CN106982740B - Experimental device for establish anxiety animal model - Google Patents

Experimental device for establish anxiety animal model Download PDF

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Publication number
CN106982740B
CN106982740B CN201611265152.XA CN201611265152A CN106982740B CN 106982740 B CN106982740 B CN 106982740B CN 201611265152 A CN201611265152 A CN 201611265152A CN 106982740 B CN106982740 B CN 106982740B
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box body
animal model
stimulation
anxiety
establishing
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CN106982740A (en
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邵杰
杨帆
屠洁
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Shenzhen Biological Manufacturing Industry Innovation Center Co.,Ltd.
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Shenzhen Institute of Advanced Technology of CAS
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K1/00Housing animals; Equipment therefor
    • A01K1/02Pigsties; Dog-kennels; Rabbit-hutches or the like
    • A01K1/03Housing for domestic or laboratory animals
    • A01K1/031Cages for laboratory animals; Cages for measuring metabolism of animals
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K29/00Other apparatus for animal husbandry
    • GPHYSICS
    • G05CONTROLLING; REGULATING
    • G05BCONTROL OR REGULATING SYSTEMS IN GENERAL; FUNCTIONAL ELEMENTS OF SUCH SYSTEMS; MONITORING OR TESTING ARRANGEMENTS FOR SUCH SYSTEMS OR ELEMENTS
    • G05B19/00Programme-control systems
    • G05B19/02Programme-control systems electric
    • G05B19/04Programme control other than numerical control, i.e. in sequence controllers or logic controllers
    • G05B19/042Programme control other than numerical control, i.e. in sequence controllers or logic controllers using digital processors

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Zoology (AREA)
  • Clinical Laboratory Science (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Engineering & Computer Science (AREA)
  • Automation & Control Theory (AREA)
  • Housing For Livestock And Birds (AREA)

Abstract

The invention relates to the technical field of experimental instruments, in particular to an experimental device for establishing an anxiety animal model. The experimental device for establishing the anxiety animal model comprises a box body, wherein a feeding device, a control system and a stimulus source are arranged in the box body, and the stimulus source is electrically connected with the control system and works under the control of the control system. According to the invention, the feeding device and the stimulation source are arranged in the box body, so that the feeding and stimulation experiments of animals can be realized at the same position, and the purpose of long-term experiment can be further realized; and the stimulation source is controlled by the control system, so that the stimulation automatic driving to a certain degree is realized.

Description

Experimental device for establish anxiety animal model
Technical Field
The invention relates to the technical field of experimental instruments, in particular to an experimental device for establishing an anxiety animal model.
Background
During the long-term evolution, many animals develop an emotional response of "anxiety" to respond to external stressors, which is compatible with the fact that external stimuli are realistic, and which is beneficial for the organism to mobilize its own potential and resources to respond to realistic stressors. For example: when the animal perceives the presence of predators, the hormone levels in the body rise, the muscles contract, and ready to escape. Therefore, appropriate levels of anxiety are beneficial to the individual, the emotional response is relieved as the environmental stress is removed, and the individual's adaptability to the particular stress environment is improved, but long-term anxiety can cause adverse effects, emotional and physiological responses can exist for a long time, and mental diseases can be induced. With the pace of modern human life increasing, there are various stimuli in daily life including work and family stress and even some negative news messages. Under these excitatory "bombings", modern populations with high living pressures become more anxious, and long term past anxiety affects brain outcome and function, thereby manifesting as mental disorder and inducing or exacerbating a range of other mental disorders including depression, schizophrenia and post-traumatic stress disorder, among others. Therefore, establishing an anxiety animal model to explore the influence of the anxiety reaction on the brain structure and the brain function becomes a hot point of research of people.
Factors that induce anxiety in animals include environmental factors and individual factors. The environmental factors are mainly external pressure stimulation, which can be very intense short-time stimulation or long-time chronic mild stimulation. The greater the likelihood that an individual will develop anxiety when an external stress stimulus becomes unpredictable. When the intensity of the stress stimulus is further increased to become non-defeatable, it is possible to induce a emotional response of "depression". Individual factors mainly refer to the sensitivity of an individual to external stress, and are mainly influenced by the experience of inheritance and key stages of development. In general, biological individuals sensitive to external stress are very susceptible to pathological anxiety emotional reactions upon prolonged unpredictable stress stimuli.
The existing experimental scheme for establishing the animal anxiety model mainly comprises two types: one is to alter brain structure by pharmacological means or physical injury, thereby impairing the brain's regulatory function in anxiety, but the subsequent effects of such methods are complex; another is to expose the animal to chronic stress stimuli or brief intense stimuli, the nature of which can be mixed singly or in combination, ultimately producing an anxiety response based on self-experience acquired habits. The more mature experimental paradigm has binding, crowding, forced swimming, predator threats, social pressure, and the like. The implementation of these single paradigms requires corresponding laboratory equipment, resulting in waste of laboratory equipment and space resources. And anxiety modeling can be performed only on a single animal at a time, and a great amount of precious time is consumed by experimental workers.
There are attempts to induce anxiety responses using relatively simple stimuli, such as an environmental stimulus control circuit optimally designed in the 200820154038.4 patent application, which provides a current stimulus of 0-2.5mA, a sound stimulus of 0-130dB, and a light stimulus of 0-45W. Patent application No. 201520126445.4 may mix current stimuli, light stimuli of different intensities generated by LED lights, and noise stimuli generated by speakers to induce the generation of an anxiety response. And the scheme uses a programmable logic device to generate random data to control the driving of the three stimuli, thereby realizing the unpredictability of the stress.
The existing experimental device for inducing the animal to generate the anxiety response by the stimulation of the environmental pressure has certain defects. For a single experimental paradigm model building, such as crowding, predator threats and the like, experimental personnel are required to erect corresponding behavioural equipment for each implementation; when a plurality of paradigms are required to be mixed for use, more equipment is required. Nowadays, behavioral experiments are mostly carried out in a barrier environment, space resources are precious, and the scheme wastes manpower and space resources greatly in a laboratory.
The main drawback of this technique for inducing an anxiety response using a single stimulus is that the single electrical, acoustic, and optical stimuli are artificially generated and do not truly mimic the environmental stress stimuli in nature. Factors inducing the animals to generate anxiety in the natural environment are mostly living environments with severe conditions, such as dampness and crowding; or the presence of a natural enemy or an intruder who is not warriorable, etc. Therefore, the animal model of anxiety built by the scheme has difficulty in convincing the similarity with animals with anxiety in natural environment. In addition, these devices can only be used for a short time, and after a single experiment is finished, the animals still need to be transferred to a breeding room, and long-term experiments cannot be carried out.
Disclosure of Invention
The invention aims to provide an experimental device for establishing an anxiety animal model, which can solve the problem that the existing experimental device cannot carry out long-term experiments.
In order to achieve the purpose, the invention adopts the following technical scheme:
an experimental device for establishing an anxiety animal model comprises a box body, wherein a feeding device, a control system and a stimulus source are arranged in the box body, and the stimulus source is electrically connected with the control system and works under the control of the control system.
As a preferable embodiment of the experimental device for establishing the animal model of anxiety, the stimulus source includes an electrical stimulation device, a sound stimulation device and a water spraying device.
As a preferable mode of the experimental device for establishing the animal model of anxiety, the electrical stimulation device includes a coil disposed at the bottom of the case, and the coil and the bottom plane of the case have a certain gap.
As a preferable scheme of the experimental device for establishing the anxiety animal model, padding is arranged at the bottom of the box body, and the thickness of the padding is adjustable.
As a preferable embodiment of the experimental device for establishing the animal model of anxiety, the coil is wound to form a plurality of rectangular frames.
As a preferable scheme of the experimental device for establishing the animal model of anxiety, the sound stimulation device comprises a voice player arranged in the box body.
As a preferable scheme of the experimental device for establishing the animal model of anxiety, the water spraying device comprises a spraying head arranged in the box body, and the spraying head is connected with the water supply device through a pipeline;
the spraying device also comprises a control valve used for controlling the water supply device to supply water for the spraying head.
As a preferable scheme of the experimental device for establishing the anxiety animal model, the feeding device comprises a feed cage and a drinking bottle which are arranged in a box body;
the drinking bottle comprises a drinking nozzle, and a stainless steel ball is arranged at the water outlet end of the drinking nozzle.
As a preferable scheme of the experimental device for establishing the anxiety animal model, the feeding device further comprises a ventilation device, the ventilation device comprises an air inlet pipeline communicated with the box body, and a filter is arranged in the air inlet pipeline.
As a preferable scheme of the experimental device for establishing the anxiety animal model, the control system comprises a control system composed of a single chip microcomputer, a control circuit and an operation panel, wherein the operation panel comprises a timing switch 302, a stimulation mode setter, a circulation setter and a power switch.
The invention has the beneficial effects that: according to the invention, the feeding device and the stimulation source are arranged in the box body, so that the feeding and stimulation experiments of animals can be realized at the same position, and the purpose of long-term experiment can be further realized; and the stimulation source is controlled by the control system, so that the stimulation automatic driving to a certain degree is realized.
Drawings
FIG. 1 is a schematic diagram of an experimental apparatus for establishing an animal model of anxiety according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of an experimental apparatus for establishing an animal model of anxiety according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a control panel according to an embodiment of the present invention;
FIG. 4 is a side view of an experimental setup for establishing an animal model of anxiety according to an embodiment of the present invention;
fig. 5 is a top view of an experimental apparatus for establishing an anxiety animal model according to an embodiment of the present invention.
101: a control system; 102: an electrical stimulation device; 103: a sound stimulation device; 104: a water spraying device; 105: a feeding device; 106: a ventilation device;
301: an operation panel; 302: a timing switch; 303: a stimulation mode setter; 304: a cycle setter; 305: a power switch; 306: a coil; 307: a voice player; 308: a shower head; 309: an electric ball valve; 310: a feed cage; 311: a drinking bottle; 312: a filter; 313: an SD card; 314: a front bulkhead; 315: a rear bulkhead; 316: a water pipe; 317: a drinking nozzle.
Detailed Description
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
As shown in fig. 1 to 5, in the present embodiment, an experimental apparatus for establishing an anxiety animal model is provided, which includes a box body, a feeding device, a control system 101 and a stimulation source are disposed in the box body, wherein the stimulation source is electrically connected to the control system 101 and operates under the control of the control system 101.
In the embodiment, the feeding device and the stimulation source are arranged in the box body, so that the feeding and stimulation experiments of animals can be realized at the same position, and the purpose of long-term experiments can be further realized; and the stimulation source is controlled by the control system 101 to realize the automatic driving of the stimulation to a certain degree.
Specifically referring to fig. 1, the stimulus sources include an electrical stimulation device 102, a sound stimulation device 103 and a water spraying device 104, and through the arrangement of a plurality of stimulus sources, the experimental device integrates a plurality of environmental stress stimulation conditions such as crowding, dampness and predator threats, and establishes a more reliable animal model of anxiety. The feeding means comprise feeding means 105 and ventilation means 106. The system can be connected with 220V commercial power for power supply, the feeding device 105 only needs to place drinking water and grains at proper positions, and animals can obtain the water automatically without power supply and control.
Referring to fig. 2, 4 and 5, the present embodiment further provides a specific structure of the experimental apparatus, and the whole experimental apparatus is designed based on the box body of IVC rat cage with the common size (330mm × 205mm × 180mm) on the market, and the size of the box body is 330mm × 205mm × 130 mm. The box body is internally provided with a front clapboard 314 and a rear clapboard 315, the space in the box body is divided into a front area, a middle area and a rear area by the front clapboard 314 and the rear clapboard 315, wherein the control system 101 is arranged in the front area, the feeding device is mainly arranged in the rear area, and the middle area is a big mouse moving area. Referring to fig. 3, the control system 101 is composed of a single chip microcomputer, a control circuit and an operation panel 301, and the operation panel 301 includes a timing switch 302, a stimulation mode setter 303, a cycle setter 304 and a power switch 305. Through the timing switch 302, the stimulation mode setter 303 and the cycle setter 304 can set the stimulation mode, time, sequence, etc. to control the corresponding circuits to generate stimulation.
The electrostimulation device 102 comprises a coil 306 arranged at the bottom of the box body, a certain gap is formed between the coil 306 and the bottom plane of the box body, meanwhile, padding is arranged at the bottom of the box body, the thickness of the padding is adjustable, and the space at the lower part of the coil can be adjusted through adjusting the thickness of the padding. The sound stimulation device 103 comprises a speech player 307 arranged within the housing. The water spraying device 104 is mainly composed of a shower head 308 and an electric ball valve 309. The feeding device comprises a feeding device 105 and a ventilation device 106, wherein the feeding device 105 consists of a feed cage 310 and drinking bottles 311; the ventilation unit 106 includes an air inlet duct communicating with the housing, a filter 312 is provided in the air inlet duct, an outlet end of the air inlet duct is positioned in a middle active area of the housing and supplies fresh air to rats and mice, and the inlet end is connected to ventilation equipment of the animal room.
During the whole molding process, animals are placed at the bottom of the cage paved with the padding. For the control system 101, the whole system is controlled by the single chip microcomputer, and the purpose of controlling the time period of stimulation given by the stimulation device every day can be achieved by changing the position of the shifting sheet of the timing switch 302 on the dial plate; the stimulation mode setting device 303 can set the type and sequence of each stimulation, the numbers on the interface correspond to the sequence of the stimulation, and the '+' or '-' key is used to select from four types of electrical stimulation, sound stimulation, moist stimulation and no stimulation, and the default time of each stimulation is 10 min; the cycle setter 304 can set the number of times of the stimulation mode cycle operation, the set number of times cannot exceed the number of times allowed by the time of the opening of the timing switch, and the exceeding part is ignored; the power switch 305 controls the power supply of the entire apparatus. When the stimulation control circuit runs, the single chip microcomputer receives information such as stimulation modes, time and sequences set by the operation panel 303 and controls corresponding circuits to generate stimulation.
When the single chip microcomputer activates the electric stimulation device 102 through the control circuit, the coil 306 is electrified with a constant current of 1mA, animals can limit the action range below the coil due to electric shock stimulation of the coil, the crowding effect is achieved, and the space below the coil can be reduced in a padding adding mode. Without electrical stimulation, the animal may move past the coil 306. The coil 306 is made of stainless steel, the overall size is 20cm × 20cm, 16 small boxes of 5cm × 5cm are arranged inside the coil, and the height of the padding is not higher than that of the coil 306.
The sound stimulation device 103 is mainly constituted by a voice player 307. The device can play the voice file in the memory card after being activated, the memory card is a pluggable SD card 313, and the device can automatically download and store the sound of the natural enemy of the modeling animal and does not change the sound regularly, so that the animal is prevented from adapting to a certain sound. The speaker is positioned 10cm above the bottom of the cage and behind the front baffle 314. The sound stimulation time is 10min each time, and the voice player can play the audio files in the memory card in a circulating mode all the time. The whole box body is made of Polysulfone (PSU) plastic and can be disinfected by acid-base detergent. The joint of the plastic cover and the box body is sealed by a rubber ring, so that the influence of internal sound on surrounding animals is avoided.
In the case of the water sprinkler 104, the water sprinkler 104 is directly connected to the drinking bottle 311, and water is supplied from a water supply pipe. Specifically, the spray header 308 is connected with a water supply device through a pipeline, the water supply device is a drinking bottle 311, the drinking bottle 311 is the same as the drinking bottle 311 in the feeding device 105 described below, the drinking bottle 311 comprises a drinking nozzle, a stainless steel ball is arranged at the water outlet end of the drinking nozzle, the drinking bottle 311 is a sealed container, the water inlet end of the drinking bottle is communicated with a water supply pipeline, and an electric ball valve 309 is arranged on the water supply pipeline. When the water spraying device 104 is activated, the control power supply of the electric ball valve 309 can be switched on, water is introduced into the drinking bottle 311 through the water pipe 316 interface, and after the water level rises to the height of the spray header 308, water can be sprayed out from the spray header 308 by the water pressure, so that the hair of animals can be wetted, and a humid environment can be simulated. The spray header 308 is 10cm higher than the bottom of the cage. The stainless steel ball is arranged in the top of the drinking nozzle of the drinking bottle 311, and the stainless steel ball blocks the passage of the drinking nozzle due to water pressure, so that water can only be sprayed out from the spray header 308. In 10min of activation of the device, the electric ball valve 309 fills the drinking bottle only in the first 1min, then the water is sprayed out from the spray head 308, the water level is reduced below the spray head 308 in the remaining time, the water spraying amount is controlled within 100ml each time, and excessive water in the tank is avoided. In addition, every 2-3 days the water saturated bedding should be changed to fresh, dry bedding.
The animals may receive feed and drinking water via the feeding device 105. Solid feed can be placed in the feed cage 310, and the height of the bottom of the feed cage 310 is 5cm, so that animals can easily obtain the feed only by standing vertically. The top of the drinking nozzle 317 is provided with a stainless steel ball, animals can obtain drinking water by licking the stainless steel ball, and the drinking bottle 311 is as high as the feed cage 310. In the whole molding process, the filter 312 filters and exhausts gas in the device and introduces fresh air, and the filter element is composed of silica gel and activated carbon and mainly adsorbs water and ammonia in the air.
In order to further explain the experimental apparatus, the present embodiment further provides a specific experimental flow of the experimental apparatus:
1) replacing fresh bedding, feed and drinking water, placing animals into the device, sealing and connecting into a ventilation system;
2) turning on a power switch 305 of the device, adjusting a timing switch 302 to set the time of stimulus administration per day, and setting a stimulus mode by a stimulus mode setter 303 and a cycle setter 304;
3) changing the padding every 2-3 days, changing a timing switch 302, a stimulation mode setter 303 and the like, and changing the time, the type and the sequence of stimulation;
4) animals were removed 3-4 weeks later and the animal was monitored for anxiety using a behavioral method.
It should be noted that: the specific structural form of the stimulus sources provided in this embodiment is only for explaining the working process of the experimental device, and corresponding stimulus sources can be selectively added or replaced according to actual needs, for example, the invention can be used for experimental research such as fear, depression and the like in addition to producing long-term chronic stimulus-induced anxiety response. If the electric shock coil is replaced by a more compact fence structure so that the animal is always positioned above the coil, and the intensity and time of various types of stimulation are increased, the scheme is easier to induce a depression model.
If the camera is added, the difference of behaviors of the animal individuals in response to external pressure stimulation and the change of the behaviors of the animal individuals in the whole modeling process can be recorded in real time, the influence of the stimulation on the behaviors of the animals can be known in more detail, and more powerful evidence is provided for the reliability of the anxiety modeling paradigms.
The technical principle of the present invention is described above in connection with specific embodiments. The description is made for the purpose of illustrating the principles of the invention and should not be construed in any way as limiting the scope of the invention. Based on the explanations herein, those skilled in the art will be able to conceive of other embodiments of the present invention without inventive effort, which would fall within the scope of the present invention.

Claims (7)

1. The experimental device for establishing the anxiety animal model is characterized by comprising a box body, wherein a feeding device, a control system (101) and a stimulus source are arranged in the box body, wherein the stimulus source is electrically connected with the control system (101) and works under the control of the control system (101); the stimulation source comprises an electrical stimulation device (102), a sound stimulation device (103) and a water spraying device (104), the electrical stimulation device (102) comprises a coil (306) arranged at the bottom of a box body, a certain gap is formed between the coil (306) and the bottom plane of the box body, padding is arranged at the bottom of the box body, the thickness of the padding is adjustable, and when the coil is electrified, the behavior range of an animal to be tested is located below the coil.
2. The experimental setup for establishing an animal model of anxiety according to claim 1, characterized in that the coil (306) is coiled to form a plurality of rectangular boxes.
3. The experimental setup for establishing an animal model of anxiety according to claim 1, characterized in that the sound stimulation device (103) comprises a voice player disposed in a housing.
4. The experimental device for establishing the animal model of anxiety according to claim 1, wherein the water spraying device (104) comprises a spray header (308) arranged in a box body, and the spray header (308) is connected with a water supply device through a pipeline;
the device also comprises a control valve used for controlling the water supply device to supply water for the spray header (308).
5. The experimental setup for establishing an animal model of anxiety according to claim 1, characterized in that the feeding device comprises a feed cage (310) and a drinking bottle (311) arranged in a box body;
the drinking bottle (311) comprises a drinking nozzle (317), and a stainless steel ball is arranged at the water outlet end of the drinking nozzle (317).
6. The experimental apparatus for establishing the animal model of anxiety according to claim 1 or 5, wherein the feeding device further comprises a ventilation device (106), the ventilation device (106) comprises an air inlet duct communicated with the box body, and a filter (312) is arranged in the air inlet duct.
7. The experimental device for establishing the animal model of anxiety according to claim 1, wherein the control system comprises a control system (101) composed of a single chip microcomputer, a control circuit and an operation panel (301), and the operation panel (301) comprises a timing switch (302), a stimulation mode setter (303), a cycle setter (304) and a power switch (305).
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