Name: an innovative running wheel-based mechanism for improved rat training performance
Uploaded: 2016-09-19T13:00:00
Description: Watch this Scientific Journal Video about An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance at JoVE.com

The authors would like to thank Dr. Jhi-Joung Wang, who is the Vice Superintendent of Education at Chi-Mei Medical Center, and Dr. Chih-Chan Lin from the Laboratory Animal Center, Department of Medical Research, Chi-Mei Medical Center, 901 Zhonghua, Yongkang Dist., Tainan City 701, Taiwan, for providing the shooting venue. They would also like to thank Miss Ling-Yu Tang and Mr. Chung-Ham Wang from the Department of Medical Research, Chi-Mei Medical Center, Tainan, Taiwan, for their valuable assistance in demonstrating the prototype system in real experiments with rats. The author gratefully acknowledges the support provided for this study by the Ministry of Science and Technology (MOST 104- 2218-E-167-001-) of Taiwan.

Ethics Statement: The experimental procedures were approved by the animal ethics committee of Southern Taiwan University of Science and Technology Laboratory Animal Center, National Science Council, Republic of China (Tainan, Taiwan).
1. Constructing the Running Wheel Structure
NOTE: All acrylic should be transparent.&#160;Wash the disassembled wheel with water, then use alcohol to wipe the rubber track and acrylic sheets after each use.&#160;&#160;
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This protocol describes a highly effective running wheel system for&#160;reducing the severity of the effects of the stroke&#160;in animals. As a rat-friendly testbed, this platform is designed as well in such a way that a stable running speed can be maintained by rats throughout a running process by means of a predetermined adaptive acceleration curve. In typical training systems, preset training speeds and durations are set manually. Once an exercise commences, a preset speed is reached very shortly. In this context, i...

Strokes exist continuously as a financial burden to countries globally, leaving countless patients physically and mentally disabled1,2. There is clinical evidence to suggest that regular exercise can improve nerve regeneration and strengthen neural connections3,4, and it is also shown that exercise can decrease the risk of suffering ischemic strokes5. With either a treadmill or a running wheel as an exercise training system, rodents, such as rats, serve as a proxy for humans for testing the effectiveness of exercises in a vast majority of clinical experiments6-8. A training system normally involves training a rat for a certain period of time, during which a rat runs at a certain speed. Therefore, the training intensity is generally calculated according to the exercise speed and duration6-8. The same approach is applied to estimate the amount of exercise required for neurophysiological protection. However, the experimental exercises are sometimes found to be ineffective, such as when a rat stumbles, falls, or grabs the rails once they are unable to catch up with the running wheel speed9-11. Needless to say, incidents of ineffective exercise significantly reduce the exercise benefit. Even though there is no any universally accepted approach currently to quantify the effective exercises for reducing brain damage, the level of effective exercises still stands as an objective appraisal for clinical researchers to illustrate the benefits of exercise in the discipline of neurophysiology.
There exist a number of limitations on commercially available animal mobility systems used in today&#39;s brain damage reduction&#160;experiments12. In a treadmill case, rats are forced to run by means of electric shocks, inducing tremendous psychological stress on the animals and thereby interference in the final neurophysiological test results8,13,14. Running wheels can be categorized into two types, namely voluntary and forced. Voluntary running wheels allow rats to run naturally, creating excessive variability due of the differences in the rats&#39; physical traits and abilities15, while motorized running wheels (MRWs) employ a motor to turn the wheel, forcing rats to run. Despite also being a form of forced training, MRWs imposes less psychological stress on rats than treadmills13,16,17. However, experiments using MRWs have reported that rats sometimes interrupt the exercise by grabbing the rails on the wheel track and refusing to run at speeds exceeding 20 m/min9. These examples show that animal mobility systems currently available have an inherent disadvantage that inhibits effective exercising. For objective rat training purposes, the development of a highly effective training system but with low interference is therefore viewed as an urgent issue for neurophysiological exercise experiments.
This study presents a highly effective running wheel system for experiments on&#160;reducing the severity of the effects of the stroke11. In addition to a reduced number of interference factors during a training process, this system detects the running position of a rat using infrared sensors embedded in the wheel, thereby achieving a more reliable estimate of effective exercise activity. The psychological stress imposed by traditional treadmills and the frequent exercise interruptions in MRWs both skew the objectivity of the resulting exercise estimates. A positioning running wheel (PRW) system presented in this study is developed in an attempt to minimize the unwanted interference while providing a reliable training model for quantifying effective exercise.

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			<td height="21" style="height:21px;width:216px;">Brushless DC motor</td>
			<td style="width:195px;">Oriental Motor</td>
			<td style="width:247px;">BLEM512-GFS</td>
			<td style="width:167px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Motor driver</td>
			<td>Oriental Motor</td>
			<td>BLED12A</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Motor reducer</td>
			<td>Oriental Motor</td>
			<td style="width:247px;">GFS5G20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Speedometer</td>
			<td>Oriental Motor</td>
			<td style="width:247px;">OPX-2A</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Treadmill</td>
			<td style="width:195px;">Columbus Instruments</td>
			<td style="width:247px;">Exer-6M</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Infrared transmitter&#160;</td>
			<td style="width:195px;">Seeed Studio</td>
			<td>TSAL6200</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Infrared Receiver</td>
			<td style="width:195px;">Seeed Studio</td>
			<td>TSOP382</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Microcontroller</td>
			<td>Silicon Labs</td>
			<td>C8051F330</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">CCD camera</td>
			<td>Canon Inc.</td>
			<td>EOS 450D</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Image processing software</td>
			<td>Adobe Systems Incorporated</td>
			<td>ADOBE Photoshop CS5 12.0</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Image analysis</td>
			<td>Media Cybernetics</td>
			<td>Pro Plus 4.50.29</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sodium pentobarbital</td>
			<td style="width:195px;">Sigma-Aldrich (Saint Louis, MO, USA)</td>
			<td style="width:247px;">SIGMA P-3761</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ketamine</td>
			<td style="width:195px;">Pfizer (Kent, UK)</td>
			<td style="width:247px;">&#160;1867-66-9</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Atropine</td>
			<td style="width:195px;">Taiwan Biotech Co., Ltd. (Taoyuan, Taiwan)</td>
			<td style="width:247px;">A03BA01</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Xylazine</td>
			<td style="width:195px;">Sigma-Aldrich (Saint Louis, MO, USA)</td>
			<td style="width:247px;">SIGMA X1126</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Buprenorphine</td>
			<td style="width:195px;">Sigma-Aldrich (Saint Louis, MO, USA)</td>
			<td style="width:247px;">B9275</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Anesthesia</td>
			<td>Sigma Chemical</td>
			<td style="width:247px;"></td>
			<td></td>
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an innovative running wheel-based mechanism for improved rat training performance

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity for reducing the severity of the effects of the stroke&#160;in rats. This system provides more effective animal exercise training than commercially available systems such as treadmills and motorized running wheels (MRWs). In contrast to an MRW that can only achieve speeds below 20 m/min, rats are permitted to run at a stable speed of 30 m/min on a more spacious and high-density rubber running track supported by a 15 cm wide acrylic wheel with a diameter of 55 cm in this work. Using a predefined adaptive acceleration curve, the system not only reduces the operator error but also trains the rats to run persistently until a specified intensity is reached. As a way to evaluate the exercise effectiveness, real-time position of a rat is detected by four pairs of infrared sensors deployed on the running wheel. Once an adaptive acceleration curve is initiated using a microcontroller, the data obtained by the infrared sensors are automatically recorded and analyzed in a computer. For comparison purposes, 3 week training is conducted on rats using a treadmill, an MRW and a PRW. After surgically inducing middle cerebral artery occlusion (MCAo), modified neurological severity scores (mNSS) and an inclined plane test were conducted to assess the neurological damages to the rats. PRW is experimentally validated as the most effective among such animal mobility systems. Furthermore, an exercise effectiveness measure, based on rat position analysis, showed that there is a high negative correlation between the effective exercise and the infarct volume, and can be employed to quantify a rat training in any type of brain damage reduction&#160;experiments.

This section is devoted to comparisons, made 1 week after surgery, on the mNSS scores, incline plane test results and brain infarct volumes among five groups. Figure 4A and 4B present the average mNSS scores and the average of incline plane test results, respectively. The PRW group appears as the best in terms of mNSS improvement. The significant differences between PRW and MRW and between treadmill and PRW clearly suggest that the PRW protects against st...

Watch this Scientific Journal Video about An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance at JoVE.com

An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance

This study presents an innovative running wheel-based animal mobility system to quantify an effective exercise activity in rats. A rat-friendly testbed is built, using a predefined adaptive acceleration curve, and a high correlation between the effective exercise rate and the infarct volume suggests the protocol&#39;s potential for stroke prevention experiments.

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity ...

The overall goal of this procedure is to quantify an effective exercise activity for ischemic stroke prevention in rats using a positioning running wheel. This method can help answer key questions in the small medicine field regarding stroke prevention. The main advantage of this technique is that it shows a highly effective running wheel system that can be used for ischemic stroke prevention in rats.The implications of this technique enhance the effectiveness of stroke prevention because the middle cerebral artery encroaching results show our proposed PRW system outperformed the traditional treadmill system. Though this method can provide insight into stroke prevention, it can also be applied to other disease models such as Alzheimer's disease. Demonstrating the procedure will be Ling-Yu Tang and Chung-Ham Wang, technicians from our laboratory.Begin by attaining a transparent acrylic running wheel, 55 centimeters in diameter, and 15 centimeters in width. Cut a quarter circle opening into one side of the running wheel to serve as an entrance and exit for the rats. Then, place a layer of high-friction rubber track on the inside of the acrylic wheel.Next, connect an iron rod with bearings to the running wheel. Then, place two acrylic triangular columns on either side of the running wheel to act as the support frame. Use screws to attach a one millimeter thick semi-circular transparent acrylic sheet to the external sides of the two triangular columns.Ensure that the acrylic sheets are approximately three centimeters away from each side of the running wheel. Use this sheet to position the infrared sensors;to do this, drill a hole in each acrylic sheet at every 45 degree increment in advance. Then, create holes that are the same size as the infrared sensors.To operate the running wheel, use a Brushless DC Motor and a Motor Driver. Mount a rubber disk 10 centimeters in diameter onto the motor's central axis. Using the iron frame and springs to support the motor, connect the rubber disk from the motor's central axis to the outer side of the running wheel.Next, use a microcontroller to rotate the rubber disk, and observe the rotation of the running wheel. Afterwards, mount four infrared emitters on one side, and the corresponding four infrared receivers on the opposite side sequentially between zero degrees and 135 degrees. Finally, connect four pairs of infrared sensors that are mounted in both acrylic sheets to the microcontroller's general pins using single core cables, in order to form the position-running wheel system.Three days before starting the three week training exercise, allow the rat to become familiar with the running environment by manually operating the running wheel. During manually-operated training, gradually accelerate the running speed until the rat is unable to keep pace. Once this occurs, decrease the speed until the rat regains a steady running pace, and then gradually increase the speed again until the rat reaches 20 meters per minute.Three days later, begin the three week exercise training by pressing the start button on the microcontroller each week to execute a training model where the rat runs for 20 meters per minute for 30 minutes during week one, 30 meters per minute for 30 minutes during week two, and 30 meters per minute for 60 minutes during week three. Maintain the rat at a steady state of running between zero degrees and 135 degrees, which is defined as the effective exercise area. Begin by evaluating behavioral performance by neurological severity scores in all stroke rats the day before surgery, as well as daily for seven days after the surgery.Next, measure the limb-strength of the rat using an inclined plane. To do this, place the rat on an inclined climbing apparatus on a daily basis, and acclimatize the rat to the apparatus and the testing conditions one week before testing. During testing, place the rat at the top of the apparatus in a direction where the body axis aligns with the longitudinal axis of the inclined plane.Ensure that the rat stays along the slope of the rubber-ribbed surface of the inclined plane, which should be set at an angle of 25 degrees. Increase the angle dynamically using a ball screw connected to a stepper motor to determine the maximal angle at which the rat can hold onto the plane. Gradually increase the angle of the inclined plane until the rat fails to hold on, and then detect a sliding down event.Finally, have two naive observers independently examine and score all the behavioral tests. This study presented a scientific approach for quantifying effective exercise activity in stroke prevention training, and used neurological severity scores, or M-N-S-S, to verify the proposed method. These results show significant variation among all the exercise and the control groups, indicating that exercise benefits stroke prevention.The PRW group provides the lowest score among the exercise groups, demonstrating a superior neuro-protective mechanism to the other training systems. Additionally, the PRW group acquires a much smaller infarct volume than the control group, and ranks the lowest among all the exercise groups, validating the prominent effect of PRW on stroke prevention. What's this technique can be done in sixteen hours if it is performed properly.While attending this procedure, it's important to remember to rely and follow results and indicator to actually detail the rates, time, and position. Following this procedure, other methods, like a traditional motorized running wheel, can be performed in order to answer additional questions about stable learning. After its development, this technique paved the way for researchers in the field of exercise physiology to explore preventative medicine in rats.After watching this video, you should have a good understanding of how to construct a highly effective running wheel system for ischemic stroke prevention in rats.

Research

JoVE Journal

Behavior

Multifunctional Setup for Studying Human Motor Control Using Transcranial Magnetic Stimulation, Electromyography, Motion Capture, and Virtual Reality

The study of neuromuscular control of movement in humans is accomplished with numerous technologies. Non-invasive methods for investigating neuromuscular ...

Behavioral Disturbances: An Innovative Approach to Monitor the Modulatory Effects of a Nutraceutical Diet

In dogs, diets are often used to modulate behavioral disturbances related to chronic anxiety and stress caused by intense and restless activity. However, ...

Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD

Aerobic exercise (e.g., wheel running (WR) extensively used in animal research) positively impacts many measures of neuroplastic potential in the brain, ...

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific ...

A Vibrotactile Feedback Device for Seated Balance Assessment and Training

Postural perturbations, motion tracking, and sensory feedback are modern techniques used to challenge, assess, and train upright sitting, respectively. ...

Training Dogs for Awake, Unrestrained Functional Magnetic Resonance Imaging

We present a canine functional Magnetic Resonance Imaging (fMRI) training protocol that can be done in a cost-effective manner, with high-energy dogs, for ...

Human Circadian Phenotyping and Diurnal Performance Testing in the Real World

In our continuously developing &#39;around the clock&#39; society, there is a need to increase our understanding of how changes in biology, physiology and ...

Working Memory Training for Older Participants: A Control Group Training Regimen and Initial Intellectual Functioning Assessment

The efficacy of cognitive training interventions is recently highly debated. There is no consensus on what kind of training regimen is the most effective. ...

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila ...