CN109156369B

CN109156369B - Animal raising cage and method for establishing depression mouse animal model by using animal raising cage

Info

Publication number: CN109156369B
Application number: CN201811162801.2A
Authority: CN
Inventors: 田波; 张培; 祁广见
Original assignee: Huazhong University of Science and Technology
Current assignee: Huazhong University of Science and Technology
Priority date: 2018-09-30
Filing date: 2018-09-30
Publication date: 2023-07-25
Anticipated expiration: 2038-09-30
Also published as: CN109156369A

Abstract

The invention provides an animal raising cage which comprises a raising cage body (1) and a raising cage cover (2), wherein an opening (3) is formed in the upper end of the raising cage body (1), the raising cage cover (2) covers the opening (3) in the upper end of the raising cage body (1), the raising cage cover (2) is formed by transverse metal wires and longitudinal metal wires, a V-shaped structure (4) is arranged on the raising cage cover (1), and the V-shaped structure (4) is recessed towards the interior of the raising cage body (1), and the animal raising cage is characterized in that at least one raising cage partition plate (5) is arranged on the raising cage body (1), the raising cage partition plate (5) is made of transparent materials, and the raising cage partition plate (5) partitions the internal space of the raising cage body (1). The invention also provides a method for establishing a depression mouse animal model by using the rearing cage.

Description

Animal raising cage and method for establishing depression mouse animal model by using animal raising cage

Technical Field

The invention belongs to the field of basic medicine, and particularly relates to a social cage for raising animals and a method for establishing an animal disease model by using the social cage, and more particularly relates to a social cage for raising rats and mice and a method for establishing a depression mouse animal model by using the social cage.

Background

Depression is a mental disorder characterized by a major clinical depression, and so far, global patients have reached 3.4 billion, bringing great losses to individuals, families and society, but the etiology is unknown so far. Classical theory suggests that physical stress and/or emotional stress can lead to depression, but current research on pathogenesis of depression is mostly focused on molecular loop mechanisms induced by physical and emotional stress together or induced by pure physical stress, and few related researches on single pure emotional stress and depression are carried out.

At present, a mouse chronic social failure depression model is established internationally by mainly using a two-box social cage (Golden, sam A., et al, A standardized protocol for repeated social defeat stress in mice., nature protocols 6.8 (2011): 1183), a CD1 mouse with a larger size is selected to attack a C57BL/6J 6 mouse with a smaller size in a single box of the two-box social cage for 5-10 min, and then the mouse is separated and placed on the boxes at the left side and the right side, and the method combines emotional stress and physical stress in the factors inducing the depression of the mouse. In addition, a depression mouse model is built through a two-box social cage, emotion stress and somatic stress are combined in factors for inducing depression of the mouse, the method plays a vital role in the research of the related field of depression for decades, but the method cannot better simulate the inducing factors for clinical depression morbidity. Meanwhile, how to build a mouse model simulating single-pure condition induction induced depression is one of the key scientific problems to be solved urgently at present. Based on the improvement of the current widely-applied chronic social failure depression model, a novel social cage is designed to establish a depression model induced by the emotion induction, so that a research thought is provided for elucidating the pathogenesis of depression.

Disclosure of Invention

One of the purposes of the invention is to provide a novel rearing cage, which realizes the technical scheme that:

the invention provides a rearing cage which comprises a rearing cage body 1 and a rearing cage cover 2, wherein the upper end of the rearing cage body 1 is provided with an opening 3, the rearing cage cover 2 is covered on the opening 3 at the upper end of the rearing cage body 1, the rearing cage cover 2 is composed of transverse metal wires and longitudinal metal wires, the rearing cage cover 1 is provided with a V-shaped structure 4, and the V-shaped structure 4 is recessed towards the inside of the rearing cage body 1.

The lower part of the rearing cage partition board 5 can be provided with a plurality of small holes 7 which can be ventilated, so that animals on two sides of the partition board can smell odor mutually.

The rearing cage body 1 may be provided with 2 rearing cage partitions 5.

The rearing cage body 1 can be the cage body of the rearing cage of R5 type big and small mouse group, the cage cover 2 can be the cage cover of the rearing cage of R5 type big and small mouse group, the rearing cage body 1 be provided with 2 rearing cage baffle plates 5, rearing cage baffle plate lower part open and have a plurality of apertures 7.

Preferably, the wide side of the cage body is equally divided into three parts by 2 cage partitions 5, the cage partitions 5 are made of transparent resin materials, the cage partitions are clamped by fixing plates 8, the fixing plates 8 are fixedly connected to the side walls of the cage, and the fixing plates 8 are provided with a strip-shaped groove 9, so that two ends of the cage partitions 5 can slide up and down along the strip-shaped groove 9, and can be taken out or put in.

It is another object of the present invention to provide a method for establishing a mouse animal model of depression, comprising the steps of:

(1) Placing a C57BL/6J witness mouse, a CD1 (ICR) challenged mouse and a C57BL/6J challenged mouse in sequence in three spaces of the rearing cage which are separated by the rearing dragon baffle plate;

(2) Challenged mice were placed daily in the grid where CD1 (ICR) challenged mice were placed, and challenged for 3-15 minutes before being returned to the original grid for a total of 5-15 days.

Preferably, the challenge in step (2) is 5 minutes for a duration of 10 days;

preferably, step (2) is followed by step (3) and step (4) of checking the modeling effect:

(3) Detecting open field, overhead plus maze, tail suspension and forced swimming of the mice on the next day after the lasting days of the step (2);

(4) The sugar water preference experiment is detected the next day after the end of step (3).

The rearing cage provided by the invention has obvious effects in building a single pure condition induction induced depression mouse model by combining with the modeling method provided by the invention, and has multiple advantages compared with the existing modeling method: the rearing cage provided by the invention can be used for simultaneously establishing the emotion stress and somatic stress group model and the simple emotion stress group model. In the traditional method, the two models are required to be respectively established, the steps are complicated to be respectively established, and systematic errors are easy to generate when the models of the emotion induction stress group, the body stress group and the simple emotion stress group are respectively established. The invention can simultaneously establish the two models, can eliminate systematic errors while reducing modeling operation flow, can better simulate the inducing factors of clinical depression onset, is a powerful tool for researching single-pure condition induction induced depression molecules and nerve loop mechanisms, and can provide a new animal model for depression drug treatment and evaluation.

Drawings

FIG. 1 is a view showing the whole of the cage of example 1 at different angles: including top view (fig. 1A), diagonal top view (fig. 1B), left wide side top view (fig. 1C), and left wide side top view (fig. 1D);

FIG. 2 is a view of the feeder cage of example 1 in a non-covered physical form at different angles: including top view (fig. 2A), long side top view (fig. 2B), and left wide side top view (fig. 2C);

FIG. 3 is a cage divider plate of example 1: a physical diagram (figure 3A) and a structural schematic diagram (figure 3B) of the transparent resin partition board with the holes;

FIG. 4 is a physical view (FIG. 4A) and a schematic view (FIG. 4B) of a fixing plate of a cage partition;

fig. 5 is a general flow chart for establishing a single-pureness induction-induced depression mouse model: overall flow chart (fig. 5A), three groups of mice treatment details (fig. 5B);

FIG. 6 is a graph showing the results of behavioral tests of a control group, a simple stress group, an affective stress group and a somatic stress group after establishing a mouse depression animal model according to the method provided in example 2;

FIG. 7 is a schematic view of the structure of the feeder cage provided by the invention;

FIG. 8 is a schematic view of the structure of the cage partition provided by the invention;

fig. 9 is a schematic view of a structure of a fixing plate of a partition plate of a rearing cage provided by the invention.

Detailed Description

The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention. In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.

Example 1:

the embodiment provides a raising cage based on improvement of R5 type big and small mouse group raising cage, namely a novel three-box social cage of a novel three-box social cage, which consists of an R5 type big and small mouse group raising cage, a transparent resin partition plate with holes and a transparent resin partition plate fixing plate with holes. Fig. 1 is a novel three-box social cage overall physical diagram at different angles, and mainly comprises overlooking (fig. 1A), diagonal overlooking (fig. 1B), left wide overlooking (fig. 1C) and left wide overlooking (fig. 1D). Fig. 2 is a novel three-box social cage uncovered physical diagram at different angles, including top view (fig. 2A), long side top view (fig. 2B) and left wide side top view (fig. 2C). Fig. 3 is a physical diagram (fig. 3A) and a structural schematic diagram (fig. 3B) of a perforated transparent resin partition plate, fig. 4 is a physical diagram (fig. 4A) and a structural schematic diagram (fig. 4B) of a perforated transparent resin partition plate fixing plate, fig. 3 and fig. 4 are main design parts of the raising cage according to the invention, two perforated transparent resin partition plates equally divide the wide side of the raising cage for the R5 type mice into three parts, the partition plates are made of transparent resin materials, specific structures and parameters are shown as fig. 3B, the partition plates are clamped by fixing plates and can be put in or taken out from top to bottom, and the fixing plates are also made of transparent resin materials, and specific structures and parameters are shown as fig. 4B. Electric drills are adopted for punching the partition plates and the fixing plates, each partition plate is punched 5 multiplied by 24, the diameter is 4-6mm, each fixing plate is composed of two single plates, each single plate is punched 2, the diameter is 4-6mm, and the corresponding screws are selected for fixing.

Example 2:

this example is a single pureness induction induced depression mouse model established in accordance with the method of establishing a depression mouse animal model provided by the present invention. The general flow chart is shown in fig. 5A. Firstly, placing C57BL/6J witness mice (simple emotion stress group), CD1 challenged mice and C57BL/6J challenged mice (emotion stress and somatic stress group) in a novel three-box social cage from left to right in sequence, putting challenged mice into a grid where the CD1 mice are located for 5min and then placing the challenged mice back into the original grid for 10 days; and after the modeling is finished, detecting the open field, the overhead plus maze, the tail suspension and the forced swimming of the mice on the 11 th day, and starting the syrup preference experiment on the 12 th day. The control groups were C57BL/6J mice, empty, C57BL/6J mice in this order, and the three groups of mice were treated in a specific manner as shown in FIG. 5B.

Statistical analysis is carried out by adopting unpaired t test in all behavioral experimental results, and ns is totally called no signalicant, which indicates that p is more than or equal to 0.05; * Represents p less than 0.05; * I.e. p is less than 0.01; * P is shown to be less than 0.001. In addition, the number of mice in the control group, the simple emotion stress group and the three groups of emotion stress and somatic stress group is 21, 31 and 31 respectively. The social experiment result shows that compared with the control group, the social ratio difference of the simple emotion stress group has no statistical significance, the difference of the emotion stress group and the somatic stress group has statistical significance, and meanwhile, the difference of the emotion stress group and the somatic stress group and the single pure emotion stress group has significant statistical significance. In the open field experiment, the results of the retention time of the central area (fig. 6B) and the movement distance of the central area (fig. 6C) show that the difference between the single pure emotion induction group and the control group has a statistical significance, and the emotion induction group and the somatic stress group have a significant statistical significance compared with the control group, but have no difference from the single emotion induction group. In the overhead plus maze experiment, the results of the open arm retention time (fig. 6D), the open arm movement distance (fig. 6E) and the open arm entry times (fig. 6F) all show that compared with the control group, the simple emotion stress group and the emotion stress and somatic stress group are obviously reduced, and meanwhile, no obvious difference exists between the single pure emotion stress group and the emotion stress and somatic stress group. The immobility time in the tail suspension experiment (fig. 6G) and the immobility time in the forced swimming experiment (fig. 6H) are significantly higher than those in the control group and the simple emotion stress group, and meanwhile, there is no difference between the control group and the simple emotion stress group. In the sugar water preference experiment, the preference percentage (fig. 6I) was significantly lower in the affective and somatic stress groups than in the control group and the simple affective group.

It will be readily appreciated by those skilled in the art that the foregoing description is merely a preferred embodiment of the invention and is not intended to limit the invention, but any modifications, equivalents, improvements or alternatives falling within the spirit and principles of the invention are intended to be included within the scope of the invention.

Claims

1. A method for establishing a depressive mouse animal model, which is characterized in that a cage is used for establishing the depressive mouse animal model, the cage comprises a cage body (1) and a cage cover (2), the upper end of the cage body (1) is provided with an opening (3), the cage cover (2) covers the opening (3) at the upper end of the cage body (1), the cage cover (2) is formed by transverse wires and longitudinal wires, the cage cover (2) is provided with a V-shaped structure (4), the V-shaped structure (4) is recessed towards the interior of the cage body (1), the cage body (1) is provided with 2 cage partition boards (5), the cage partition boards (5) are made of transparent materials, the inner space of the cage body (1) is separated by the cage partition boards (5), one side of the upper edge of the cage partition boards (5) is provided with a V-shaped structure (4) corresponding to the cage cover (2), the recessed part (6) is arranged on the corresponding to the V-shaped structure (4), and the recessed part (6) can not be matched with the cage cover (2) when the V-shaped structure (4) is covered by the cage cover (2), so that the animal structures (2) can be covered by the cage cover (2) or can be covered by the cage cover;

the method comprises the following steps:

(1) In the three spaces separated by the rearing cage partition plate, a C57BL/6J witness mouse, a CD1 (ICR) challenged mouse and a C57BL/6J challenged mouse are placed in sequence;

(2) Challenged mice were placed daily in the grid where CD1 (ICR) challenged mice were placed, and after challenged, returned to the original grid.

2. The method for constructing a model of a depressive mouse according to claim 1, wherein the lower part of the cage partition (5) is provided with a plurality of small holes (7).

3. The method for establishing a depressive mouse animal model according to claim 1, wherein the rearing cage body (1) is a cage body of an R5 type rat group rearing cage, the rearing cage cover (2) is a cage cover of an R5 type rat group rearing cage, the rearing cage body (1) is provided with 2 rearing cage partition plates (5), and a plurality of small holes (7) are formed in the lower portion of the rearing cage partition plates.

4. A method for building an animal model of depressed mice according to claim 3, wherein the wide side of said R5-type mouse group cage body is divided equally into three parts by 2 said cage partitions (5), said cage partitions (5) are made of transparent resin material, said cage partitions are held by fixing plates (8), said fixing plates (8) are fixedly attached to the side walls of the cage, said fixing plates (8) are provided with a bar-shaped groove (9) so that both ends of said cage partitions (5) can slide up and down along said bar-shaped groove (9) to be taken out or put in.

5. The method for constructing an animal model of depression mice according to claim 1, wherein in the step (2), the challenged mice are placed in the cell where the CD1 (ICR) challenged mice are located every day, and are returned to the original cell after being challenged for 3 to 15 minutes for 5 to 15 days.

6. The method for constructing a mouse animal model of depression according to claim 1, wherein the challenge time in step (2) is 5 minutes and the duration is 10 days.

7. The method for constructing a mouse animal model for depression according to claim 1 or 6, wherein step (2) is followed by step (3) and step (4) of examining the modeling effect: