Device and method for in vivo comparison of changes in neuronal activity in specific brain regions during odor valence changes in mice

Abstract

The present invention provides an apparatus and method for comparing changes in neuronal activity in specific brain regions of mice during the process of odor valence change in vivo, and belongs to the field of animal experiment technology. The apparatus of the present invention pre-embeds a calcium signal acquisition device in a specific brain region of mice and pre-injects a calcium ion indicator, sets a first region, a transition region, and a second region in a multi-channel odor condition preference device with controllable delivery, and during the experiment, different odors are introduced into the first region and the second region, and the movement trajectory and residence time of the mice in the first region and the second region are recorded by a movement trajectory recording system. The neuronal calcium signal acquisition system acquires and analyzes calcium signals in specific brain regions of mice, judges the valence and valence change of the mice to the odor, and uses the attraction, avoidance, or residence behavior of the mice as a behavioral representation of the odor valence, and associates it with the neuronal calcium signals in specific brain regions, thereby improving the accuracy of the association without affecting the free movement of the mice in a limited activity area.

Description

Technical Field

[0001] The present invention relates to the field of animal experiment technology, and in particular to a device and method for comparing changes in neuronal activity in specific brain regions during odor valence transformation in mice in vivo. Background Art

[0002] As a carrier of information, the information it carries can be innate, acquired, or produced with the help of chemical substances. Odors of different characteristics carry the same or different information. The information carried by odor molecules can be collectively referred to as the valence of the odor. The olfactory bulb is the relay station for odor information processing. After the action potential signal mediated by the odor molecules is transmitted to it, it continues to be transmitted deep into the brain through a large number of axonal and dendritic branches. After being encoded and processed by the primary olfactory cortex, the odor information is transmitted to the midbrain limbic system composed of brain regions such as the hippocampus, entorhinal area, dentate gyrus, and amygdala to form different odor valences. The connection between odor and odor valence is close and complex.

[0003] Therefore, when studying the relationship between odor and odor valence and the neural mechanisms of multiple brain regions, mice need to be exposed to odors with different characteristics in one experiment. However, mice cannot communicate with humans at the language level, so they can only characterize different valences through specific behaviors when exposed to odors: for example, when mice smell a favorite odor, they show attraction behavior, and when they smell a disgusting or fearful odor, they show avoidance behavior. These attraction, avoidance or staying behaviors are collectively referred to as the mouse's representation of different odor valences. Therefore, the collection and comparative analysis of neuronal activity signals in different brain regions is the key to analyzing the relationship between odor and odor valence.

[0004] The transmission of nerve signals depends on the flow of calcium ions in neuronal cells. Intracellular calcium ions are important signal molecules, and their effects are temporal and spatial. When the first cell is excited, an electrical impulse is generated. At this time, calcium ions outside the cell flow into the cell, prompting the cell to secrete neurotransmitters. Neurotransmitters bind to protein molecules on the membrane of the next-level nerve cells, prompting the nerve cells at this level to generate new electrical impulses. By analogy, nerve signals are transmitted level by level, thus forming a complex signal system and ultimately forming the advanced functions of the brain.

[0005] With the development of the times and the rapid advancement of neurobiological technology, the technology of real-time in vivo acquisition of changes in neuronal calcium activity in specific brain regions has emerged. This technology uses special fluorescent dyes or calcium ion indicators to express changes in calcium ion concentration in neurons through fluorescence intensity to reflect neuronal activity, thereby enabling the use of specific fiber optic patch cords to acquire neuronal calcium signals in specific brain regions in real time in vivo while experimental animals are awake and freely moving.

[0006] However, current fiber optic signal acquisition systems focus more on in vivo acquisition and reduction of signal-to-noise ratio, and are unable to address the need to simultaneously acquire signals of changes in neuronal calcium activity in specific brain regions with high temporal precision during odor valence representation and valence conversion. Summary of the invention

[0007] In order to solve the technical problems existing in the prior art, the present invention provides a device and method for comparing the changes in neuronal activity in specific brain regions of mice during the transition of odor valence in vivo. The device pre-embeds a calcium signal acquisition device in a specific brain region of mice and pre-injects a calcium ion indicator, and sets a first area, a transition area, and a second area in a multi-channel odor conditional preference device with controllable delivery. During the experiment, different odors are introduced into the first area and the second area, and the movement trajectory and residence time of the mice in the first area and the second area are recorded by the movement trajectory recording system. The neuronal calcium signal acquisition system acquires and analyzes the calcium signals of the specific brain regions of the mice, determines the valence and valence transition of the mice to the odor, and uses the attraction, avoidance or residence behavior of the mice as the behavioral representation of the odor valence, which is associated with the neuronal calcium signals in the specific brain regions, thereby improving the accuracy of the association without affecting the free movement of the mice in the limited activity area.

[0008] The present invention provides a device for comparing changes in neuronal activity in specific brain regions during the process of odor potency transformation in mice in vivo, comprising a controllable delivery multi-channel odor conditional preference device, a motion trajectory recording system, and a neuronal calcium signal acquisition system; during acquisition, the mouse is placed in the controllable delivery multi-channel odor conditional preference device;

[0009] The controllable delivery multi-channel odor condition preference device comprises at least three areas, a first area, a second area and a transition area; different odors can be introduced into the first area and the second area, and the first area, the second area and the transition area can be connected through control;

[0010] The motion trajectory recording system records the motion trajectory and residence time of mice in the first area and the second area;

[0011] The neuronal calcium signal acquisition system acquires and analyzes calcium signals in a specific brain region of a mouse, and includes a signal analysis control terminal and a calcium signal acquisition device pre-embedded in the specific brain region of the mouse. An interface for connecting the calcium signal acquisition device and the signal analysis control terminal is left on the head of the mouse. The signal analysis control terminal and the calcium signal acquisition device are connected by an optical fiber via the interface, and a calcium ion indicator is also pre-injected in the specific brain region.

[0012] Preferably, the first area delivers odor through a first multi-channel odor delivery system, and the second area delivers odor through a second multi-channel odor delivery system, and the properties, concentrations and delivery speeds of the odors can be controlled separately.

[0013] Preferably, the calcium signal acquisition device is a ceramic pin.

[0014] Preferably, the tail of the ceramic pin is fixed to the surface of the mouse skull using a combination of biological glue and dental cement.

[0015] Preferably, the motion trajectory recording system also records the movement speed of the mouse.

[0016] Preferably, the controllable delivery multi-channel odor conditional preference device includes an outer box and an inner box, the inner box is arranged inside the outer box, the first area, the second area and the transition area are arranged in the inner box, and controllable partitions are arranged between the first area, the second area and the transition area.

[0017] Preferably, the motion trajectory recording system includes a first camera and a second camera, and the first camera and the second camera are arranged on the top outside the inner box body and correspond to the first area and the second area respectively.

[0018] Preferably, the inner box is connected to an air pump.

[0019] The present invention also provides a method for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, using the above-mentioned device for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, comprising the following steps:

[0020] Before collection, the calcium signal collection device of the neuronal calcium signal collection system is embedded in the specific brain area of ​​the mouse in advance, the calcium ion indicator is injected in the specific brain area of ​​the mouse in advance, and the connection port is reserved;

[0021] At the beginning of collection, the mouse is first placed in the transition area of ​​the controllably delivered multi-channel odor conditional preference device, odors of different valences are introduced into the first area and the second area, and the first area and the second area are connected to the transition area by control, and the mouse starts to move freely from the transition area and can freely shuttle between the three areas;

[0022] During the free movement of the mice, the motion trajectory recording system is used to count the movement trajectory and residence time of the mice in the specific odor exposure area, and the specific odor valence of the mice in the first area and the second area is judged to be positive, negative or neutral based on the difference in density of the motion trajectory and the difference in residence time.

[0023] Preferably, a craniotomy is performed on mice at least 3-4 weeks before collection, and the calcium signal acquisition device of the neuronal calcium signal acquisition system is embedded in a specific brain region of the mouse in advance, and the calcium ion indicator is injected in the specific brain region of the mouse in advance.

[0024] Preferably, judging whether the specific odor in the first area and the second area has a positive odor valence or a negative odor valence for the mouse based on the density difference and the residence time difference of the movement trajectory specifically includes:

[0025] If the movement trajectory of the mouse in the first area is denser than that in the second area, and the difference between the residence time in the first area and the residence time in the second area is greater than a first preset threshold, it is determined that the mouse exhibits a positive odor valence to the odor delivered in the first area, and a neutral odor valence or a negative odor valence to the odor delivered in the second area;

[0026] On the contrary, if the movement trajectory of the mouse in the first area is sparser than that in the second area, and the difference between the residence time in the first area and the residence time in the second area is less than or equal to a first preset threshold, it is determined that the mouse exhibits a negative odor valence to the odor delivered in the first area, and a neutral odor valence or a positive odor valence to the odor delivered in the second area.

[0027] Preferably, when the absolute value of the difference in residence time of the mouse in the first area and the second area is greater than a second preset threshold, it is determined that the mouse has undergone a valence shift in response to the delivered odor.

[0028] Preferably, when the mouse starts to move freely, the neuronal calcium signals of the specific brain regions of the mouse are collected by the neuronal calcium signal collection system, and the changes in the neuronal activities of the specific brain regions of the mouse during the valence transition are compared, and the behavioral information of the mouse in response to the valence transitions of different odors and the calcium signal change information of neurons in different brain regions are associated.

[0029] Preferably, the odor valence of a specific odor is also auxiliary judged based on the movement speed of the mouse.

[0030] Preferably, when the movement speed of the mouse decreases from the movement speed of the mouse under natural conditions, the mouse exhibits a positive odor valence to the smell of the current area; when the movement speed of the mouse increases from the movement speed of the mouse under natural conditions, the mouse exhibits a negative odor valence to the smell of the current area.

[0031] Preferably, key nodes where the mouse shuttles between odor areas of different valences are marked, and real-time calcium signal collection is performed in specific brain areas. The key nodes include when the movement speed changes by more than a preset ratio, when the movement direction changes by more than a preset angle within a continuous preset time, and when the mouse is at the boundary between the first area and the second area.

[0032] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0033] The present invention constructs a multi-channel odor conditional preference device for controllable delivery of a mouse odor valence characterization behavior box, and uses an in vivo neuron calcium signal acquisition system to collect and record neuron calcium signals in a specific brain region in real time. The time points of the mouse's switching and shuttling between two side regions into which different odors are introduced are used as time markers, and the mouse's attraction, avoidance or stay behavior is used as a behavioral characterization of the odor valence to associate with the neuron calcium signals in the brain region, thereby improving the accuracy of the association without affecting the mouse's free movement in a limited activity area. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 Schematic diagram of a device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to an embodiment of the present invention;

[0035] Figure 2 1 is a schematic diagram of the movement trajectory of mice in the negative odor valence group in the first area and the second area according to an embodiment of the present invention, wherein the odor delivered to the first area on the left is TMT, an odor that mice are naturally disgusted with, and the second area on the right as a control group delivers air flow;

[0036] Figure 3 1 is a schematic diagram of the movement trajectory of mice in the first and second regions of a positive odor valence group according to an embodiment of the present invention, wherein the odor delivered to the second region on the right is vanillin, an odor that mice innately prefer, and the odor delivered to the first region on the left as a control group is air flow;

[0037] Figure 4 1 is a graph showing the response of neurons in the mouse olfactory cortex during the delivery of different characteristic odors according to an embodiment of the present invention, wherein the odors delivered from top to bottom in the curve are TMT, ethanol, maltol, vanillin, linalool and air flow.

[0038] In the figure, 1-neuronal calcium signal acquisition system; 2-motion trajectory recording system; 3-multi-channel odor conditional preference device with controllable delivery, 4-first multi-channel odor delivery system; 5-second multi-channel odor delivery system; 6-first area; 7-transition area; 8-second area; 9-vacuum pump. DETAILED DESCRIPTION

[0039] According to a specific implementation scheme of the present invention, the technical scheme of the present invention is further described in conjunction with the accompanying drawings.

[0040] The present invention provides a device for comparing changes in neuronal activity in specific brain regions during the process of odor potency change in mice in vivo, comprising a controllable delivery multi-channel odor conditional preference device 3, a motion trajectory recording system 2, and a neuronal calcium signal acquisition system 1; during acquisition, the mouse is placed in the controllable delivery multi-channel odor conditional preference device 3;

[0041] The controllable delivery multi-channel odor condition preference device 3 comprises at least three areas, a first area 6, a second area 8 and a transition area 7; different odors can be introduced into the first area 6 and the second area 8, and the first area 6, the second area 8 and the transition area 7 can be connected through control;

[0042] The movement trajectory recording system 2 records the movement trajectory and residence time of mice in the first area 6 and the second area 8;

[0043] The neuronal calcium signal acquisition system 1 acquires and analyzes calcium signals in a specific brain region of a mouse, and includes a signal analysis control terminal and a calcium signal acquisition device pre-embedded in the specific brain region of the mouse. An interface for connecting the calcium signal acquisition device and the signal analysis control terminal is left on the head of the mouse. The signal analysis control terminal and the calcium signal acquisition device are connected by optical fiber via the interface, and a calcium ion indicator is also pre-injected in the specific brain region.

[0044] According to a specific embodiment of the present invention, the first area 6 delivers odor through a first multi-channel odor delivery system 4, and the second area 8 delivers odor through a second multi-channel odor delivery system 5, and the properties, concentrations and delivery speeds of the odors can be controlled separately.

[0045] According to a specific embodiment of the present invention, the calcium signal acquisition device is a ceramic pin.

[0046] According to a specific embodiment of the present invention, the tail of the ceramic pin is fixed to the surface of the mouse skull using a combination of biological glue and dental cement.

[0047] According to a specific embodiment of the present invention, the motion trajectory recording system 2 also records the movement speed of the mouse.

[0048] According to a specific embodiment of the present invention, the controllable delivery multi-channel odor condition preference device 3 includes an outer box and an inner box, the inner box is arranged inside the outer box, the first area 6, the second area 8 and the transition area 7 are arranged in the inner box, and controllable partitions are arranged between the first area 6, the second area 8 and the transition area 7.

[0049] According to a specific embodiment of the present invention, the motion trajectory recording system includes a first camera and a second camera, and the first camera and the second camera are arranged on the top outside the inner box body and correspond to the first area and the second area respectively.

[0050] According to a specific embodiment of the present invention, the inner box is connected to an air pump 9.

[0051] The present invention also provides a method for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, using the above-mentioned device for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, comprising the following steps:

[0052] Before collecting, the calcium signal collection device of the neuronal calcium signal collection system 1 is embedded in the specific brain area of ​​the mouse in advance, the calcium ion indicator is pre-injected into the specific brain area of ​​the mouse, and the connection port is reserved;

[0053] At the beginning of collection, the mouse is first placed in the transition area of ​​the controllable delivery multi-channel odor condition preference device 3, and odors of different valences are introduced into the first area 6 and the second area 8. The first area 6 and the second area 8 are connected to the transition area 7 through control. The mouse starts to move freely from the transition area 7 and can freely shuttle between the three areas.

[0054] During the free movement of the mouse, the motion trajectory recording system 2 is used to count the movement trajectory and residence time of the mouse in the specific odor exposure area, and the specific odor valence of the mouse in the first area and the second area is judged to be positive, negative or neutral based on the difference in density of the motion trajectory and the difference in residence time.

[0055] According to a specific embodiment of the present invention, a craniotomy is performed on mice at least 3-4 weeks before collection, and the calcium signal collection device of the neuronal calcium signal collection system 1 is embedded in a specific brain region of the mouse in advance, and the calcium ion indicator is pre-injected into the specific brain region of the mouse.

[0056] According to a specific embodiment of the present invention, judging whether the specific odor in the first area 6 and the second area 8 of the mouse has a positive odor valence or a negative odor valence according to the density difference and the residence time difference of the movement trajectory specifically includes:

[0057] If the movement trajectory of the mouse in the first area 6 is denser than that in the second area 8, and the difference between the residence time in the first area 6 and the residence time in the second area 8 is greater than a first preset threshold, it is determined that the mouse exhibits a positive odor valence to the odor delivered in the first area 6, and a neutral odor valence or a negative odor valence to the odor delivered in the second area 8;

[0058] On the contrary, if the movement trajectory of the mouse in the first area 6 is sparser than that in the second area 8, and the difference between the residence time in the first area 6 and the residence time in the second area 8 is less than or equal to a first preset threshold, it is determined that the mouse exhibits a negative odor valence to the odor delivered in the first area 6, and a neutral odor valence or a positive odor valence to the odor delivered in the second area 8.

[0059] According to a specific embodiment of the present invention, when the absolute value of the difference in residence time of the mouse in the first area 6 and the second area 8 is greater than a second preset threshold, it is determined that the mouse has undergone a valence shift in response to the delivered odor.

[0060] According to a specific embodiment of the present invention, when a mouse starts to move freely, the neuronal calcium signals of a specific brain region of the mouse are collected by the neuronal calcium signal collection system 1, and the changes in the neuronal activities of the specific brain regions of the mouse during the valence transition are compared, and the behavioral information of the mouse to the changes in the valence of different odors and the calcium signal change information of neurons in different brain regions are associated.

[0061] According to a specific embodiment of the present invention, the odor valence of a specific odor is also auxiliary judged based on the movement speed of the mouse.

[0062] According to a specific embodiment of the present invention, when the movement speed of the mouse decreases from the movement speed of the mouse under natural conditions, the mouse exhibits a positive odor valence to the odor of the current area; when the movement speed of the mouse increases from the movement speed of the mouse under natural conditions, the mouse exhibits a negative odor valence to the odor of the current area.

[0063] According to a specific embodiment of the present invention, key nodes of the mouse passing through odor areas of different valences are marked, and real-time calcium signal collection of specific brain areas is performed, and the key nodes include when the movement speed changes by more than a preset ratio, when the movement direction changes by more than a preset angle within a continuous preset time, and when the mouse is at the boundary between the first area and the second area.

[0064] Example 1

[0065] The following is a detailed description of a device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice according to a specific embodiment of the present invention.

[0066] The present invention provides a device for comparing changes in neuronal activity in specific brain regions during the process of odor potency change in mice in vivo, comprising a controllable delivery multi-channel odor conditional preference device 3, a motion trajectory recording system 2, and a neuronal calcium signal acquisition system 1; during acquisition, the mouse is placed in the controllable delivery multi-channel odor conditional preference device 3;

[0067] The controllable delivery multi-channel odor condition preference device 3 comprises at least three areas, a first area 6, a second area 8 and a transition area 7; different odors can be introduced into the first area 6 and the second area 8, and the first area 6, the second area 8 and the transition area 7 can be connected through control;

[0068] The movement trajectory recording system 2 records the movement trajectory and residence time of mice in the first area 6 and the second area 8;

[0069] The neuronal calcium signal acquisition system 1 acquires and analyzes calcium signals in a specific brain region of a mouse, and includes a signal analysis control terminal and a calcium signal acquisition device pre-embedded in the specific brain region of the mouse. An interface for connecting the calcium signal acquisition device and the signal analysis control terminal is left on the head of the mouse. The signal analysis control terminal and the calcium signal acquisition device are connected by optical fiber via the interface, and a calcium ion indicator is also pre-injected in the specific brain region.

[0070] Example 2

[0071] The following is a detailed description of a device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice according to a specific embodiment of the present invention.

[0072] The present invention provides a device for comparing changes in neuronal activity in specific brain regions during the process of odor potency change in mice in vivo, comprising a controllable delivery multi-channel odor conditional preference device 3, a motion trajectory recording system 2, and a neuronal calcium signal acquisition system 1; during acquisition, the mouse is placed in the controllable delivery multi-channel odor conditional preference device 3;

[0073] The controllable delivery multi-channel odor condition preference device 3 comprises at least three areas, a first area 6, a second area 8 and a transition area 7; different odors can be introduced into the first area 6 and the second area 8, and the first area 6, the second area 8 and the transition area 7 can be connected through control;

[0074] The movement trajectory recording system 2 records the movement trajectory and residence time of mice in the first area 6 and the second area 8;

[0075] The neuronal calcium signal acquisition system 1 acquires and analyzes calcium signals in a specific brain region of a mouse, and includes a signal analysis control terminal and a calcium signal acquisition device pre-buried in the specific brain region of the mouse. An interface for connecting the calcium signal acquisition device and the signal analysis control terminal is left on the head of the mouse, and the signal analysis control terminal and the calcium signal acquisition device are connected by an optical fiber via the interface. A calcium ion indicator is also pre-injected into the specific brain region. The calcium signal acquisition device is a ceramic pin, and the tail of the ceramic pin is fixed to the surface of the mouse skull using a combination of biological glue and dental cement.

[0076] The first area 6 delivers odor through a first multi-channel odor delivery system 4, and the second area 8 delivers odor through a second multi-channel odor delivery system 5, and the properties, concentration and delivery speed of the odors can be controlled separately.

[0077] The motion track recording system 2 also records the movement speed of the mouse.

[0078] Among them, the controllable delivery multi-channel odor condition preference device 3 includes an outer box and an inner box, the inner box is arranged inside the outer box, the first area 6, the second area 8 and the transition area 7 are arranged in the inner box, and controllable partitions are arranged between the first area 6, the second area 8 and the transition area 7.

[0079] The motion trajectory recording system includes a first camera and a second camera, wherein the first camera and the second camera are arranged on the top outside the inner box body and correspond to the first area and the second area respectively.

[0080] Wherein, the inner box is connected to an air pump 9.

[0081] Example 3

[0082] The following is a detailed description of the method of the present invention for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo.

[0083] The present invention also provides a method for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, using the above-mentioned device for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, comprising the following steps:

[0084] Before collection, the calcium signal collection device of the neuronal calcium signal collection system 1 is pre-embedded in a specific brain region of the mouse, the calcium ion indicator is pre-injected into the specific brain region of the mouse, and the connection port is reserved;

[0085] At the beginning of collection, the mouse is first placed in the transition area of ​​the controllable delivery multi-channel odor condition preference device 3, and odors of different valences are introduced into the first area 6 and the second area 8. The first area 6 and the second area 8 are connected to the transition area 7 through control. The mouse starts to move freely from the transition area 7 and can freely shuttle between the three areas.

[0086] During the free movement of the mouse, the motion trajectory recording system 2 is used to count the movement trajectory and residence time of the mouse in the specific odor exposure area, and the specific odor valence of the mouse in the first area and the second area is judged to be positive, negative or neutral based on the difference in density of the motion trajectory and the difference in residence time.

[0087] Example 4

[0088] The following is a detailed description of the method of the present invention for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo.

[0089] The present invention also provides a method for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, using the above-mentioned device for comparing changes in neuronal activity in specific brain regions during the process of odor valence change in mice in vivo, comprising the following steps:

[0090] Perform a craniotomy on mice at least 3-4 weeks before collection, embed the calcium signal collection device of the neuronal calcium signal collection system 1 in the specific brain region of the mouse in advance, inject the calcium ion indicator in the specific brain region of the mouse in advance, and retain the connection port;

[0091] At the beginning of collection, the mouse is first placed in the transition area of ​​the controllable delivery multi-channel odor condition preference device 3, and odors of different valences are introduced into the first area 6 and the second area 8. The first area 6 and the second area 8 are connected to the transition area 7 through control. The mouse starts to move freely from the transition area 7 and can freely shuttle between the three areas.

[0092] During the free movement of the mouse, the motion trajectory recording system 2 is used to count the movement trajectory and residence time of the mouse in the specific odor exposure area, and the specific odor valence of the mouse in the first area and the second area is judged to be positive, negative or neutral based on the difference in density of the motion trajectory and the difference in residence time.

[0093] Wherein, judging whether the specific odors in the first area 6 and the second area 8 of the mouse have positive odor valence or negative odor valence according to the density difference and the residence time difference of the movement trajectory specifically includes:

[0094] If the movement trajectory of the mouse in the first area 6 is denser than that in the second area 8, and the difference between the residence time in the first area 6 and the residence time in the second area 8 is greater than a first preset threshold (the setting of the first preset threshold ensures that the residence time difference is significantly large, that is, the statistical difference p value of the two residence times is ensured to be ≤0.05), it is determined that the mouse exhibits a positive odor valence to the odor delivered in the first area 6, and a neutral odor valence or a negative odor valence to the odor delivered in the second area 8;

[0095] On the contrary, if the movement trajectory of the mouse in the first area 6 is sparser than that in the second area 8, and the difference between the residence time in the first area 6 and the residence time in the second area 8 is less than or equal to a first preset threshold, it is determined that the mouse exhibits a negative odor valence to the odor delivered in the first area 6, and a neutral odor valence or a positive odor valence to the odor delivered in the second area 8.

[0096] When the absolute value of the difference in residence time of the mouse in the first area 6 and the second area 8 is greater than a second preset threshold, it is determined that the mouse has undergone a valence change in response to the delivered odor.

[0097] Among them, when the mouse starts to move freely, the neuronal calcium signals of the specific brain regions of the mouse are collected by the neuronal calcium signal collection system 1, and the changes in the neuronal activities of the specific brain regions of the mouse during the valence change process are compared, and the behavioral information of the mouse to the changes in different odor valences and the calcium signal change information of neurons in different brain regions are associated.

[0098] The odor valence of a specific odor is also assisted in the judgment based on the movement speed of the mouse.

[0099] Among them, when the movement speed of the mouse is reduced from the movement speed of the mouse under natural conditions, the mouse shows a positive odor valence to the smell of the current area; when the movement speed of the mouse is increased from the movement speed of the mouse under natural conditions, the mouse shows a negative odor valence to the smell of the current area. If the bilateral movement speed of the mouse is significantly lower or higher than the movement speed of the mouse under natural conditions, such as 10m / min, and the bilateral residence time has a significant difference, a judgment can be made, and the movement speed cannot be used as the only judgment indicator.

[0100] Among them, key nodes when the mouse shuttles between odor areas of different valences are marked, and real-time calcium signals from specific brain areas are collected. The key nodes include when the movement speed changes by more than a preset ratio, when the movement direction changes by more than a preset angle within a continuous preset time, and when the mouse is at the boundary between the first area and the second area.

[0101] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention are included in the protection scope of the present invention.

Claims

1. A device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo, characterized in that: It includes a multi-channel odor conditional preference device with controllable delivery, a motion trajectory recording system and a neuron calcium signal acquisition system; during acquisition, a mouse is placed in the multi-channel odor conditional preference device with controllable delivery; The controllable delivery multi-channel odor condition preference device comprises at least three areas, a first area, a second area and a transition area, wherein the transition area is located between the first area and the second area; different odors can be introduced into the first area and the second area, and the first area, the second area and the transition area can be connected through control; The motion trajectory recording system records the motion trajectory and residence time of mice in the first area and the second area; The neuronal calcium signal acquisition system acquires and analyzes calcium signals in a specific brain region of a mouse, and includes a signal analysis control terminal and a calcium signal acquisition device pre-embedded in the specific brain region of the mouse. An interface for connecting the calcium signal acquisition device and the signal analysis control terminal is left on the head of the mouse. The signal analysis control terminal and the calcium signal acquisition device are connected by an optical fiber via the interface, and a calcium ion indicator is also pre-injected in the specific brain region.

2. The device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to claim 1, characterized in that: The first area delivers the scent through a first multi-channel scent delivery system, and the second area delivers the scent through a second multi-channel scent delivery system, and the properties, concentrations and delivery speeds of the scents can be controlled separately.

3. The device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to claim 1, characterized in that: The calcium signal acquisition device is a ceramic pin.

4. The device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to claim 3, characterized in that: The tail of the ceramic pin is fixed on the surface of the mouse skull using a combination of biological glue and dental cement.

5. The device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to claim 1, characterized in that: The movement track recording system also records the movement speed of the mouse.

6. The device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to claim 1, characterized in that: The controllable delivery multi-channel odor conditional preference device includes an outer box and an inner box, the inner box is arranged inside the outer box, the first area, the second area and the transition area are arranged in the inner box, and controllable partitions are arranged between the first area, the second area and the transition area.

7. The device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to claim 6, characterized in that: The motion track recording system includes a first camera and a second camera, wherein the first camera and the second camera are arranged on the top outside the inner box body and correspond to the first area and the second area respectively.

8. The device for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo according to claim 6, characterized in that: The inner box is connected with an air pump.

9. A method for comparing changes in neuronal activity in specific brain regions during odor valence changes in mice in vivo, characterized in that: The device for comparing changes in neuronal activity in specific brain regions of mice during odor valence transformation in vivo according to any one of claims 1 to 8 comprises the following steps: At the beginning of collection, the mouse is first placed in the transition area of ​​the controllably delivered multi-channel odor conditional preference device, odors of different valences are introduced into the first area and the second area, and the first area and the second area are connected to the transition area by control, and the mouse starts to move freely from the transition area and can freely shuttle between the three areas; During the free movement of the mice, the motion trajectory recording system is used to count the movement trajectory and residence time of the mice in the specific odor exposure area, and the specific odor valence of the mice in the first area and the second area is judged to be positive, negative or neutral based on the difference in density of the motion trajectory and the difference in residence time.

10. The method for in vivo comparison of changes in neuronal activity in specific brain regions during odor valence changes in mice according to claim 9, characterized in that: Judging whether the specific odors in the first area and the second area have a positive odor valence or a negative odor valence for the mouse according to the density difference and the residence time difference of the movement tracks specifically includes: If the movement trajectory of the mouse in the first area is denser than that in the second area, and the difference between the residence time in the first area and the residence time in the second area is greater than a first preset threshold, it is determined that the mouse exhibits a positive odor valence to the odor delivered in the first area, and a neutral odor valence or a negative odor valence to the odor delivered in the second area; On the contrary, if the movement trajectory of the mouse in the first area is sparser than that in the second area, and the difference between the residence time in the first area and the residence time in the second area is less than or equal to a first preset threshold, it is determined that the mouse exhibits a negative odor valence to the odor delivered in the first area, and a neutral odor valence or a positive odor valence to the odor delivered in the second area.

11. The method for in vivo comparison of changes in neuronal activity in specific brain regions during odor valence changes in mice according to claim 10, characterized in that: When the absolute value of the difference in residence time of the mouse in the first area and the second area is greater than a second preset threshold, it is determined that the mouse has undergone a valence shift in response to the delivered odor.

12. The method for in vivo comparison of changes in neuronal activity in specific brain regions during odor valence changes in mice according to claim 11, characterized in that: When the mouse starts to move freely, the neuronal calcium signals in the specific brain regions of the mouse are collected by the neuronal calcium signal collection system, and the changes in the neuronal activities in the specific brain regions of the mouse during the valence transition are compared, and the behavioral information of the mouse in response to the changes in odor valence and the calcium signal change information of neurons in different brain regions are associated.

13. The method for in vivo comparison of changes in neuronal activity in specific brain regions during odor valence changes in mice according to claim 9, characterized in that: The movement speed of the mouse is also used to assist in determining the odor valence of a specific odor.

14. The method for in vivo comparison of changes in neuronal activity in specific brain regions during odor valence changes in mice according to claim 13, characterized in that: When the movement speed of the mouse decreases from the movement speed of the mouse under natural conditions, the mouse expresses a positive odor valence to the smell of the current area; when the movement speed of the mouse increases from the movement speed of the mouse under natural conditions, the mouse expresses a negative odor valence to the smell of the current area.

15. The method for in vivo comparison of changes in neuronal activity in specific brain regions during odor valence changes in mice according to claim 14, characterized in that: The key nodes when the mouse shuttles between odor areas of different valences are marked, and real-time calcium signals from specific brain areas are collected. The key nodes include when the movement speed changes by more than a preset ratio, when the movement direction changes by more than a preset angle within a continuous preset time, and when the mouse is at the boundary between the first area and the second area.

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