Animal experiment model and method for calcium signal recording

Abstract

The invention relates to the technical field of medical experiment animal models, in particular to an animal experiment model and method for calcium signal recording, and the model comprises the following steps: introducing a fluorescent marker from a specific area of a selected target body into neurons of the target body, and arranging a microscopic camera fixing seat at the specific area of the target body, and observing the brain activity of the target body by using a microscope camera. According to the invention, intravenous administration and calcium signal recording model animal construction is realized, and subsequent research on drug effects and treatment mechanisms is facilitated by observing the change of calcium signals at the moment of drug intravenous blood vessel infusion.

Description

Technical Field

[0001] The present invention relates to the technical field of medical experimental animal models, and more particularly, to an animal experimental model and method for calcium signal recording. Background Art

[0002] In the study of the nervous system, microscopic imaging of calcium signals provides unique insights into neuronal activity (the activity of nerve cells) and neural network function (the interaction between nerve cells: for example, whether neuron A can control neuron B, or whether neuron B can control neuron A). Microscopic calcium signal imaging (recording the calcium signal activity of nerve cells using a micro camera) can dynamically observe nerve activity. The change in calcium ion concentration in nerve cells is closely related to cell activity. By recording the change in calcium signals, it is possible to understand whether the response of nerve cells to certain events is excitatory or inhibitory, and how they interact in the neural network. The following benefits can be obtained through microscopic imaging: 1. Understanding the relationship between neural network connectivity and behavioral performance: By recording calcium signals (the activity signals of calcium ions), researchers can study the activity patterns of different neuron populations in the neural network, and thus understand the relationship between these patterns and the behavioral performance of animals. 2. Exploring the learning and memory processes: Calcium signal imaging helps to study the changes in neuronal activity during the formation of learning and memory. By tracking the activity of specific neurons, its role in the formation of learning and memory can be revealed. 3. Revealing diseases and nervous system abnormalities: By observing abnormal patterns of neuronal activity (the changes in the activity of nerve cells in the diseased state), it helps to understand the disease mechanism and may provide new directions for disease treatment. 4. Verifying neural models and theories: Calcium signal imaging data can be used to verify neural network models and theories. Through the acquisition of experimental data, scientists can verify hypotheses in neuroscience and promote a deeper understanding of the function of the nervous system.

[0003] However, one of the most common and unresolved problems in current microscopic imaging is that it can only record the calcium signals of freely moving animals, and it is impossible to synchronously record and analyze the changes in the calcium signal responses of nerve cells in animals at the moment of intravenous drug administration (rapid increase in blood drug concentration) in scientific research, that is, it is impossible to record the changes in the state of nerve cells in the brain region of freely moving animals at the moment of receiving intravenous drug delivery and during drug treatment.

[0004] Therefore, the existing technology still has deficiencies and needs to be improved. Summary of the Invention

[0005] The embodiments of the present invention provide an animal experimental model and method for calcium signal recording, so as to achieve the purpose of dynamically recording and analyzing the changes in the calcium signals of animal neurons before and after intravenous drug delivery.

[0006] According to an embodiment of the present invention, an animal experiment model for calcium signal recording is provided, including the following steps:

[0007] Introduce a preset fluorescent marker into the neurons of the target body from a specific area of the selected target body;

[0008] Implant the autofocus module into the specific area;

[0009] Set up a microscope camera mount in the specific area;

[0010] Set up a microscope camera on the microscope camera mount for observing the specific area of the target body;

[0011] Set one end of the drug delivery device to be connected to the jugular vein blood vessel of the target body, and the other end to be connected to the computer module. The computer module controls the drug delivery device to administer drugs to the target body and obtains the data observed by the microscope camera.

[0012] In one embodiment, the autofocus module is a glass autofocus prism.

[0013] In one embodiment, the autofocus module is fixed by photocuring resin.

[0014] In one embodiment, the model further includes:

[0015] The camera data acquisition module, with one end connected to the microscope camera and the other end connected to the computer module, is used to record the data of the specific area and transmit it to the computer module.

[0016] In one embodiment, the model further includes:

[0017] The nose touch panel, connected to the computer module, is used to collect the nose touch signal of the target body and transmit a synchronization signal to the computer module.

[0018] In one embodiment, the model further includes:

[0019] The nose touch data acquisition module, with one end connected to the nose touch panel and the other end connected to the computer module, and the nose touch panel is connected to the computer module through a data acquisition card to send a synchronization signal.

[0020] An animal experiment method for calcium signal recording, including the application of the animal experiment model for calcium signal recording according to any one of the above, the method includes:

[0021] Based on the drug administration signal, the computer module controls the drug delivery device to administer drugs to the target body;

[0022] Use the microscope camera to observe the specific area of the target body.

[0023] In one embodiment, after the computer module controls the administration device to administer medicine to the target based on the administration signal, the following steps are further included:

[0024] Collect data of a specific area through the camera data acquisition module and transmit it to the computer module.

[0025] In one embodiment, after observing a specific area of the target using a microscopic camera, the following steps are further included:

[0026] Real-time collect the nasal touch signal of the target through the nasal touch module;

[0027] Based on the nasal touch signal, the data acquisition module sends a synchronization signal to the computer module;

[0028] Based on the synchronization signal, the computer generates a synchronization file, and the synchronization file includes the video data obtained by the microscopic camera.

[0029] In one embodiment, after real-time collecting the nasal touch signal of the target through the nasal touch module, the following steps are further included:

[0030] Based on the nasal touch signal, the nasal touch data acquisition module drives the administration device to administer medicine to the target.

[0031] In the animal experiment model and method for calcium signal recording in the embodiments of the present invention, a fluorescent marker is introduced into the neurons of the target from a specific area of the selected target. A microscopic camera fixing base is set at the specific area of the target, and the brain activity of the target is observed using a microscopic camera. The present invention realizes the construction of an intravenous administration and microscopic calcium signal recording model animal, and by observing the changes in the calcium signal of the animal at the moment of intravenous infusion of the drug into the blood vessel, it is convenient to explore the drug effect and treatment mechanism subsequently. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The drawings described herein are used to provide a further understanding of the present invention, form a part of this application, and the schematic embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation to the present invention. In the drawings:

[0033] Figure 1 is a module diagram of the animal experiment model for calcium signal recording of the present invention;

[0034] Figure 2 is a schematic principle diagram of the animal experiment model for calcium signal recording of the present invention;

[0035] Figure 3 is a schematic diagram of the target of the present invention;

[0036] Figure 4 is a schematic diagram of intravenous injection of the present invention;

[0037] Figure 5This is a flowchart of the animal experiment method for calcium signal recording in the present invention.

[0038] Reference numerals: 1 - target body, 2 - camera data acquisition module, 3 - computer module, 4 - nose touch panel, 5 - drug delivery device, 6 - nose touch data acquisition module, 7 - drug delivery catheter, 8 - microscope camera fixing base, 9 - autofocus module. Detailed implementation manners

[0039] In order to enable those skilled in the art to better understand the solution of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0040] It should be noted that the terms "first", "second", etc. in the description and claims of the present invention and the above drawings are used to distinguish similar objects, and do not necessarily need to describe a specific order or sequence. It should be understood that such data can be interchanged under appropriate circumstances so that the embodiments of the present invention described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "comprising" and "having" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product or device including a series of steps or units does not necessarily have to be limited to those steps or units clearly listed, but may include other steps or units not clearly listed or inherent to these processes, methods, products or devices.

[0041] Embodiment 1

[0042] According to an embodiment of the present invention, an animal experiment model for calcium signal recording is provided. Refer to Figures 1 to 4 , the model includes:

[0043] Introduce a preset fluorescent marker into the neurons of the target body 1 from a specific area of the selected target body 1;

[0044] Implant the autofocus module 9 into the specific area;

[0045] Set the microscope camera fixing base 8 in the specific area;

[0046] Set a microscope camera on the microscope camera fixing base 8 for observing the specific area of the target body 1;

[0047] One end of the drug delivery device 5 is connected to the jugular vein blood vessel of the target body 1, and the other end is connected to the computer module 3. The computer module 3 is used to control the drug delivery device 5 to administer drugs to the target body 1 and obtain the data observed by the microscope camera.

[0048] In the present invention, a fluorescent marker is introduced into the neurons of the target body 1 from a specific region of the selected target body 1. A microscope camera fixing base 8 is arranged at the specific region of the target body 1, and the microscope camera is used to observe the brain activity of the target body 1. The present invention realizes the construction of an intravenous drug administration and microscopic calcium signal recording model animal. By observing the changes in the calcium signal of the animal at the moment of intravenous blood vessel infusion of the drug, it is convenient to explore the drug action and treatment mechanism subsequently.

[0049] Specifically, the target body 1 can be an experimental mouse. The selected fluorescent marker or gene can be introduced into mouse neurons through gene transduction, which can be achieved through a viral vector (such as AAV) + transgenic mouse to ensure sufficient expression in the target neurons. Operating steps: Anesthetize the mouse with an anesthetic drug, remove the hair on the head, cut open the scalp, level the mouse's head, drill a hole in the skull surface of the specific brain region with a skull drill, and inject a virus with a specific fluorescent label into the specific brain region of the mouse. Among them, the fluorescent marker can be selected as calcium ion.

[0050] In one embodiment, referring to Figure 1 and Figure 3 , the autofocus module 9 is a glass autofocus prism. Specifically, the glass autofocus prism (Lens) for microscopic imaging is implanted along the virus injection channel into the specific brain region (the selected specific region) of the mouse, and the Lens is fixed to the head with a light-curing resin. The focusing plane of the Lens is adjusted through the microscope camera, and a microscope camera fixing base 8 is installed on the mouse's head. During the experiment, the microscope camera is connected to the microscope camera fixing base 8.

[0051] Anesthetize the mouse with an anesthetic drug, remove the hair on the right neck skin of the mouse with a hair removal cream, disinfect and then cut open the skin to expose the external jugular vein blood vessel. Referring to Figure 4 , implant a silicone tube into the blood vessel, and the other end of the silicone tube is connected to the drug delivery device 5 through a back incision. Wait for the mouse to wake up to complete the preliminary construction of the model animal.

[0052] In one embodiment, referring to Figure 1 and Figure 2 , the model further includes:

[0053] The camera data acquisition module 2, one end is connected to the microscope camera, and the other end is connected to the computer module 3, which is used to record the data of the specific region and transmit it to the computer module 3.

[0054] During the experiment, a specific area of the mouse is observed through a microscope camera, and the camera data acquisition module 2 obtains the data observed by the microscope camera, and then transmits the recorded specific area data to the computer module 3, and the computer module 3 processes and analyzes the nerve cells in the specific area.

[0055] In one embodiment, referring to Figure 1 and Figure 2 , the model further includes:

[0056] A nose touch panel 4, connected to the computer module 3, for collecting the nose touch signal of the target body 1 and transmitting a synchronization signal to the computer module 3.

[0057] When the mouse touches the nose touch panel 4, a nose touch signal will be generated, and then the signal will be transmitted to the computer module 3. The computer module 3 controls the drug delivery device 5 to administer drugs to the mouse according to the signal.

[0058] In one embodiment, referring to Figure 1 and Figure 2 , the model further includes:

[0059] A nose touch data acquisition module 6, one end is connected to the nose touch panel 4, and the other end is connected to the computer module 3. The nose touch panel 4 is connected to the computer module 3 by sending a synchronization signal through a data acquisition card.

[0060] Example 2

[0061] According to another embodiment of the present invention, an animal experiment method for calcium signal recording is provided. Refer to Figure 1 , Figure 2 and Figure 5 , including:

[0062] S100: Based on the drug delivery signal, the computer module 3 controls the drug delivery device 5 to administer drugs to the target body 1;

[0063] S200: Use a microscope camera to observe a specific area of the target body 1.

[0064] After receiving the drug delivery instruction, the computer module 3 outputs a drug delivery signal. The drug delivery device 5 receives the drug delivery signal and delivers the drug to the mouse's intravenous blood vessel through a drug delivery catheter. The microscope camera also receives the drug delivery signal and records it to achieve subsequent synchronous analysis of the data.

[0065] In one embodiment, referring to Figure 1 and Figure 2 , after the computer module 3 controls the drug delivery device 5 to administer drugs to the target body 1 based on the drug delivery signal, it further includes:

[0066] Collect specific area data through the camera data acquisition module 2 and transmit it to the computer module 3.

[0067] In this application, a microscope camera is used to observe the brain activity of the target body 1, realizing the construction of a model animal for intravenous drug administration and microscopic calcium signal recording. The change of the calcium signal of the animal at the moment of intravenous blood vessel infusion of the drug is observed through the microscope camera, and the observed signal is sent to the computer module 3 for analysis, facilitating the subsequent exploration of the drug effect and treatment mechanism.

[0068] In one embodiment, referring to Figure 1 and Figure 2 , after observing a specific area of the target body 1 using the microscope camera, it further includes:

[0069] The nasal touch signal of the target body 1 is collected in real time through the nasal touch module;

[0070] Based on the nasal touch signal, the data acquisition module sends a synchronization signal to the computer module 3;

[0071] Based on the synchronization signal, the computer module 3 generates a synchronization file, and the synchronization file includes the video data obtained by the microscope camera.

[0072] Specifically, during the experiment, the mouse can touch the nasal touch panel 4. Based on the touch of the mouse, the nasal touch panel 4 will transmit the nasal touch signal to the nasal touch data acquisition module 6, and the nasal touch data acquisition module 6 will transmit it to the computer module 3 according to the received nasal touch surface signal. At the same time, it receives the drug administration signal driven by the computer module 3 and sends it to the drug administration device 5. During this process, the computer module 3 will store the information obtained by the microscope camera as a file.

[0073] In one embodiment, referring to Figure 1 and Figure 2 , after collecting the nasal touch signal of the target body 1 in real time through the nasal touch module, it further includes:

[0074] Based on the nasal touch signal, the data acquisition module drives the drug administration device 5 to administer the drug to the target body 1.

[0075] The computer module 3 will obtain the nasal touch data from the nasal touch data acquisition module 6. The nasal touch data includes the time point and the number of times. Then, according to the nasal touch data, it controls the drug administration device 5 to administer the drug, realizing that the drug administration device 5 administers the drug to the mouse based on the touch of the mouse, and then repeats the observation of the mouse through the microscope camera.

[0076] Referring to Figure 1 and Figure 2 , the model principle of this application and the method of applying the model are as follows:

[0077] Model mouse (i.e., target body 1): A microscopic camera mount 8 is provided on the mouse's head. During the experiment, by setting the microscopic camera on the microscopic camera mount 8, the connection between the mouse's head and the microscopic camera is achieved, and it is connected to the drug delivery device 5 through the white drug delivery catheter on the mouse's back.

[0078] Video data acquisition module 2: Connected to the microscopic camera and the computer module 3, and used to transmit the data obtained by the micro camera to the computer module 3.

[0079] Computer module 3: Used to record the video information obtained by the microscopic camera on the mouse's head, record the data signal when the mouse touches the nose touch panel 4, and drive the drug delivery device 5 to deliver drugs to the mouse through the drug delivery catheter on the mouse's back according to this data signal.

[0080] Nose touch panel 4: When the mouse touches the nose touch panel 4, the nose touch panel 4 sends a synchronization signal to the computer module 3 through the nose touch data acquisition module 6, and this synchronization signal can enable the computer module 3 to generate a synchronization document for the video signal of the microscopic camera.

[0081] Drug delivery device 5: Adopts a micro peristaltic pump, and after receiving the drug delivery signal from the nose touch data acquisition module 6, delivers drugs to the model mouse through the drug delivery catheter.

[0082] Nose touch data acquisition module 6: Used to connect the drug delivery device 5 and the nose touch panel 4 to the computer module 3. When receiving the nose touch signal from the nose touch panel 4, it transmits this nose touch signal to the computer module 3, and at the same time receives the feedback drive drug delivery signal from the computer module 3 and sends it to the drug delivery device 5. After the drug delivery device 5 receives the feedback drug delivery signal, it delivers drugs to the mouse.

[0083] In this application, a suitable calcium ion is selected as the indicator, and then the calcium ion indicator is transported to a specific area of the mouse's brain. The calcium ion indicator is introduced into the target brain area by means of virus infection, so that the activities of the infected nerve cells (when the animal is stimulated by certain events, some neurons may be excited, some neurons may be inhibited, and some neurons may not respond to the stimulus) can be observed under a fluorescence microscope (microscopic camera), and captured and recorded by the microscopic camera. The changes in calcium signals are collected, the results are recorded, and the recorded data is analyzed.

[0084] The purpose of the present invention is to construct an experimental animal model combining intravenous catheterization and microscopic imaging, so as to achieve the purpose of dynamically recording the changes in calcium signals of animal neurons before and after intravenous drug delivery. Using this animal model, a large number of drug treatment mechanisms related to brain science can be carried out, and it can be used for subsequent research on the treatment of anesthetic drugs, drugs for neurological diseases (such as Alzheimer's disease, epilepsy, etc.).

[0085] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. An animal experimental model for calcium signal recording, characterized in that, The experimental model includes: Introducing a preset fluorescent marker into the neurons of the target body from a specific region of the selected target body; Implanting an autofocus module into the specific region; Setting a microscope camera mount in the specific region; Setting a microscope camera on the microscope camera mount for observing the specific region of the target body; Setting one end of a drug delivery device to be connected to the jugular vein blood vessel of the target body and the other end to be connected to a computer module, controlling the drug delivery device to administer drugs to the target body through the computer module, and acquiring the data observed by the microscope camera.

2. The animal experimental model for calcium signal recording according to claim 1, characterized in that, The autofocus module is a glass autofocus prism.

3. The animal experimental model for calcium signal recording according to claim 1, characterized in that, Fixing the autofocus module by photocuring resin.

4. The animal experimental model for calcium signal recording according to claim 3, characterized in that, The model further includes: A camera data acquisition module, connected to the microscope camera at one end and to the computer module at the other end, for recording the data of the specific region and transmitting it to the computer module.

5. The animal experimental model for calcium signal recording according to claim 4, characterized in that, The model further includes: A nose touch panel, connected to the computer module, for collecting the nose storage signal of the target body and transmitting a synchronization signal to the computer module.

6. The animal experimental model for calcium signal recording according to claim 5, characterized in that, The model further includes: A nose touch data acquisition module, connected to the nose touch panel at one end and to the computer module at the other end, and the nose touch panel is connected to the computer module through the data acquisition card to send the synchronization signal.

7. An animal experimental method for calcium signal recording, characterized in that, Including the application of the animal experimental model for calcium signal recording according to any one of claims 1-6, the method includes: Based on the drug administration signal, the computer module controls the drug delivery device to administer drugs to the target body; Using a microscope camera to observe the specific region of the target body.

8. The animal experimental method for calcium signal recording according to claim 7, characterized in that, After the computer module controls the drug delivery device to administer drugs to the target body based on the drug administration signal, it further includes: Collecting the data of the specific region through the camera data acquisition module and transmitting it to the computer module.

9. The animal experimental method for calcium signal recording according to claim 8, characterized in that, After using the microscope camera to observe the specific region of the target body, it further includes: Real-time collecting the nose touch signal of the target body through the nose touch module; The data acquisition module sends a synchronization signal to the computer module based on the nose touch signal; Based on the synchronization signal, the computer generates a synchronization file, and the synchronization file includes the video data obtained by the microscope camera.

10. The animal experimental method for calcium signal recording according to claim 8, characterized in that, After real-time collecting the nose touch signal of the target body through the nose touch module, it further includes: The nose touch data acquisition module drives the drug delivery device to administer drugs to the target body based on the nose touch signal.

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