WO2023043290A1

WO2023043290A1 - Automated system and method for training or measuring multiple cognitive behaviors of laboratory animals

Info

Publication number: WO2023043290A1
Application number: PCT/KR2022/013965
Authority: WO
Inventors: 이승희
Original assignee: 한국과학기술원
Priority date: 2021-09-17
Filing date: 2022-09-19
Publication date: 2023-03-23
Also published as: KR102344513B1

Abstract

The present invention pertains to: an automated system for training or measuring multiple cognitive behaviors, wherein laboratory animals can automatically receive behavior training or be measured without human intervention by inducing the laboratory animals to enter a behavior experiment space on their own after staying freely inside a cage; and a method for using the automated system to train the cognitive ability of the brain with respect to multi-sensory information, and accurately and quantitatively measuring changes in the cognitive ability of the brain. In addition, the present invention pertains to a drug screening method for using the automated system according to the present invention to verify the effect that targeted drugs being developed to prevent or treat brain cognitive dysfunction or loss have on changes in brain function, or a method for using the automated system according to the present invention to verify the effectiveness of digital therapeutics using various multi-sensory stimuli.

Description

Automated system and method for training or measuring multiple cognitive behaviors of laboratory animals

The present invention is an automated system that allows laboratory animals to automatically perform multi-cognitive behavioral training or measurement without human intervention by inducing them to enter the behavioral experiment space by themselves after freely staying in a life cage, and brain development using the same. It relates to a method for quantitatively measuring changes in cognitive ability. In addition, the present invention is a drug screening method or various multi-sensory drug screening method for verifying the effect of a target drug developed for the prevention or treatment of brain cognitive dysfunction or loss using the multi-cognitive behavioral training or measurement automation system on changes in brain function It relates to a method for verifying the effectiveness of digital therapeutics using stimulation.

This result was studied with the support of the Personal Basic Research Project of the Ministry of Science and ICT (2021R1A2C3012159).

In order to prevent or treat impairment or loss of brain cognitive function, it is necessary to identify neural networks that play an important role in brain cognitive ability in vivo. To this end, it is necessary to quantitatively measure the subtle changes in cognitive abilities of the brain in situations where various external stimuli are simultaneously applied.

In general, experiments that measure changes in brain cognitive abilities in response to external sensory stimuli, such as hearing, sight, touch, or smell, require training of the subject prior to measurement. In addition, since the experimenter should be able to judge whether the trained stimulus is properly recognized in the brain, studies have been conducted mainly on higher intelligent animals such as humans or monkeys.

However, in order to develop a drug or treatment method that treats brain function, it is necessary to check the real-time gene expression regulation pattern that changes after administration of a target drug, or to determine whether or not cognitive ability has been restored or changed in a brain that has actually lost function through action. Experiments with higher animals such as humans or monkeys have many limitations because various approaches are required, such as the process of examining and histologically confirming any changes.

Accordingly, studies on brain function are being actively conducted using mice or rats, which are small animals among mammals having brain structures similar to those of humans. However, research and development is very difficult because a method and measurement system for quantitatively measuring brain perceptible changes in multi-sensory information of small animals such as mice have not been developed.

In addition, when a mouse or rat is used as an experimental animal, it is preferable to conduct a behavioral experiment at night time in consideration of the physiological characteristics of rodents active at night. In addition, behavioral experiments can only derive meaningful results that can link behavioral changes and histological changes only by analyzing large amounts of data obtained through large-scale repeated experiments, so considerable time and manpower are consumed to obtain behavioral experiment results.

In order to solve this problem, an automated behavior training system recognizing multi-sensory information that allows the experimental animals to freely enter the behavioral experiment space after staying in the life cage so that the experimental animals can learn and train themselves without human intervention need this

Looking at the prior art, in order to observe the behavior of the animal, a mouse is confined to a cage of a certain size, and an animal behavior experiment apparatus for recording and analyzing the movement of the mouse using imaging equipment such as an infrared camera installed at the top of the cage is disclosed. There is a bar (Korean Registered Patent Publication No. 10-1929570, 2018. 12. 14. Announcement)

However, the animal behavior experiment apparatus, which analyzes recorded images later to determine behavior, is very difficult to quantitatively measure minute changes in brain cognitive ability in experimental animals responding to multiple sensory stimuli that simultaneously or selectively stimulate vision and hearing. It is difficult. In addition, there is a high possibility that the user will intervene in the experiment and affect the behavioral experiment, and it is very difficult to obtain a large amount of behavioral experiment measurement data.

As another prior art, an apparatus and method for automating animal odor discrimination training have recently been disclosed (Korean Patent Publication No. 10-2021-0087302, published on July 12, 2021). In detail, the present invention relates to an apparatus for automating smell discrimination training of animals and a method thereof, which can increase the utilization of olfactory-based animals by shortening the odor discrimination training period and reducing costs. When using this device, the control unit automatically controls the operation of each device according to a pre-entered program so that the animal can be trained.

However, in order to repeatedly conduct experiments to measure changes in the brain cognitive ability of laboratory animals, it is necessary to provide a system for training the animals themselves, and to collect large-scale data quantitatively from this, and to fix the body of the laboratory animals Experiments to observe changes in brain function are also required, but it is very difficult to implement the above experimental conditions with the prior art.

For this reason, in the prior art animal training system, brain cognitive impairment or loss is mild cognitive impairment, vascular cognitive impairment, cognitive impairment due to cerebral hemorrhage, dementia, brain damage, memory impairment, visual and/or auditory discrimination impairment, autism, attention It is difficult to use as a drug screening device and method for the treatment and prevention of attention deficit hyperactivity disorder (ADHD), schizophrenia or Parkinson's disease.

In addition, a large-scale animal behavior experiment apparatus is required to verify the effect of digital therapeutics, which are programs for treating brain cognitive impairment or loss, which are being actively developed recently. However, it is very difficult to obtain a large amount of behavioral experiment data for the development of a treatment method for digital therapeutics or the verification of treatment effects with conventional laboratory animal devices and methods, making it difficult to utilize them.

Accordingly, there is an urgent need for an automated behavioral training system for measuring multiple cognitive ability changes in the brain and a method for using the same, which allows laboratory animals to perform learning training according to multiple sensory information stimuli without human intervention.

[Prior art literature]

[Patent Literature]

(Patent Document 1) Republic of Korea Patent Registration No. 10-1929570

(Patent Document 2) Republic of Korea Patent Publication No. 10-2021-0087302

The present invention was invented to solve the above conventional problems, and by inducing the experimental animals to enter the behavioral experiment space by themselves by quenching thirst due to water shortage or craving for food after staying freely in the life cage, An object of the present invention is to provide an automated system for multi-cognitive behavioral training or measurement that enables laboratory animals to perform learning behavioral training by themselves without intervention, and a method of using the same.

Changes in multi-sensory information cognitive ability of the brain can be quantitatively measured using the experimental animal model learned according to the apparatus or method of the present invention.

In addition, the present invention aims to be used in non-clinical tests to verify the effect of a target drug developed for the prevention or treatment of brain cognitive dysfunction or loss on changes in brain function.

Furthermore, the multi-cognitive behavioral training automation system and method of the present invention enable simultaneous behavioral experiments of large-scale experimental animals and acquire large-scale behavioral test data by utilizing it, so development and development of digital therapeutics that can treat damaged brain functions and It is intended to be used for efficacy verification.

Other objects of the present application and technical problems to be achieved by the present invention are not limited to the above-mentioned objects, and will become more clear by the following description of the invention together with the claims and drawings described in this specification.

In addition, the technical problem to be achieved by the present invention, which is not described in the present specification, will be clearly understood and inferred by those skilled in the art (hereinafter referred to as 'common technology').

In order to achieve the above object, an automated system for training or measuring multiple cognitive behaviors provided by the present invention and a method of using the same are as follows.

An automated system for multi-cognitive behavioral training or measurement in which a laboratory animal can perform behavioral training by itself according to the present invention includes a head plate 10 mounted on the head of the laboratory animal; a behavior experiment unit 120 in which cognitive behavioral training or experiments of laboratory animals are performed; A head fixing unit 20 located within the behavior experiment unit and coupled to the head plate 10 to fix the lab animal; the behavior experiment unit 120 includes a space where the lab animal normally lives and Is a separated space or a space in which laboratory animals can freely move and enter, and the head plate 10 is automatically bound to the head fixing part 20 through mechanical, magnetic or electromagnetic means. can

In addition, the automated system may include an experimental animal having the head plate 10 mounted on its head; A housing 100 formed of an opaque panel on an entire surface to block external light; a life cage (130) located within the housing and in which the laboratory animals freely live; And an actuator 30 located in the behavior experiment unit 120 and connected to the head fixing unit 20 to move up and down; Including, the actuator may further include a stepping motor, a rack and a pinion gear. Here, the head plate may be surgically implanted into the head of an experimental animal.

In addition, the automated system for training or measuring multiple cognitive behaviors according to the present invention includes a stimulus providing unit 140 that is located on the front or side of the behavior experiment unit 120 and applies selected sensory stimuli to the experimental animals; a behavior detection unit 110 for applying a reward or punishment according to the cognitive behavior of the laboratory animal; a water supply unit 180 that is connected to the behavior experiment unit 120 and controls water supply to lab animals; a data acquisition unit 150 that automatically measures a water intake behavior according to the recognition of sensory information of the laboratory animal through the signal detected by the behavior detection unit 120; a power supply unit 170 supplying electricity to circuit devices included in the housing 100; Training software (not shown) for measuring and training the cognitive abilities of the laboratory animals; A microcomputer 160 controlling the operation of the device within the housing 100 and storing recorded data may be further included.

In one embodiment, the life cage 130 includes a connection unit 125 connected to the behavior experiment unit 120; The connection part 125 may further include a touch sensor for detecting the movement of the test animal and an automatic shutter 40 for allowing or restricting the movement of the test animal according to the detection of the touch sensor.

In one embodiment, the head plate 10 may include at least one permanent magnet or metal plate, and the head fixing part 20 may include at least one permanent magnet or electromagnet bound to the head plate 10. And, the actuator 30 connected to the head fixing part 20 and moving up and down may include a stepping motor, rack and pinion gears (not shown).

In one embodiment, a water bottle 310 attached to the outside of the housing 100 and supplying water to the water supply unit 180, a water bottle mounting unit 320 for fixing the water bottle to the housing, and a connection for supplying water. A tube 330 may be further included, and the water supply unit may further include one or more water compensation ports 50 and a mini pump 182 for controlling the operation of each water compensation port.

In one embodiment, a touch screen 300 capable of inputting control commands to the upper surface of the housing, confirming test results, and monitoring the condition of the test animal inside the housing in real time may be further included.

In one embodiment, the stimulus providing unit 140 may include at least one speaker 141 for applying auditory stimuli to the lab animal, and at least one monitor 142 for applying visual stimuli.

In one embodiment, the action detection unit 110 includes a water compensation port 50 in which water comes out for a certain period of time when the learned action is successfully recognized, and air comes out as a punishment instead of water when it fails to recognize the learned action. An air puff 60 may be included.

In one embodiment, the housing may include a lighting unit 190 installed therein to provide light so that the laboratory animal distinguishes between day and night, and a timer 191 capable of setting an operating time of the lighting unit.

In one embodiment, the data acquisition unit 150 measures the time and number of times when a behavior of the lab animal licking the water compensation port occurs in the behavior detection unit and converts it into digital data. Includes a Data Acquisition system (DAQ) can do.

In one embodiment, the power supply unit 170 may include at least one power output terminal 171 supplying DC power of 5V to 24V to the inside of the housing.

In one embodiment, an infrared camera 70 located on one side of the upper portion of the behavior experiment unit 120 may further include an infrared camera 70 for monitoring the state of the laboratory animal even in a dark state without light inside the housing 100 .

In one embodiment, the laboratory animal may be an laboratory animal having a brain cognitive function disorder or loss.

In one embodiment, the automation system can be stacked using a rack 220 to train a plurality of laboratory animals at the same time by connecting at least one or to obtain a large amount of cognitive ability measurement data, , can be connected to the data server 200 by wire or wirelessly.

In addition, the automation method for training or measuring multiple cognitive behaviors of laboratory animals using the automated system for training or measuring multiple cognitive behaviors according to the present invention is: Moving to the behavior experiment unit 120 by itself; (b) automatically binding the head fixing part 20 to the head plate 10 of the experimental animal when it is detected that the experimental animal entered the behavioral testing unit 120; (c) separating the head fixing part from the head plate when the training or experiment is finished according to the set training program; (d) returning the experimental animal to the life cage outside the behavioral experiment unit; characterized in that it comprises a.

In one embodiment, the experimental animal may be an experimental animal model with brain cognitive dysfunction or loss.

In one embodiment, the brain cognitive impairment or loss is mild cognitive impairment, vascular cognitive impairment, cognitive impairment due to cerebral hemorrhage, dementia, brain damage, memory impairment, visual and/or auditory discrimination impairment, autism, attention deficit hyperactivity disorder (Attention deficit hyperactivity disorder, ADHD), schizophrenia or Parkinson's disease.

In one embodiment, in the automated method for training or measuring multiple cognitive behaviors, after the step (d), the effect verification step (e step) may be further included.

The present invention uses an automated system for training or measuring multiple cognitive behaviors and measures brain cognitive dysfunction including the step of measuring changes in brain cognitive ability in experimental animals administered with a drug candidate exhibiting preventive or therapeutic effects on brain cognitive function. Or it provides a method for screening drug candidates exhibiting preventive or therapeutic effects on damage.

In one embodiment, the experimental animal may have mild cognitive impairment, vascular cognitive impairment, cognitive impairment due to cerebral hemorrhage, dementia, brain damage, memory impairment, visual and/or auditory discrimination impairment, autism, attention deficit hyperactivity disorder (Attention deficit hyperactivity disorder) disorder, ADHD), schizophrenia or Parkinson's disease.

The present invention also provides a method including a step of verifying the effect of a digital therapeutic program, which is a program for alleviating or treating symptoms of brain cognitive dysfunction or loss, by using an automated system for training or measuring multiple cognitive behaviors.

An automated system for training or measuring multiple cognitive behaviors according to the present invention and a method using the same can obtain a large amount of behavioral experiment data in a short period of time because large-scale laboratory animals can simultaneously perform learning behavioral training on their own without human intervention.

Using the experimental animal model learned using the automated system and method of the present invention, it is possible to more accurately and quantitatively measure the change in multi-sensory information cognitive ability of the brain.

According to the present invention, it is easy to precisely control sensory stimuli applied from the outside, and thus it is possible to measure the effect of various drugs and quantitatively measure the actual brain activity state in real time.

In addition, the present invention can be used in a non-clinical test to verify the effect of a target drug developed for the prevention or treatment of brain cognitive dysfunction or loss on changes in brain function, and can be used for research and development to screen drug candidates. It can also be used to develop digital therapeutics to treat or prevent brain cognitive dysfunction or loss.

1 is a view of a head plate 10 surgically fixed to the head of a laboratory animal according to an embodiment of the present invention.

2 is a schematic diagram of an automated system for training or measuring multiple cognitive behaviors according to an embodiment of the present invention.

3 is an internal view of the housing 100 of an automated system for training or measuring multiple cognitive behaviors according to an embodiment of the present invention.

4 is a view of a water bottle 310, a connection tube 330, and a touch screen 300 mounted outside the housing 100 according to an embodiment of the present invention.

5 is an example of the appearance of the automatic shutter 40 connected to the life cage 130 according to an embodiment of the present invention.

6 is an example of how the head fixing part 20 according to an embodiment of the present invention is fixed to the head plate 10 of a lab animal.

7 is an example of a water supply port 50 located in front of an experimental animal whose head is fixed according to an embodiment of the present invention.

8 is an example of an internal infrared camera 70 for monitoring mice in training according to an embodiment of the present invention.

9 is an example of a data server 200 and a rack 220 that acquire a large amount of data by simultaneously connecting one or more automated systems for training or measuring multiple cognitive behaviors according to an embodiment of the present invention.

10 is an example showing results of multi-cognitive behavioral training of laboratory animals according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail so that a person skilled in the art can easily implement the present invention through examples and accompanying drawings.

It is obvious that the content of the invention disclosed by this specification is not limited to the content of the specific embodiments described herein and the accompanying drawings, and includes that a person skilled in the art can implement it in various ways in the technical field to which the present invention belongs. do.

Therefore, it should be understood that the content of the invention disclosed in this specification is not limited to the specific embodiments described herein, and modifications and other embodiments thereof are also included within the scope of the claims.

Referring to FIG. 1, the head plate 10 includes at least one permanent magnet or metal plate, and the head fixing part 20 of the actuator 30 coupled to the head plate includes at least one permanent magnet coupled to the head plate. Magnets or electromagnets may be included.

The head plate may be made of a non-magnetic metal that does not adhere to magnets, such as titanium, tungsten, or aluminum. Alternatively, it may be made of any one material selected from polyether ether ketone (PEEK), thermoplastic elastomer (TPE), and polyphenylsulphone (PPSU), which are widely used as medical plastics.

When one or more

permanent magnets

11 and 12 mounted on both ends of the head plate 10 are mounted, the polarities of the permanent magnets may be disposed opposite to each other and coupled to the head fixing part 20 . Opposite polarities are arranged to face each other so that the head plate 10 and the head fixing part 20 of the experimental animal are easily bound by magnetic force.

Alternatively, metal plates may be mounted on both ends of the head plate. When the metal plate is mounted on the head plate, it is made of iron, which is a metal material that can be coupled to a magnetic material, and can be manufactured in various shapes, but a circular shape is preferable.

When the experiment is finished, the head plate 10 is forcibly separated from the head plate 10 mounted on the head of the experimental animal by preventing the head plate 10 from rising by the support while the actuator 30 connecting the head fixing part 20 rises becomes

2 is a schematic diagram of an automated system for training or measuring multiple cognitive behaviors according to an embodiment of the present invention. The automation system may include a housing 100 formed of an opaque panel so as to block external light.

According to one embodiment, the housing may be made of metal or high-strength plastic, and one or more air holes may be formed at one side of the housing so that external air may flow in and internal air may be discharged to the outside.

Inside the housing, a life cage 130 in which laboratory animals live freely, a behavior experiment unit 120 connected to the life cage to perform cognitive behavioral training or experiments, and a behavior experiment unit 120 located on the front or side of the behavior experiment unit, It may include a stimulus providing unit 140 for applying the selected sensory stimulus.

It may also include a behavior detection unit 110 for applying a reward or punishment according to the cognitive behavior of the laboratory animal, and a water supply unit 180 connected to the behavior experiment unit to control water supply to the laboratory animal.

A data acquisition unit 150 that automatically measures the water intake behavior according to the multi-sensory information recognition of the laboratory animal through the signal detected by the behavior detection unit and a power supply unit that supplies electricity to circuit devices included in the housing ( 170) may be included.

In addition, the microcomputer 160 may further include training software for measuring and training the cognitive abilities of the laboratory animals, controlling operations of devices in the housing, and storing recorded data.

4 is a view showing the exterior of a housing according to an embodiment of the present invention. A water bottle 310 for supplying water is provided on one side of the outer side of the housing, and a water bottle mounting portion 320 for fixing the water bottle and a connection tube 330 for supplying water may be further included.

A touch screen 300 capable of inputting control commands, confirming test results, and monitoring the state of the test animal inside the housing in real time may be further included on one side of the outer upper surface of the housing.

The experimental animal lives freely in the life cage 130 located inside the housing, and when a movement to move to the behavior experiment unit 120 to which water is supplied is detected, the automatic shutter 40 connected to the life cage is opened. A touch sensor (not shown) is installed at the entrance of the connection unit connecting the life cage and the behavior experiment unit, and an automatic shutter is operated by the movement of the experimental animal.

When the automatic shutter 40 operates and the connection part 125 of the life cage 130 is opened, the experimental animal moves to the behavior experiment part 120 . When the entry of the experimental animal is detected in the behavior experiment unit, the actuator 30 to which the head fixing unit 20 is attached moves downward to fix the head of the experimental animal.

When training or measurement ends after a certain period of time has elapsed, the actuator 30 rises to release the restraint of the experimental animal's head. Thereafter, when detecting that the experimental animal leaves the behavior experiment unit 120 and returns to the life cage 130, the automatic shutter 40 is closed to close the connection unit 125.

6 is an example of a state in which the head fixing part 20 according to an embodiment of the present invention is fixed to the head plate 10 of the laboratory animal.

When a mouse, which is an experimental animal, enters the behavior experiment unit 130, the actuator 30 equipped with the head fixing unit 20 moves to the bottommost position. The head fixing part may include a permanent magnet or an electromagnet to be coupled with the head plate. The magnets can be placed with opposite polarities to ensure correct coupling with the headplate.

The head plate of the lab animal may be automatically bound to the head fixing part through mechanical or electromagnetic means.

When the sensor detects that the head fixing part 20 connected to the actuator 30 is coupled to the head plate 10 of the experimental animal, the actuator moves to an intermediate position and is fixed. The actuator is driven by a stepping motor and is connected to a rack and pinion gear to perform linear motion up and down.

The head-fixed experimental animal is located 1 cm in front of the behavior sensor 110 including at least one water supply port 50 and an air puff 60 . The action sensor may include at least one water supply port and an air puff for spraying air.

The water supply unit 180 may include a mini pump 182 for controlling the operation of each water supply port 50 . The mini-pump is controlled so that 4 μl of water is given as a reward when the experimental animal licks the water supply port.

7 is an example of a water compensation port 50 located in front of an experimental animal having a fixed head according to an embodiment of the present invention.

The water compensation port may be composed of at least one or more. In the case of individually controlling water supply by installing a plurality of water compensation ports, it is possible to train to distinguish various types of stimuli. The number of water compensation ports can be arranged differently depending on the training purpose.

8 is an example of a state in which a mouse being trained is monitored through an internal infrared camera 70 according to an embodiment of the present invention.

The infrared camera 70 is located at the top of one side of the behavior experiment unit 120 and can monitor the appearance of the experimental animal being trained on the external touch screen 300 . Through the infrared LED attached to one side of the infrared camera, it is possible to check the condition of the experimental animal or the progress of the experiment even in a dark environment.

9 is an example of a data server 200 that obtains a large amount of data by simultaneously connecting a plurality of automated systems for one or more multi-cognitive behavioral training or measurement according to an embodiment of the present invention.

The housing 100 of the automation system for at least one or more multi-cognitive behavior training or measurement may be stacked and fixed on the rack 220 . It can be connected to the data server 200 through wired or wireless communication through data output terminals formed on each housing. The data server may record data obtained from each system in real time. At this time, water can be supplied to the inside of each housing 100 stacked on the rack through the water tank 230 located at the upper end of the rack 220 and the water connection pipe 240.

A method for fixing the head fixing unit 20 of an automated system for training or measuring multiple cognitive behaviors according to the present invention includes the steps of moving the lab animal from the life cage 130 to the behavior experiment unit 120 by itself due to the desire to consume water, When it is detected that the experimental animal has entered the behavioral testing unit, the head fixing unit 20 descends and is automatically bound to the head plate 10 of the experimental animal, and when the training or experiment is finished according to the set training program, the head Separating the fixing part from the head plate and returning the experimental animal to the life cage after leaving the behavioral experiment part may be included.

The present invention will be described in more detail through the following examples. These examples are only for explaining the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited to these examples.

실시예Example

Multi-sensory stimulation discrimination learning of mice using the automated system for training or measuring multi-cognitive behaviors of the present invention is performed as follows.

(1) Mice between 30 and 60 days old are selected as experimental animals. Selected mice begin audiovisual stimulus discrimination training after two days of water restriction. Thereafter, water is provided only through the water compensation port 50 located in the behavior experiment unit 120 .

(2) Water reward habituation is a task for mice to learn that water comes out of the water reward port 50, and 4 μl of water is given as a reward every time the mouse licks the water reward port 1 cm away from the mouth. .

Optionally, it may be configured so that an experimenter can manually control water to come out of the water compensation port from the outside of the housing 100 .

The number of times the mouse voluntarily licks the water compensation port 50 is recorded by the 'Presentation' program built into the microcomputer 160.

(3) Stimulus conditioning is a task that allows mice to learn that they can eat water when presented with a go stimulus.

It is configured so that water comes out when the water reward port is licked within 2 seconds after the behavioral stimulus appears.

Additionally, even if the water compensation port 50 is not licked within 2 seconds, water is automatically discharged from the water compensation port, which induces the mouse to associate behavioral stimuli with water.

The number of times the mouse voluntarily licked the water reward port within 2 seconds and the number of times it did not is recorded through the behavior sensor 110, respectively, and the degree of association between the action stimulus and the water is analyzed.

External sensory stimuli are provided with visual stimulation using the monitor 142 and auditory stimulation through the speaker 141 on one side of the mouse.

(4) Stimulus discrimination training is a task in which mice learn to discriminate between go and no-go stimuli. It learns to lick the water reward port when the action stimulus comes out, and not to lick it when the flight position stimulus comes out.

To this end, water comes out only when the water compensation port 50 is licked within 2 seconds after the action stimulus appears, and if not, water does not come out. In addition, when the water compensation port 50 is licked within 2 seconds after the flight position stimulus appears, the air puff 60 applies air to the face of the mouse as punishment.

At this time, visual or auditory stimuli used as action stimuli or flight stimuli are provided randomly and given at the same frequency for 1 second.

During stimulus discrimination training, the number of times the water reward port 50 is licked for action stimuli and the number of times the water reward port is licked for flight stimuli are recorded by the computer program 'Presentation', based on which the success rate It is obtained as a behavioral index and a learning curve is drawn.

The success rate can be calculated as shown in Table 1.

	행위 자극stimulation of action	비행위 자극fly phase stimulus
핥은 경우(lick)lick	옳게 핥음(hit) = aRight lick (hit) = a	잘못 핥음(false alarm) = cfalse alarm = c
핥지 않은 경우(no lick)No lick	놓침(miss) = bmiss = b	옳게 놓침(correct reject) = dcorrect reject = d

correct rate = (a + d) / (a + b + c + d)

In the visual stimulus discrimination task, after two visual stimuli of different angles appeared on the screen for 1 second, the mouse's response to this was confirmed by whether or not the mouse licked the water reward port 50.

At this time, the stimulus that moves the 90-degree black bar to the right is used as the action stimulus, and the stimulus that moves the 180-degree black bar upward is used as the flight position stimulus.

In the auditory stimulus discrimination task, after listening to two auditory stimuli of different frequencies for 1 second, the mouse's response to this was confirmed by whether or not it licked the water reward port 50. At this time, a continuous sound of 5 kHz was used as a behavior stimulus and a continuous sound of 10 kHz was used as a flight stimulus. The decibels of all sounds were unified to 74.5 dB using a decibel meter.

The auditory stimulus may use a sine wave signal to deliver a pure tone sound. The stimulation pattern and stimulation time for visual stimulation and the frequency range and size of signals for auditory stimulation can be adjusted.

(5) a congruent audiovisual stimulus that simultaneously gives both visual and auditory stimuli corresponding to behavioral stimuli to mice trained to distinguish between visual and auditory stimuli; Mismatched audiovisual stimuli that simultaneously give an auditory stimulus and an auditory or visual stimulus corresponding to the non-congruent stimulus may be randomly given with a probability of 25% of the total. 'GO' command learning applies a 10KHz sound signal and a vertical pattern, and 'NO-GO' command learning applies a 5KHz sound signal and a horizontal pattern to learn.

In the congruent audiovisual stimulus test, which simultaneously gives visual and auditory stimuli, the learned sound stimulus and pattern pattern 'GO' command apply a 10KHz sound signal and vertical pattern pattern, and the 'NO-GO' command applies a 5KHz sound signal and a horizontal pattern.

In the mismatching audiovisual stimulus test, the 'GO' command can apply a 10KHz sound signal and a horizontal pattern, and the 'NO-GO' command can apply a 5KHz sound signal and a vertical pattern. .

10 shows results of multi-cognitive behavioral training of laboratory animals according to an embodiment of the present invention.

In a multimodal task that gives audio-visual stimulation simultaneously, it can be seen that the correct rate increases according to the learning behavior training of the normal mouse (WT) model and the autistic mouse (KO) model (FIG. 10a) . There was no difference between the behavior of the normal mouse model and the autistic mouse model in terms of the number of days it took to learn visual and auditory stimuli and the degree of discrimination ability (FIG. 10b).

In addition, in the normal mouse (WT) model and the autistic mouse (KO) model, the result of measuring the degree of visual or auditory discrimination ability that varies depending on the intensity of stimulation. It was confirmed that the autistic mouse model (open bar) had a higher ability to discriminate when the stimulus intensity was weaker than that of the normal mouse model (filled bar) (FIG. 10c).

Although described above with reference to specific embodiments of the present invention, for those skilled in the art, this is only a preferred embodiment, and the practical scope of the present invention is not limited thereto and can be modified and changed in various ways. It will be an obvious fact.

[Description of code]

housing 100 motion sensor 110

Behavior Lab 120 Life Cage 130

Stimulus supply unit 140 Data acquisition unit 150

Microcomputer 160 Power Supply 170

Water supply unit 180 Lighting unit 190

Head plate 10 Head fixing part 20

Actuator 30 Auto Shutter 40

Water Compensation Port 50 Air Puff 60

Infrared Camera 70 Data Server 200

Cable 210 Rack 220

Water Tank 230 Water Connector 240

Touchscreen 300 Water Bottle 310

Water bottle mount 320 Connecting tube 330

Claims

In an automated system for multiple cognitive behavioral training or measurement in which laboratory animals can perform behavioral training on their own,

a head plate 10 mounted on the head of an experimental animal;

a behavior experiment unit 120 in which cognitive behavioral training or experiments of laboratory animals are performed;

A head fixing part 20 located in the behavior experiment unit and coupled to the head plate 10 to fix the experimental animal; includes,

The behavior experiment unit 120 is a space separated from the space where the lab animal usually lives or a space where the lab animal can freely move and enter, and the head plate 10 is mechanically attached to the head fixing part 20. , characterized in that automatically bound through magnetic or electromagnetic means, an automated system for multiple cognitive behavioral training or measurement.
According to claim 1,

an experimental animal having the head plate 10 mounted on its head;

A housing 100 formed of an opaque panel on an entire surface to block external light;

a life cage (130) located within the housing and in which the laboratory animals freely live; and

An actuator 30 located in the behavior experiment unit 120 and connected to the head fixing unit 20 to move up and down; including,

The actuator is an automated system for training or measuring multiple cognitive behaviors, characterized in that it includes a stepping motor, a rack and pinion gear.
According to claim 2,

a stimulus providing unit 140 located on the front or side of the behavior experiment unit 120 to apply selected sensory stimuli to the experimental animal;

a behavior detection unit 110 for applying a reward or punishment according to the cognitive behavior of the laboratory animal;

A water supply unit 180 connected to the behavior experiment unit to control water supply to the experimental animals;

a data acquisition unit 150 that automatically measures a water intake behavior according to the sensory information recognition of the laboratory animal through the signal detected by the behavior detection unit;

a power supply unit 170 supplying electricity to circuit devices included in the housing;

Training software for measuring and training the cognitive abilities of the laboratory animals; and

An automated system for multi-cognitive behavioral training or measurement, further comprising; a microcomputer 160 that controls the operation of devices in the housing and stores recorded data.
According to claim 2,

The life cage 130 includes a connection part 125 connected to the behavior experiment part 120;

a touch sensor for detecting a motion of an experimental animal located in the connection unit;

an automatic shutter 40 allowing or limiting the movement of laboratory animals according to the detection of the touch sensor; and

The head plate 10 includes at least one permanent magnet or metal plate,

The head fixing part (20) further comprises at least one permanent magnet or electromagnet coupled to the head plate. An automated system for training or measuring multiple cognitive behaviors.
According to claim 2,

A water bottle 310 attached to the outside of the housing 100 and supplying water to the water supply unit;

a water bottle mounting portion 320 for fixing the water bottle to the housing; and

An automated system for multi-cognitive behavioral training or measurement, characterized in that it further comprises; a connecting tube 330 for water supply.
According to claim 3,

The action detection unit 110 includes a water compensation port 50 through which water is output for a certain period of time when the learned action is successfully recognized; and

An automated system for training or measuring multiple cognitive behaviors, characterized in that it includes an air puff (60) in which air comes out as a punishment instead of water when the learned behavior is failed to be recognized.
According to claim 2,

Characterized in that it further comprises: a touch screen 300 capable of inputting control commands to the upper surface of the housing 100, confirming experiment results, and monitoring the state of laboratory animals inside the housing in real time; An automated system for behavioral training or measurement.
According to claim 3,

The stimulus providing unit 140 includes at least one speaker 141 for applying an auditory stimulus to the lab animal; and

An automated system for multi-cognitive behavioral training or measurement, characterized in that it further comprises; at least one or more monitors 142 for applying visual stimuli.
According to claim 3,

The data acquisition unit 150 is a DAQ (Data Acquisition system) that measures the time and number of times when a behavior of the experiment animal licking the water compensation port 50 occurs in the behavior detection unit 110 and converts it into digital data. Characterized in that it further comprises, an automated system for multiple cognitive behavioral training or measurement.
According to claim 2,

Located on one side of the upper part of the behavior experiment unit 120 and further comprising an infrared camera 70 for monitoring the state of the laboratory animal even in a dark state without light inside the housing 100, multi-cognitive behavioral training or an automated system for measurement.
According to claim 2,

A lighting unit 190 mounted inside the housing 100 to provide light so that the laboratory animals can differentiate between day and night; and

A timer capable of setting the operating time of the lighting unit; characterized in that it further comprises an automated system for training or measuring multiple cognitive behaviors.
According to claim 2,

The experimental animal is an automated system for multiple cognitive behavioral training or measurement, characterized in that the experimental animal is an experimental animal with brain cognitive dysfunction or loss.
According to claim 1,

An automated system for training or measuring multiple cognitive behaviors, characterized by connecting or stacking one or more of the automated systems using a rack 220 to simultaneously train a plurality of laboratory animals or acquire cognitive ability measurement data. .
An automated method for training or measuring multiple cognitive behaviors of laboratory animals using the automated system of any one of claims 1 to 13,

(a) moving the lab animal from the life cage 130 to the behavior experiment unit 120 by itself due to the desire for water intake after water restriction;

(b) automatically binding the head fixing part 20 to the head plate 10 of the experimental animal when it is detected that the experimental animal entered the behavioral testing unit 120;

(c) separating the head fixing part 20 from the head plate 10 when training or measurement is finished according to the set training program;

(d) an automated method for multiple cognitive behavioral training or measurement, including;
According to claim 14,

The laboratory animal is characterized in that the laboratory animal model with brain cognitive dysfunction or loss, an automated method for multiple cognitive behavioral training or measurement.
According to claim 15,

The brain cognitive impairment or loss may include mild cognitive impairment, vascular cognitive impairment, cognitive impairment due to cerebral hemorrhage, dementia, brain damage, memory impairment, visual and/or auditory discrimination impairment, autism, attention deficit hyperactivity disorder , ADHD), schizophrenia or Parkinson's disease, characterized in that, an automated method for training or measuring multiple cognitive behaviors.
According to claim 15,

After step (d), further comprising a step (step e) of verifying the effect of digital therapeutics, which is a program for alleviating or treating symptoms of brain cognitive dysfunction or loss, characterized in that it further comprises multiple cognitive behavioral training or measurement. automation method for
Using the automated system of any one of claims 1 to 13, measuring changes in brain cognitive ability in laboratory animals to which a drug candidate exhibiting a preventive or therapeutic effect on brain cognitive function is administered, Brain cognition comprising the step of measuring A method for screening drug candidates that exhibit preventive or therapeutic effects on functional disorders or injuries.
According to claim 18,

The experimental animals suffered from mild cognitive impairment, vascular cognitive impairment, cognitive impairment due to cerebral hemorrhage, dementia, brain damage, memory impairment, visual and/or auditory discrimination, autism, attention deficit hyperactivity disorder (ADHD), A method for screening a drug candidate that exhibits a preventive or therapeutic effect on brain cognitive impairment or loss, characterized by having cognitive impairment or loss such as schizophrenia or Parkinson's disease.