This disclosure deals with a system and method for inducing sleep loss in animal test subjects such as rodents (e.g., rats and mice). In particular, disclosed subject matter allows investigators to sleep deprive multiple subjects in parallel with an at least partially automated system.
In various animal research protocols, it is necessary to deprive rats or mice of sleep. Some estimates indicate as many as about 3,000 basic science sleep researchers in the United States alone. A common method which has been used by researchers simply involves the researcher gently handling a test subject to keep it awake. However, it is difficult with such approach to simultaneously sleep deprive plural subjects in a way which provides consistent stimulation over time simultaneously in parallel to plural subjects. Further, it is desirable to not stimulate an animal with novel objects that could induce behavioral plasticity.
As experimenters, even if using a gentle handling approach, it is often difficult to accurately sleep deprive more than 2 animals at one time given that towards the latter end of the protocol (e.g., 5 to 6 hours), animals are quite tired and are difficult to keep awake. An automated system would alleviate such discrepancies and also allow users to sleep deprive multiple animals at the same time, if using multiple chambers.
Summary of the Presently Disclosed Subject Matter
The presently disclosed subject matter deals with a system and method for inducing sleep loss in animal test subjects such as rats or mice. A sleep deprivation chamber uses a DC Servo motor with a controller to cause a bar to sweep across the rodent cage floor. The system is programmed to sweep the bar across the floor of the rodent cage at varying intervals. The maximum speed at which the bar sweeps the entire rodent cage may be a set time, for example, such as within 3 seconds. The chamber is designed such that it can structurally accommodate a receiver and can record EEG/EMG telemetrically in animals that have been implanted with sleep transmitters. During the sleep deprivation protocol, food and water are available to animals in the chamber at all times. The chamber is also designed to fit a standardized rat cage, and plural cages, such as an 8-chamber sleep restriction device, can be simultaneously operated.
The presently disclosed animal protocol allows us to use plural chambers to deprive a plurality of rats of sleep. Some advantages of such chambers over, for example, a currently used gentle handling method:
In one exemplary embodiment disclosed herewith, methodology for inducing sleep loss in animal test subjects may preferably comprise providing an animal enclosure cage with a removable lid forming an elongated slot along the top of the enclosure cage; mounting a controllably movable arm adjacent to the cage; positioning the movable arm for projection into the cage through the elongated slot so that at least a portion of the arm is movable adjacent to the floor of the cage; and controllably moving the movable arm along the cage floor so that an animal test subject in the cage is periodically moved to deprive sleep of the animal test subject.
It is to be understood that the presently disclosed subject matter equally relates to associated and/or corresponding systems.
Other example aspects of the present disclosure are directed to systems, apparatus, tangible, non-transitory computer-readable media, user interfaces, memory devices, and electronic devices related to sleep deprivation. To implement methodology and technology herewith, one or more processors may be provided, programmed to perform the steps and functions as called for by the presently disclosed subject matter, as will be understood by those of ordinary skill in the art.
Another exemplary embodiment of presently disclosed subject matter relates to a system for inducing sleep loss in animal test subjects, preferably comprising a removable lid configured to fit on a preexisting animal enclosure cage, with an elongated slot formed along the top of the enclosure cage; a guide rail situated adjacent to the cage; a movable arm received on the guide rail for movement there along; and a controllable motor for controllably driving the arm relative to the guide rail. Per such exemplary system, preferably the movable arm is an L-shaped arm having at least a portion thereof which is positioned to extend through the elongated slot and along the width of the cage floor while the arm is received on the guide rail, so that controllably moving the arm through operation of the controllable motor moves the movable arm along the cage floor so that an animal test subject in the cage is periodically moved to deprive sleep of the animal test subject.
Yet another exemplary embodiment of presently disclosed subject matter relates to an automated system for inducing sleep loss in rat test subjects according to a programmed sleep deprivation protocol. Preferably, such exemplary automated system may comprise a rat enclosure cage; a removable lid configured to fit on the cage, with an elongated slot formed along the top of the cage; a guide rail supported adjacent to the cage; a movable sweep arm received on the guide rail for movement back and forth along the rail; a controllable motor for controllably driving the sweep arm relative to the guide rail; and a programmable motor controller for controlling operation of the motor according to a programmed sleep deprivation protocol. Further, preferably, such sweep arm is L-shaped with a base portion thereof positioned to extend through the elongated slot and along the width of the cage floor while the arm is received on the guide rail, so that controllably moving the arm through operation of the controllable motor moves the movable arm along the cage floor so that a rat test subject in the cage is periodically moved to deprive sleep of the rat test subject according to the programmed sleep deprivation protocol.
Additional objects and advantages of the presently disclosed subject matter are set forth in, or will be apparent to, those of ordinary skill in the art from the detailed description herein. Also, it should be further appreciated that modifications and variations to the specifically illustrated, referred and discussed features, elements, and steps hereof may be practiced in various embodiments, uses, and practices of the presently disclosed subject matter without departing from the spirit and scope of the subject matter. Variations may include, but are not limited to, substitution of equivalent means, features, or steps for those illustrated, referenced, or discussed, and the functional, operational, or positional reversal of various parts, features, steps, or the like.
Still further, it is to be understood that different embodiments, as well as different presently preferred embodiments, of the presently disclosed subject matter may include various combinations or configurations of presently disclosed features, steps, or elements, or their equivalents (including combinations of features, parts, or steps or configurations thereof not expressly shown in the figures or stated in the detailed description of such figures). Additional embodiments of the presently disclosed subject matter, not necessarily expressed in the summary section, may include and incorporate various combinations of aspects of features, components, or steps referenced in the summarized objects above, and/or other features, components, or steps as otherwise discussed in this application. Those of ordinary skill in the art will better appreciate the features and aspects of such embodiments, and others, upon review of the remainder of the specification, will appreciate that the presently disclosed subject matter applies equally to corresponding methodologies as associated with practice of any of the present exemplary devices and vice versa.
These and other features, aspects and advantages of various embodiments will become better understood with reference to the following description and appended claims. The accompanying figures, which are incorporated in and constitute a part of this specification, illustrate embodiments of the present disclosure and, together with the description, serve to explain the related principles.
A full and enabling disclosure of the present subject matter, including the best mode thereof to one of ordinary skill in the art, is set forth more particularly in the remainder of the specification, including reference to the accompanying figures in which:
Repeat use of reference characters in the present specification and figures is intended to represent the same or analogous features or elements of the present invention.
Reference will now be made in detail to various embodiments of the disclosed subject matter, one or more examples of which are set forth below. Each embodiment is provided by way of explanation of the subject matter, not limitation thereof. In fact, it will be apparent to those skilled in the art that various modifications and variations may be made in the present disclosure without departing from the scope or spirit of the subject matter. For instance, features illustrated or described as part of one embodiment may be used in another embodiment to yield a still further embodiment.
In general, the present disclosure is directed to a system and method for inducing sleep loss in animal test subjects, such as rats. A sleep deprivation chamber uses a controlled motor to cause a bar to sweep across the rodent cage floor at varying intervals. The chamber or enclosure is designed such that it can structurally accommodate a receiver and can record EEG/EMG telemetrically in animals that have been implanted with sleep transmitters. Food and water remain fully available to animals at all times in the chamber. In some embodiments, a presently disclosed chamber may be also designed to fit a standardized rat cage. In other embodiments, plural cages such as an 8-chamber sleep restriction device can be simultaneously operated.
To induce sleep loss in rats, we have designed a sleep deprivation chamber using for example a Teknic™ 24v DC Servo motor (Teknic, Inc., Victor, NY) that operates with for example a Velocio PLC (Velocio Networks, Huntsville, Alabama) to sweep a bar across the rodent cage floor. The programmable logic controller (PLC) may be programmed for example with Velocio vBuilder software, an open source coding program, and use, for example, a Velocio monitor that is connected via HDMI. The system may be programmed to sweep the bar across the floor of the rodent cage at varying intervals. The maximum speed at which the bar sweeps the entire rodent cage may be set, for example, such as within 3 seconds. The chamber is designed such that we can place it over a Data Science International (DSI; a division of Harvard Bioscience, Inc.; Holliston, MA) receiver and record EEG/EMG telemetrically in animals that have been implanted with sleep transmitters. During the sleep deprivation protocol (e.g., 6 hours), food and water are available to animals in the chamber at all times. The chamber is also designed to fit a standard rat cage from veterinary resources and cages can be replaced upon completion of an experiment.
Plural subjects may be addressed in parallel. For example, we have developed one exemplary embodiment which comprises an 8-chamber sleep disturbance apparatus. It is functional for use with rodents as shown by feasibility testing. For example, such 8-chamber sleep restriction chamber can be used by neuroscientists to prevent small animals, such as rats and mice, from initiating into sleep. The chamber works in an automated fashion so that it sweeps a bar across the floor of the rodent cage. This will physically nudge the animal and prevent it from falling asleep. Thus, it allows, in automated fashion, to sleep restrict and sleep deprive a plurality of small animals and test hypotheses related to the role of sleep.
The presently disclosed chambers are designed to work with a basic sleep monitoring system that many neuroscientists use (e.g., the DSI telemetric EEG/EMG system). The chambers can also be programmed to work simultaneously and increase throughput. As compared with stand-alone models, the multi-chamber models disclosed herewith increase efficiency, improve accuracy, and increase productivity. In addition, the presently disclosed technology facilitates incorporation of Artificial Intelligence routines to autonomously operate without the need for constant human intervention.
One presently disclosed exemplary control protocol for sleep deprivation of subjects is represented by
In particular, another Sleep Deprivation Chamber Protocol may be represented as follows:
For another exemplary protocol arrangement, one can incorporate the use of the sleep deprivation chamber (
The following disclosure indicates in effect a “parts” list for such an exemplary embodiment as the associated Monitor and other features of the presently disclosed subject matter.
Further, the outline (near right top) in
Representing further exemplary features,
The following discloses exemplary operation of an exemplary embodiment of the presently disclosed subject matter.
When power is first turned on to the system, the motors will start flashing orange in the back, as represented by
Section 2-2 Entering a Cage into the System
When the motors are in the home position, the user will remove the Metal Door from the Plastic Cage Cover, as shown progressively by
One of the options in programing is to determine how many motors are allowed to run during an experiment. This option can be found in the top righthand corner of the monitor, as shown in
The Delay function, as referenced by
For a user to check whether the program has started, the Gold Page Button in the bottom righthand corner of the monitor screen (see
Step 1: A user may enter in the Total Run Time in the boxes in the top center of the Monitor screen. Time can be entered in as Hours and Minutes which will combine to create the total run time for the program, as represented in
Step 2: A user may select how many segments are needed by pressing the Duration Selection Buttons, as shown in
Step 3: A user selects what Time Value is needed for the Duration Time. This can be either Hours or Minutes. The default value for each Duration is in Hours but can be changed to Minutes by pressing the Duration Time Value Selection Button shown in
Step 4: A user enters in the amount of time the Duration will last for into the Duration Length Entry Location, as represented by
Step 5: The user selects what Time Value is needed for the Pause Time. The default values for a Pause is set in Minutes. The value can be changed by pressing on the Pause Time Value Selection Button shown in
Step 6: A user may enter in the amount of time the Pause will last into the Pause Length Entry Location, as represented by
Step 7: Steps 3 through 6 are repeated for each Duration needed for the program. When the program is fully entered, the user proceeds to the next section.
After checking all of the Values and the program is ready to Start, the user presses the Start Button. After pressing the Start Button, the user checks either the Program Running Timer or the Delay Timer if a delay is set. If either Timer is counting up, it signifies that the program has started, as referenced in
If at any point the program must be stopped, the user presses the Stop Button. After pressing the Stop Button, the user checks the Program Running Timer. If it has stopped counting, the user has successfully stopped the program. If the Stop Button is pressed in the middle of a motor movement, the motor will continue to move until it reaches either Point A or Point B, as illustrated in
If at any point a Program needs to be Reset to the beginning, the program must be Stopped first. Once the program is Stopped, the user presses the Reset Button to Reset the Program. To confirm that the Program has been Reset, the user checks the Program Running Timer. If it shows 0s across the board, the Program has been Reset.
When a Program finishes, it will clear out the Program Running Timer but will keep all other values. If a Program needs to be run again, the user may do so by pressing the Start Button. The user should keep in mind that the Delay option turns itself off after it has finished its operation. Thus, in order to Delay the start of the Program, it must be turned back on, and the Delay Time reentered again. When the user is done with experiments and the system is no longer needed, the user should turn the power off. This can be done by either unplugging everything or by turning a power strip off if the Chamber is plugged into it. This is done to allow the system to rest and to remove all stress on the motors waiting for commands.
While certain embodiments of the disclosed subject matter have been described using specific terms, such description is for illustrative purposes only, and it is to be understood that changes and variations may be made without departing from the spirit or scope of the subject matter. For example, various and different times and time periods may be used in a given protocol, as compared to the exemplary times and time periods disclosed herewith.
The present application claims the benefit of priority of U.S. Provisional Patent Application No. 63/390,883, titled Sleep Disturbance Chamber For Animal Test Subjects, filed Jul. 20, 2022, and which is fully incorporated herein by reference for all purposes.
This invention was made with government support under Grant No. RO1 NS102209, awarded by National Institute of Neurological Disorders and Stroke (NINDS), an Institute of the National Institutes of Health, and Grant No. 2P20GM103499-20, awarded by the National Institute of General Medical Sciences (NIGMS), an Institute of the National Institutes of Health (NIH). The government has certain rights in the invention.
Number | Date | Country | |
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63390883 | Jul 2022 | US |