Name: a novel single animal motor function tracking system using simple, readily available software
Uploaded: 2018-08-31T13:30:00
Description: Watch this Scientific Journal Video about A Novel Single Animal Motor Function Tracking System Using Simple, Readily Available Software at JoVE.com

This study was supported by the Merit Review Award # B1495-R (to Jeffrey R. Capadona) and the Presidential Early Career Award for Scientist and Engineers (PECASE) (to Jeffrey R. Capadona) from the United States Department of Veterans Affairs Rehabilitation Research and Development Service. Additionally, this work was supported in part by the Office of the Assistant Secretary of Defense for Health Affairs through the Peer Reviewed Medical Research Program under Award No. W81XWH-15-1-0608. The authors acknowledge SOURCE for its summer research funding support. The contents do not represent the views of the United States Department of Veterans Affairs or the United States Government. The authors would like to thank Hiroyuki Arakawa in the CWRU Rodent Behavior Core for his guidance in designing and testing rodent behavioral protocols. The authors would also like to thank James Drake and Kevin Talbot from the CWRU Department of Mechanical and Aerospace Engineering for their help in designing and manufacturing the rodent ladder test.

All procedures and animal care practices were approved by and performed in accordance with the Louis Stokes Cleveland Department of Veterans Affairs Medical Center Institutional Animal Care and Use Committee. The behavioral testing protocol closely follows previously published work12,13.
1. Behavioral Testing: Filming the Tasks
NOTE: Here, the animals were tested for 8 weeks to detect any chronic behavioral ch...

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The most critical portion of the protocol to ensure a strong analysis is the consistent filming. If the videos are well lit and filmed at the correct position as discussed in the first section of the protocol, the system will be able to do a precise analysis. As with any image-processing problem, the work done in preprocessing will make the post-processing more accurate and simple. As such, making sure that the apparatus and animals are well-lit during the testing and any shadows or other motion in the frame is kept to a...

There are many established methods to assess both functional and behavioral motor and cognitive impairments1,2,3. Some of the more commonly employed methods include testing fine motor function via paw placement, stepping, and limb coordination on a ladder test4, testing gross motor function and stress behavior via the open field grid test5,6, and testing for fear, depression, and despair via the forced swim test7,8 or rotor rod9. However, many of these methods rely on human researchers to &#34;score&#34; the animal or to judge its performance subjectively. The need for a subjective human assessment can slow the generation and analysis of the data, as well as present the opportunity for an intentional or unintentional influence of research bias in the study10. Further, subjective assessment of the data also presents the risk of inaccurate data representation, be it through forgetfulness, poor motivation, improper training, or negligence11.
We have recently reported the use of both an open field grid test and a ladder test in rats implanted with intracortical microelectrodes12,13. Due to the novelty of the findings in those studies, we immediately began employing those and additional functional testing in many ongoing studies in the laboratory. In anticipation of unintentional human-generated variability resulting from an increase in the number of subjective evaluators, and to improve the analysis throughput, we set out to create an automated, computer-assisted program to score behavioral testing, and greatly limit the potential for error.
Here, we report on the development of the BVAS. The BVAS uses computer analysis to score an open field grid test and a ladder test as metrics of gross and fine motor function, respectively. The results can be used to elucidate possible motor function deficits caused by injury or disease, regardless of the injury or disease model. The analysis codes can be adapted to account for changes in behavioral testing equipment or to score various metrics of motor function. Therefore, the BVAS can be implemented in many applications, beyond our intended use or the intended use of those currently employed by other laboratories.
Note that the open field grid and ladder tests require video recording. Therefore, each test will require a video camera [1080 p, minimum 15 frames per second (fps)], a laptop, and a room to store the video data. For both tests, place the camera in a centered position, allowing for the whole apparatus to be seen on the frame. Anchor the camera on a tripod or scaffolding so that it does not move during the testing. Keep the edges of the video frame as close to parallel with the edges of the testing apparatus as possible. Be sure the same personnel complete all testing and the room is well-lit with a temperature-controlled system. Use the same room for all animals throughout the course of the testing, with minimal changes to the room. Cereals or banana chips make good rewards to encourage the animals to complete the behavior tests.

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			<td height="40" style="height:40px;width:216px;">Sprague Dawley rats, male, 201-225g</td>
			<td style="width:197px;">Charles River</td>
			<td style="width:163px;">CD</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Webcam HD Pro c920</td>
			<td style="width:197px;">Logitec</td>
			<td style="width:163px;">960-000764</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Excel</td>
			<td style="width:197px;">Microsoft</td>
			<td style="width:163px;">N/A</td>
			<td style="width:267px;"></td>
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			<td height="40" style="height:40px;width:216px;">Matalb 2017a, Computer Vision System Toolbox</td>
			<td style="width:197px;">Mathworks</td>
			<td style="width:163px;">N/A</td>
			<td style="width:267px;"></td>
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			<td height="40" style="height:40px;width:216px;">Open field grid test</td>
			<td style="width:197px;">Made in-house at Case Western Reserve University</td>
			<td style="width:163px;">N/A</td>
			<td style="width:267px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:216px;">Ladder test</td>
			<td style="width:197px;">Made in-house at Case Western Reserve University</td>
			<td style="width:163px;">N/A</td>
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a novel single animal motor function tracking system using simple, readily available software

We have recently demonstrated that implanting intracortical microelectrodes in the motor corteces of rats&#160;results in immediate and lasting motor deficits. Motor impairments were manually quantified through an open field grid test to measure the gross motor function and through a ladder test to measure the fine motor function. Here, we discuss a technique for the automated quantification of the video-recorded tests using our custom Capadona Behavioral Video Analysis System: Grid and Ladder Test, or BVAS. Leveraging simple and readily available coding software (see the Table of Materials), this program allows for the tracking of a single animal on both the open field grid and the ladder tests. In open field grid tracking, the code thresholds the video for intensity, tracks the position of the rat over the 3 min duration of the grid test, and analyzes the path. It then computes and returns measurements for the total distance traveled, the maximum velocity achieved, the number of left- and right-handed turns, and the total number of grid lines crossed by the rat. In ladder tracking, the code again thresholds the video for intensity, tracks the movement of the rat across the ladder, and returns calculated measurements including the time it took the rat to cross the ladder, the number of paw slips occurring below the plane of the ladder rungs, and the incidence of failures due to stagnation or reversals. We envision that the BVAS developed here can be employed for the analysis of motor function in a variety of applications, including many injury or disease models.

Following the methods presented here, rats completed the open field grid and ladder tests 2x per week. The data were analyzed both by using BVAS and manually with a stop-watch by trained and novice reviewers. The results presented are an average of the raw weekly scores from a single non-implanted control animal over an 8 week study, where week &#34;0&#34; corresponds to the baseline testing. Note there was no testing during week 1 as this was a rest week for the surgery animals. Because ...

Watch this Scientific Journal Video about A Novel Single Animal Motor Function Tracking System Using Simple, Readily Available Software at JoVE.com

A Novel Single Animal Motor Function Tracking System Using Simple, Readily Available Software

The current study aimed to automate the quantification of motor deficits in rats. The initial evaluation model assesses motor loss resulting from an intracortical microelectrode implantation in the motor cortex. We report on the development and use of a tracking algorithm using easily adaptable, simple, and readily available coding software.

We have recently demonstrated that implanting intracortical microelectrodes in the motor corteces of rats&#160;results in immediate and lasting motor ...

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