muscle activity and ventilation over many months in the same animal. This technique has several advantages over standard EMG techniques in anesthetized mice: 1) the experiments require fewer mice and provide the ability to record data from the same site in a single mouse across disease stages (instead of using multiple mice at different disease stages); 2) data analysis can be per...

Accessory respiratory muscles (ARMs) increase ventilation during times of high demand (e.g., exercise) and help to maintain ventilation when diaphragm function is compromised following injury or disease1,2. Although changes in diaphragm function have been well described in amyotrophic lateral sclerosis (ALS) patients and mouse models3,4,5,6, much less is known about the activity or function of ARMs in ALS. However, one study suggested that ALS patients that recruit ARMs have a better prognosis than those with similar diaphragm dysfunction that do not7. Furthermore, ARM activity is sufficient for respiration in cases of diaphragm paralysis8. These studies indicate that strategies to augment ARM function may improve breathing in patients suffering from neuromuscular disease, spinal cord injury, or other conditions in which diaphragm function is impaired. However, the mechanisms controlling ARM recruitment for breathing are largely unknown. Methods to measure respiratory function and changes in ARM activity over time in animal models of disease or injury are needed to study how ARMs are recruited, as well as to evaluate therapies to improve ARM recruitment and ventilation. Moreover, the increased activity of ARMs coinciding with the progressive loss of diaphragm function may be a useful biomarker for disease progression in neuromuscular diseases such as ALS7,9,10.
This protocol describes a method to non-invasively (following the initial surgery) and repeatedly measure the activity of respiratory muscles and ventilation in awake, behaving mice. Synchronized recordings of electromyography (EMG), whole-body plethysmography (WBP), and video allow the investigator to assess how changes in ARM activity impact ventilation and to determine when the subject is at rest or moving. A major advantage of this method is that it can be performed in awake, behaving mice, whereas some alternative methods to measure EMG require anesthesia and/or are terminal procedures11,12,13. The recording of EMG activity in awake mice over time may also be accomplished through the chronic implantation of EMG leads, where the mouse is tethered by wires to the acquisition system14,15. Because tethering a mouse could interfere with normal movement or behavior and may not be compatible with a standard plethysmography chamber, the described method uses telemetry devices to wirelessly transmit the EMG signal to the acquisition system. The transmitter can be turned on or off with a magnet to conserve battery power and allows repeated measurements of EMG activity over several months. This protocol can be easily adapted to measure the activity of additional respiratory or non-respiratory muscles by inserting the EMG leads into different muscles. Alternatively, one of the two leads may be used to measure EEG activity to assess sleep state or to identify seizure activity16. This technique has successfully been used to measure changes in ARM activity at rest throughout disease progression in a mouse model of ALS and to identify key neurons driving ARM activity in healthy mice10.

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repeated measurement of respiratory muscle activity and ventilation in mouse models of neuromuscular disease

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated measurements over weeks or months of accessory respiratory muscle activity while simultaneously measuring ventilation in a non-anesthetized, freely behaving mouse. The technique includes the surgical implantation of a radio transmitter and the insertion of electrode leads into the scalene and trapezius muscles to measure the electromyogram activity of these inspiratory muscles. Ventilation is measured by whole-body plethysmography, and animal movement is assessed by video and is synchronized with electromyogram activity. Measurements of muscle activity and ventilation in a mouse model of amyotrophic lateral sclerosis are presented to show how this tool can be used to investigate how respiratory muscle activity changes over time and to assess the impact of muscle activity on ventilation. The described methods can easily be adapted to measure the activity of other muscles or to assess accessory respiratory muscle activity in additional mouse models of disease or injury.

The described protocol was used to implant a telemetry device and to record scalene and trapezius EMG, WBP, and video of a SOD1(G93A) ALS model mouse. Periods in which the animal is inactive (e.g., does not move) were identified using the video recording and confirmed by the lack of movement-related activity in the WBP trace (Figure 3A). Inactive periods include time spent in REM or non-REM sleep, as well as time spent awake but still (Figure 3A)...

Watch this Scientific Journal Video about Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease at JoVE.com

Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease

This paper introduces a method for repeated measurements of ventilation and respiratory muscle activity in a freely behaving amyotrophic lateral sclerosis (ALS) mouse model throughout disease progression with whole-body plethysmography and electromyography via an implanted telemetry device.

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated ...

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Sensitization to grass pollen imposes a global risk for allergic airway diseases. Although prevention relies on local investigation of the pollen ...

Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation and lung parenchymal destruction. It has a very high ...

Assessment of Kidney Function in Mouse Models of Glomerular Disease

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function ...