Name: automated gait analysis in mice with chronic constriction injury
Uploaded: 2017-10-17T14:00:00
Description: Watch this Scientific Journal Video about Automated Gait Analysis in Mice with Chronic Constriction Injury at JoVE.com

This research was supported by Chungnam National University, Korea Institute of Oriental Medicine (KIOM) and a grant of the Korea Health Technology R&#38;D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health &#38; Welfare, Republic of Korea (grant number: HI15C0007).

All experiments were conducted in accordance with the ethical guidelines of the International Association for the Study of Pain and were approved by the Institutional Animal Care and Use Committee at the Chungnam National University (Daejeon, South Korea).
1. Induction of CCI on Sciatic Nerve
<ol>
	<li>House male ICR mice weighing 20 - 25 g under a 12 h light/dark cycle at controlled room temperature (maintained at 20 - 25 &#176;C) and humidity (40 - 60%), with free access to food an...

<ol>
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P., Bories, C., De Koninck, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+up-down+method+(SUDO)+for+measuring+mechanical+nociception+in+rodents+using+von+Frey+filaments.">A simplified up-down method (SUDO) for measuring mechanical nociception in rodents using von Frey filaments.</a> Mol Pain. 10, 26 (2014).</li><li>Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M., Yaksh, T. 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Currently, measurement of mechanical allodynia by using von Frey filaments is the most widely used method in pain animal models to demonstrate tactile hypersensitivity. As animal models for neuropathic pain have continued to be developed, the methodology of assessment for sensory function has been also improved8,9,10,15. In those reports, it has been suggested these modifications can provide a ...

Pathological changes of nervous system induced by trauma, metabolic dysfunction, inflammation, infection, ischemia, or autoimmune disease occasionally result in neuropathic pain, which is defined as a pain arising as a direct consequence of a lesion or disease affecting the somatosensory system1. Neuropathic pain is usually unbearable and unfortunately, conventional analgesics generally do not produce sufficient pain relief2. A major feature of neuropathic pain contains spontaneous and stimulation-evoked (i.e., allodynia and hyperalgesia) pains. Allodynia is a nociceptive response that occurs to normally non-painful stimuli, such as light touch or warm stimulation. Hyperalgesia indicates an enhanced pain response to noxious mechanical and/or thermal stimuli3. Although these two symptoms both critically impair the patient&#39;s quality of life, mechanical allodynia evoked by gentle tactile stimulation is the most aggravating symptom because soft contact is difficult to avoid in everyday life.
To investigate the underlying mechanism and new analgesics for the treatment of neuropathic pain, the precise measurement of the pain response is essential. Numerous neuropathic pain animal models have developed nociceptive responses on the hind paw area because of its high accessibility4,5,6,7. Thus, most pain response assessments have been performed on the plantar or dorsal surface of the hind paw by applying mechanical stimuli using special instruments, such as von Frey filaments. One of the most frequently used method is the up and down method described by Dixon8 and the later modified versions9,10. However, very skilled, experienced experimenters are required to perform the von Frey test and the results may be subjective.
The automated gait analysis system can investigate neurological and neuromuscular disorders by measuring various parameters of walking in freely moving rodents. In a variety of nerve injury animal models, the degree of nociception and the antinociceptive effect of several treatments can be evaluated without adding a pain stimulation11,12,13,14. This analysis system can detect static and dynamic gait parameters, such as: paw print area (the area of the complete paw print that contacts with the floor), paw intensity (the average intensity of the contacting paw area), stride length (the distance between successive placements of the same paw), stance phase (the duration of ground contact for a single hind paw), step sequence (the order in which the four paws are placed on the floor), swing (the duration of the swing phase), and swing speed (computed from stride length and swing duration and expressed as pixels per second). This paper demonstrates the use an analysis system and provides a brief comparison of data with the von Frey test using chronic constriction injury (CCI) neuropathic mice.

<table>
	<tbody>
		<tr>
			<td>0.9% Saline</td>
			<td>JW Pharmaceutical</td>
			<td>N/A</td>
			<td>Vehicle for drugs</td>
		</tr>
		<tr>
			<td>1ml syringe</td>
			<td>BD Plastipak</td>
			<td>300013</td>
			<td>Injecting device</td>
		</tr>
		<tr>
			<td>2, 2, 2-tribromoethanol (97% purity)</td>
			<td>Sigma</td>
			<td>T48402</td>
			<td>Anesthetic</td>
		</tr>
		<tr>
			<td>2-methyl-2-buthanol (99% purity)</td>
			<td>Sigma</td>
			<td>152463</td>
			<td>Solvent for 2, 2, 2-tribromoethanol</td>
		</tr>
		<tr>
			<td>Catwalk Automated gait analysis system</td>
			<td>Noldus</td>
			<td>N/A</td>
			<td>Automatic analysis software of aniaml gait</td>
		</tr>
		<tr>
			<td>Chromic catgut (4-0 thickness)</td>
			<td>AILEE</td>
			<td>C442</td>
			<td>Ligature to make chronic constriction injury on the sciatic nerve</td>
		</tr>
		<tr>
			<td>Gabapentin</td>
			<td>Sigma</td>
			<td>Y0001280</td>
			<td>Analgeisc, Used as a positive control drug in this study</td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>F.S.T</td>
			<td>11051-10</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Heating Pad</td>
			<td>DAESHIN ELECTRONICS</td>
			<td>M-303AT</td>
			<td>Regulation of body temperature</td>
		</tr>
		<tr>
			<td>ICR Mouse</td>
			<td>Samtaco</td>
			<td>N/A</td>
			<td>Experimental animal</td>
		</tr>
		<tr>
			<td>Mersilk (3-0 thickness)</td>
			<td>ETHICON</td>
			<td>W598H</td>
			<td>Suture material for surgical closure of skin</td>
		</tr>
		<tr>
			<td>Micro-Mosquito</td>
			<td>F.S.T</td>
			<td>13010-12</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Micro-scissors</td>
			<td>F.S.T</td>
			<td>14090-09</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>F.S.T</td>
			<td>12002-12</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Povidone Iodine</td>
			<td>Firson</td>
			<td>N/A</td>
			<td>Disinfectant to prevent infection after surgery</td>
		</tr>
		<tr>
			<td>Scalpel blade</td>
			<td>F.S.T</td>
			<td>10010-00 (#10)</td>
			<td>Surgical instrument to make an incision</td>
		</tr>
		<tr>
			<td>Scalpel handle</td>
			<td>F.S.T</td>
			<td>10003-12 (#3)</td>
			<td>Surgical instrument to make an incision</td>
		</tr>
		<tr>
			<td>Von-Frey filaments</td>
			<td>North Coast</td>
			<td>NC12775-99</td>
			<td>Measurement device to test sensory function for mechanical stimulation</td>
		</tr>
	</tbody>
</table>

automated gait analysis in mice with chronic constriction injury

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some disadvantages such as subjective data and the requirement of a skilled, experienced experimenter. To date, a variety of modifications have improved the von Frey method, but it still has a few limitations. Recent reports have suggested that gait analysis produces more accurate and objective data from the neuropathic animals. This protocol demonstrates how to perform the automated gait analysis to determine the degree of neuropathic pain in mice. After several days of acclimation, the mice were allowed to walk freely on the glass floor to illuminate footprints. Then, quantification of the footprints and gait were performed through video clips with automatic analysis of various walking parameters, such as area of paw print, swing time, angle of paw, etc.
The main purpose of this study is to describe the methodology of automated gait analysis and briefly compare it with data from the classical sensory test using von Frey filament.

We have performed the von Frey test and automated gait analysis in CCI mice until 10 days after CCI surgery. For statistical analysis, repeated measures of two-way analysis of variance (ANOVA) determines the overall effects, and Dunnett&#39;s post-hoc analysis was performed to determine the p-value among the experimental groups.
The results shown in Figure 1 indicate the time course of the ...

Watch this Scientific Journal Video about Automated Gait Analysis in Mice with Chronic Constriction Injury at JoVE.com

Automated Gait Analysis in Mice with Chronic Constriction Injury

The precise assessment of pain response in a neuropathic animal model is critical to investigate the pathophysiology of pain diseases and develop new analgesics. We present a sensitive and objective method to determine the sensory function of the rodent hind paw by an automated gait analysis system.

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some ...

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