Name: assessing the autonomic and behavioral effects of passive motion in rats using elevator vertical motion and ferris-wheel rotation
Uploaded: 2020-02-07T12:00:00
Description: Watch this Scientific Journal Video about Assessing the Autonomic and Behavioral Effects of Passive Motion in Rats using Elevator Vertical Motion and Ferris-Wheel Rotation at JoVE.com

This work was supported in part by the Hong Kong Research Grants Council, Early Career Scheme, Project #21201217 to C. L. The FWR device has a patent in China: ZL201120231912.1.

The present study and procedures were approved by the Ethics Committee for Animal Experimentation of the Second Military Medical University (Shanghai, China) in accordance with the Guide for the Care and Use of Laboratory Animals (US National Research Council, 1996).
1. Animals
<ol>
	<li>Use Sprague-Dawley (SD) rats of two months (200&#8211;250 g). For each behavioral assay, use a separate group of rats. Always use separate control and experimental groups. 
	NOTE: There were two autonom...

<ol>
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The present study describes assessing autonomic responses to passive motion in rodents using elevator vertical motion and Ferris-wheel rotation. These equipment and procedures can be easily adopted to other rodents and several modifications of the assays exist to confirm vestibular functioning in different circumstances, such as during in pharmacological challenge or surgical interventions. Research in MS elicited by vestibular stimulation has led to the theory that sensory conflict or neuronal mismatch caused by receivi...

Motion sickness (MS) due to abnormal visuo-vestibular stimulation leads to autonomic reaction, eliciting symptoms such epigastric discomfort, nausea and/or vomiting1. According to current theories, motion sickness may be caused by a sensory conflict or neuronal mismatch from receiving integrated motion information that differs from the anticipated internal model of the environment2,3 or postural instability as would occur on a yawing ship4,5. Despite significant advances in the field of motion sickness and vestibular autonomic functioning6,7,8,9,10,11,12, future research can be aided by standardized evaluation protocols. Assessing the autonomic effects of standard passive motions will greatly benefit investigations into the causes and prevention of motion sickness. The overall goal of this study is to assess the autonomic and behavioral effects of passive motion in rodents. Animal models, such as rodents, allow easy experimental manipulation (e.g., passive motion and pharmaceutical) and behavioral evaluation, which can be used to study the etiology of motion sickness. Here, we present a detailed battery for testing the effects of passive motion and the integrity of vestibular functioning.
The present study details two assays, elevator vertical motion (EVM) and Ferris-wheel rotation (FWR), that induce autonomic reactions to the passive motion. The assays are coupled to three quantitative behavioral measures, the balance beam (on mice13 and rats14,15,16,17), open-field examination, and defecation counting. The EVM (similar to the pitch and roll of a ship encountering a wave) assesses vestibular functioning by stimulating the otolith sensory organs that encode linear accelerations (i.e., the saccule that responds to movements in the vertical plane)18. The FWR (centrifugal rotation or sinusoidal motion) device stimulates the otolith organs by linear acceleration and the semicircular canals by angular acceleration19,20. The Ferris-wheel/centrifugal rotation device is unique in its autonomic assessment. To date, the only similar device in the literature is the off-vertical axis rotation (OVAR) turntable, which is used to examine the vestibulo-ocular reflex (VOR)18,21,22, conditioned avoidance23,24, and the effects of hypergravity25,26,27. The EVM assay and the FWR device assay induce vestibular stimulation leading to autonomic reactions. We couple the EVM and FWR to quantitative measurements such as balance beam, defecation counting, and open-field analysis28,29,30, to ensure robust and reproducible results. Similar to those previously described in mice13 and rats14,15,16,17, the balance beam assay is a 1.0 m long beam suspended 0.75 m from the ground between two wooden stools using a simple black-box modification at the goal end (finish). The balance beam has been used to assess anxiety (obscure black box)14,17, traumatic injury15,16,17, and here, autonomic reactions affecting balance. We have performed defecation counting for assessing the autonomic response in the motion sickness model previously, and it is a reliable quantitative measurement that is easily performed and unequivocally assessed6,8,9,11. The open-field analysis employs a simple black box open-&#64257;eld behavior assessment using Ethovision28, Bonsai30, or a simple video analysis in Matlab29 to quantify behavior such as motion. In the current protocol, we use the total distance traveled, but we note several different paradigms exist (e.g., elongation, zone of movement, velocity, etc.)28,29,30. Collectively, these procedures form a short battery of assessments for the examination and evaluation of autonomic reactions to passive motion, for example in motion sickness6,7,8,9,10,11. The present assays can be adapted to a variety of animal models.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Elevator vertical motion device</td>
			<td>Custom</td>
			<td></td>
			<td>Custom-made Elevator vertical motion device to desired specifications</td>
		</tr>
		<tr>
			<td>Ethovision</td>
			<td>Noldus Information Technology</td>
			<td></td>
			<td>Video tracking software</td>
		</tr>
		<tr>
			<td>Ferris-wheel rotation device</td>
			<td>Custom</td>
			<td></td>
			<td>Custom-made Ferris-wheel rotation device to desired specifications</td>
		</tr>
		<tr>
			<td>Latex, polyvinyl or nitrile gloves</td>
			<td>AMMEX</td>
			<td></td>
			<td>Use unpowdered gloves 8-mil</td>
		</tr>
		<tr>
			<td>Open field box</td>
			<td>Custom</td>
			<td></td>
			<td>Darkened plexiglass box with IR camera</td>
		</tr>
		<tr>
			<td>Rat or mouse</td>
			<td>JAX labs</td>
			<td></td>
			<td>Any small rodent</td>
		</tr>
		<tr>
			<td>Small rodent cage</td>
			<td>Tecniplast</td>
			<td>1284L</td>
			<td></td>
		</tr>
		<tr>
			<td>Wooden beam and stools</td>
			<td>Custom</td>
			<td></td>
			<td>Custom-made wooden beam and stools to specifications indicated</td>
		</tr>
	</tbody>
</table>

assessing the autonomic and behavioral effects of passive motion in rats using elevator vertical motion and ferris-wheel rotation

The overall goal of this study is to assess the autonomic and behavioral effects of passive motion in rodents using the elevator vertical motion and Ferris-wheel rotation devices. These assays can help confirm the integrity and normal functioning of the autonomic nervous system. They are coupled to quantitative measures based on defecation counting, open-field examination, and balance beam crossing. The advantages of these assays are their simplicity, reproducibility, and quantitative behavioral measures. The limitations of these assays are that the autonomic reactions could be epiphenomena of non-vestibular disorders and that a functioning vestibular system is required. Examination of disorders such as motion sickness will be greatly aided by the detailed procedures of these assays.

Figure 2 demonstrates representative balance beam results of time taken to transverse. Rats were trained for 3 consecutive days in order to achieve stable performance on the balance beam10. The subsequent day, rats were evaluated for balance beam performance. In the y-axis of the figure, we have the number of seconds taken for rodents to cross the balance beam for Ferris-wheel, elevator vertical motion, and control groups for demonstrative purposes.
<p class="jove...

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Assessing the Autonomic and Behavioral Effects of Passive Motion in Rats using Elevator Vertical Motion and Ferris-Wheel Rotation

Protocols are presented to assess the autonomic and behavioral effects of passive motion in rodents using elevator vertical motion and Ferris-wheel rotation.

The overall goal of this study is to assess the autonomic and behavioral effects of passive motion in rodents using the elevator vertical motion and ...

These protocols are important for establishing the integrity and normal functioning of the autonomic system to assess autonomic nervous system functionality and to reveal vestibular system disorders. A vestibular functioning or motor coordination deficit can signal damage to the autonomic system. These techniques can reveal the integrity of autonomic functioning.Behavioral techniques can be susceptible to variabilities introduced by the experimenter. We recommend the same researcher conduct the techniques throughout the experiment and to treat every rodent identically. For elevator vertical motion analysis, place up to four two-month-old 200 to 250 gram rats in a 22.5 by 26 by 20 centimeter plexiglass box and securely fashion the box.Place the box onto the elevator pad of an elevator vertical motion device and turn on the device at the lowest setting for acclimatization. Next, set the amplitude to 22 centimeters up and 22 centimeters down from neutral and start the device with an initial cycle period at 2, 500 milliseconds for five minutes, 2, 000 milliseconds for five minutes, and 1, 500 milliseconds for five minutes. Then set the device to 1, 000 milliseconds for two hours before slowing the device in reverse using periods of 1, 500 milliseconds for five minutes, 2, 000 milliseconds for five minutes, and 2, 500 milliseconds for five minutes.For a Ferris-wheel rotation procedure, place the plexiglass container on a wooden bench and place the rats into the container with the long axis of their bodies perpendicular to the horizontal rotation rod of the Ferris-wheel. After closing the box securely, place a second set of rats with a similar mass into a second plexiglass container with the long axis of the body perpendicular to the horizontal rotation rod on the second arm of the Ferris-wheel rotation device. After securely closing the box, start the Ferris-wheel rotation in a clockwise direction at 16 degrees per second squared to reach an angular velocity of 120 degrees per second.When the Ferris-wheel has reached peak velocity, begin to decelerate at 48 degrees per second squared to reach zero degrees per second. After a one-second pause, have the container continue to rotate in the counterclockwise direction at 16 degrees per second squared to reach an angular velocity of 120 degrees per second before decelerating at 48 degrees second squared to reach zero degrees per second. Then continue the clockwise-counterclockwise rotation for two hour per session for approximately 720 rotations.For balance beam training, first place two wooden 0.75 meter high stools into the experimental field approximately 110 centimeters apart. Place a plastic black 15 by 15 by eight centimeter box on the finish tool and place a narrow 2.5 by 130 centimeter wooden beam between the two stools leaving a 100 centimeter distance between the stool edges from the start stool to the finish stool. Place a lamp at the start stool and turn on the lamp.Then turn off the room light so that the room is as dark as possible and introduce the first rat to the beam in the lighted corner. If the rat does not cross of its volition, gently prompt the animal to cross the beam. After three consecutive days of training, place the trained rat onto the start stool near the light and immediately start a stopwatch.The rat should cross the balance beam rapidly and enter the black box on the finish stool. Stop the stopwatch as soon as the animal's nose enters the dark box on the finish stool. For defecation counting after motion treatment, transfer the rats from the plexiglass container into individual 40 by 40 by 45 centimeter open field boxes and count the number of feces pellets in the plexiglass box attributed to each rodent.For an open field behavior test, use an infrared video camera to record the animals for three minutes and use the resulting video footage to determine the total distance traveled for each animal. Here, representative balance beam results of the time taken to traverse the balance beam after three consecutive days of training are shown. Elevator vertical motion, changes in the Hertz vertical motion, and Ferris-wheel rotation significantly increased defecation.In addition, elevator vertical motion, changes in the Hertz vertical motion, and Ferris-wheel rotation significantly decreased the open field distance traveled. The balance beam training is critical as the rats must be motivated and confident when crossing the beam. Otherwise, their balance will not be measured during the evaluation period.Following the behavioral procedures, the effects of different drugs of interest on autonomic functioning and cell signaling can be assessed using these vestibular stimulation assays.

Research

JoVE Journal

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