Name: establishing a device for sleep deprivation in mice
Uploaded: 2023-09-22T14:10:00
Description: Watch this Scientific Journal Video about Establishing a Device for Sleep Deprivation in Mice at JoVE.com

This work was supported by grants from the National Natural Science Foundation of China (82230014, 81930007, 82270342), the Shanghai Outstanding Academic Leaders Program (18XD1402400), the Science and Technology Commission of Shanghai Municipality (22QA1405400, 201409005200, 20YF1426100), Shanghai Pujiang Talent Program (2020PJD030), SHWSRS(2023-62), Shanghai Clinical Research Center for Aging and Medicine (19MC1910500), and Postgraduate Innovation Program of Bengbu Medical College (Byycxz21075).

All animal experimental protocols in this study were approved by the Laboratory Animal Welfare Ethics Committee of Renji Hospital, School of Medicine, Shanghai Jiao Tong University. Male C57BL/6J mice, aged between 8 to 10 weeks, were used in the study. The animals were obtained from a commercial source (see Table of Materials). The major parts required for establishing the device are listed in Figure 1A.
1. Preparation of the sleep deprivat...

<ol>
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Mouse models of sleep deprivation are essential for studying the effects of sleep disruption on various diseases, including cardiovascular disease21, psychiatric conditions22, and neurological disorders23. Among the existing sleep deprivation strategies in mice, physical approaches that involve repetitive short-term interruption of sleep are the most commonly used5,7,<sup class="xr...

Circadian rhythm disruption refers to the desynchronization between the external environment or behavior and the endogenous biological clock. One of the most common causes of circadian rhythm disruption is sleep deprivation1. Sleep deprivation not only negatively affects human health but also significantly increases the risk of many diseases, including cancer2 and cardiovascular diseases3. However, the mechanisms underlying the detrimental effects of sleep deprivation remain largely unknown, and establishing sleep deprivation models is essential to enhance our understanding in this regard.
Various methods for sleep deprivation in mice have been reported, such as the use of water platforms4, gentle handling5, sliding bar chambers6, rotating drums7, and cage agitation protocols5,8,9. Sliding bar chambers automatically sweep bars across the bottom of the cage, forcing the mice to walk over them and stay awake. Cage agitation protocols involve placing cages on laboratory orbital shakers, resulting in efficient sleep disruption. While these methods are automatic and effective, they can be expensive when multiple devices are required to run in parallel, especially for specific study designs that involve a large number of sleep-deprived mice needed for circadian gene profiling. On the other hand, water platforms and gentle handling protocols are cheaper and simpler methods commonly used to induce sleep deprivation. However, the water platform does not allow automatic control of prespecified deprivation-rest cycles10,11, and gentle handling requires continuous vigilance from the researchers to disturb sleep. Additionally, other modalities, like rotating drums, can be confounded by social isolation or stress12.
Inspired by the orbital shaker-based method, we aim to introduce a protocol for establishing a rocker platform-based device for sleep deprivation in mice. This method is cheap, effective, minimally stressful, controllable, and automated. The current protocol allows us to create a rocker platform-based device at a cost approximately ten times cheaper than that of orbital shakers, based on our accessibility. This device effectively disrupted sleep in group-housed mice and induced characteristic changes of sleep deprivation with minimal stress response. It will be especially useful for researchers interested in investigating the effects and underlying mechanisms of sleep deprivation on the pathogenesis of multiple diseases, particularly when the study involves multiple-group sleep deprivation in parallel.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.5 mL microcentrifuge tube</td>
			<td>Axygen</td>
			<td>MCT-150-C-S</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL centrifuge tube</td>
			<td>NEST</td>
			<td>602002</td>
			<td></td>
		</tr>
		<tr>
			<td>Adenosine ELISA kit</td>
			<td>Ruifan technology</td>
			<td>RF8885</td>
			<td></td>
		</tr>
		<tr>
			<td>Animal cage</td>
			<td>ZeYa tech</td>
			<td>MJ2</td>
			<td></td>
		</tr>
		<tr>
			<td>Blood glucose meter</td>
			<td>YuYue</td>
			<td>580</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6J Mice</td>
			<td>JieSiJie Laboratory Animal</td>
			<td>N/A</td>
			<td>Age: 8-10 weeks</td>
		</tr>
		<tr>
			<td>Connecting rod</td>
			<td>ShengXiang Tech</td>
			<td>N/A</td>
			<td>Length:&#160; 20 cm</td>
		</tr>
		<tr>
			<td>Cooling fan</td>
			<td>LiMing</td>
			<td>EFB0805VH</td>
			<td>Supply voltage: 5 V; Power consumption: 1.2 W; Air flow: 26.92 cfm; Dimensions: 40 mm * 40 mm * 56 mm</td>
		</tr>
		<tr>
			<td>Corticosterone ELISA kit</td>
			<td>Elabscience</td>
			<td>E-OSEL-M0001</td>
			<td></td>
		</tr>
		<tr>
			<td>EEG/EMG recording and analysis system</td>
			<td>Pinnacle Technology</td>
			<td>8200-K1-iSE3</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD</td>
			<td>20071302</td>
			<td></td>
		</tr>
		<tr>
			<td>mosquito hemostats</td>
			<td>FST</td>
			<td>13011-12</td>
			<td>Surgical instrument</td>
		</tr>
		<tr>
			<td>Motor and motor mount</td>
			<td>MingYang</td>
			<td>MY36GP-555</td>
			<td>Supply voltage: 24 V dc; Shaft diameter: 8 mm; Maximum output torque: 100 Kgf.cm; Maximum output speed: 10 rpm</td>
		</tr>
		<tr>
			<td>NanoDrop 2000c</td>
			<td>Thermo Scientific</td>
			<td>NanoDrop 2000c</td>
			<td></td>
		</tr>
		<tr>
			<td>Power brick adapter</td>
			<td>MingYang</td>
			<td>QiYe-0243</td>
			<td>Input voltage: 110-220V ac; Output voltage: 24 V dc; Outputcurrent: 2 A; Cable length: 2 m</td>
		</tr>
		<tr>
			<td>qPCR commercial kit</td>
			<td>Vazyme</td>
			<td>Q711-02</td>
			<td></td>
		</tr>
		<tr>
			<td>qPCR measurement equipment</td>
			<td>Roche</td>
			<td>480</td>
			<td></td>
		</tr>
		<tr>
			<td>Rectangle platform attached with a screw-compatible steel cylinder</td>
			<td>Customized</td>
			<td>N/A</td>
			<td>Width: 20 cm; length: 25 cm; length of the cylinder: 30 cm, thickness: 2 mm</td>
		</tr>
		<tr>
			<td>Reverse RNA to cDNA commercial kit</td>
			<td>Vazyme</td>
			<td>R323-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Screw and nut</td>
			<td>Guwanji</td>
			<td>N/A</td>
			<td>Inner diameter: 6 mm, 12 mm</td>
		</tr>
		<tr>
			<td>Screw-compatible steel cylinder</td>
			<td>Customized</td>
			<td>N/A</td>
			<td>Length: 300 mm</td>
		</tr>
		<tr>
			<td>Slotted steel channels</td>
			<td>Customized</td>
			<td>N/A</td>
			<td>Length: 400 mm or 500 mm, thickness: 2 mm</td>
		</tr>
		<tr>
			<td>Time contactor</td>
			<td>LiXiang</td>
			<td>DH48S-S</td>
			<td>Supply voltage: 110-220 V ac; Units measured: hours, minutes, seconds; Contact configuration: DPDT</td>
		</tr>
		<tr>
			<td>TRIzol</td>
			<td>Vazyme</td>
			<td>R401-01</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishing a device for sleep deprivation in mice

Circadian rhythm disruption refers to the desynchronization between the external environment or behavior and the endogenous molecular clock, which significantly impairs health. Sleep deprivation is one of the most common causes of circadian rhythm disruption. Various modalities (e.g., platforms on the water, gentle handling, sliding bar chambers, rotating drums, orbital shakers, etc.) have been reported for inducing sleep deprivation in mice to investigate its effects on health. The current study introduces an alternative method for sleep deprivation in mice. An automated rocker platform-based device was designed that is cost-effective and efficiently disrupts sleep in group-housed mice at adjustable time intervals. This device induces characteristic changes of sleep deprivation with minimal stress response. Consequently, this method may prove useful for investigators interested in studying the effects and underlying mechanisms of sleep deprivation on the pathogenesis of multiple diseases. Moreover, it offers a cost-effective solution, particularly when multiple sleep deprivation devices are required to run in parallel.

The established device for sleep deprivation in mice is shown in Figure 1D. At day 7 after sleep deprivation commencement, electroencephalogram (EEG) and electromyography (EMG) monitoring16 indicated that the device significantly reduced sleep duration and increased wakefulness duration in mice (Figure 2A-D). Meanwhile, the current protocol significantly increased adenosine build-up and mRNA levels of <em...

Watch this Scientific Journal Video about Establishing a Device for Sleep Deprivation in Mice at JoVE.com

Establishing a Device for Sleep Deprivation in Mice

The present protocol outlines a method for setting up a cost-effective rocker platform-based device used for inducing sleep deprivation in mice. This device has proven to be effective in causing disruptions in electroencephalogram (EEG)-evidenced sleep patterns, as well as inducing metabolic and molecular changes associated with sleep deprivation.

Circadian rhythm disruption refers to the desynchronization between the external environment or behavior and the endogenous molecular clock, which ...

The mechanisms underlying the detrimental effects of sleep deprivation remain largely unknown, and establishing sleep deprivation models is essential to enhance our understanding in this regard. This method, they are cheap, effective, minimally stressful, controllable, and automated. The current protocol allows us to create a rocker platform based device at a cost approximately 10 times cheaper than that of orbital shakers based on our accessibility.Begin by securing one end of a 50-centimeter slotted steel channel at the middle of a 40-centimeter slotted steel channel with screws. This will make a T-shaped structure. Position the two T-shaped structures upright 30 centimeters apart.Connect the bottoms of structures with a 30-centimeter screw-compatible steel cylinder using screws. Then place a steel rectangle platform in between the two T-shaped structures. Secure each end of a 30-centimeter screw-compatible steel cylinder with two bearings.Using screws, fix a motor mount on one of the T-shaped structures 25 centimeters down from the top. Now install a motor onto this motor mount with screws. Secure a cooling fan with self-locking bands on the T-shaped structure below the motor.Using screws, fix the bearing end of a connecting rod to a platform corner facing the motor and the other end to the motor shaft. Next, use an electric drill to make two four-millimeter holes at each corner of a plastic container. On the left side of the container, drill two additional four-millimeter holes, along with one lower six-millimeter hole.Lock the plastic container on the rectangle platform using self-locking bands through the drilled corner holes. Secure the water bottle on the left side of the plastic container using self-locking bands through the two four-millimeter holes, with the nozzle going through the six-millimeter hole. Now connect the output electrical wires of the power brick adapter to the two terminals of the motor.Then connect the input wires to the time contactor. Press the rightmost sign buttons on the left and right halves of the time contactor respectively until M appears on the mechanical counters on both sides. Follow this action by pressing the middle sign buttons on both halves of the time contactor until 5M appears on the mechanical counters.Complete this setup by pressing the leftmost sign button on the left half of the time contactor until 15M appears on the left mechanical counter. Next, place the mouse in the cage. Then power the time contactor and the cooling fan.At day seven after sleep deprivation commencement, EEG and EMG monitoring indicated that the sleep deprivation device significantly reduced sleep duration and increased wakefulness duration in mice. Compared to the control, increased levels of successful sleep deprivation markers such as adenosine buildup and Homer1a mRNA levels in the brain were observed. However, no significant changes in serum corticosterone levels were observed.After seven days of sleep deprivation, a significant decrease in mice size, body, and thymus weight was observed. Additionally, glucose tolerance was significantly impaired in mice after sleep deprivation. Sleep deprivation also revealed a significant change in the expression patterns of clock genes in the brain, indicating a disruption of the molecular clock.Balance of the connecting rod and the relative position of the connecting rod between the steel rectangle platform and the motor are the most important things. The researcher can use this technique to investigate the effects and underlying mechanisms of sleep deprivation on various health conditions.

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