Animal experiment model and method for calcium signal recording

Abstract

The present invention relates to the field of medical experimental animal model technology, and in particular, to an animal experiment model and method for calcium signal recording. The model comprises: introducing a fluorescent marker into neurons of a target subject from a selected specific area of the target subject, arranging a microscope camera fixing seat at the specific area of the target subject, and using a microscope camera to observe the brain activity of the target subject. The present invention achieves the construction of the animal model for intravenous administration and calcium signal recording, and the drug action and treatment mechanisms can be conveniently explored by observing the change of the calcium signal at the instant of intravenous infusion of the drug.

Description

An animal experimental model and method for calcium signal recording Technical Field

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

[0002] In neurological research, microscopic calcium signaling provides unique insights into neuronal activity (the activity of nerve cells) and neural network function (interactions between nerve cells: for example, whether neuron A controls neuron B, or vice versa). Microscopic calcium signaling (using a microscopic camera to record neuron calcium signaling activity) allows for dynamic observation of neural activity. Changes in the calcium ion concentration of nerve cells are closely related to cellular activity. By recording changes in calcium signals, it is possible to understand whether nerve cells respond to certain events with excitatory or inhibitory responses, as well as how they interact within neural networks. Microscopic imaging offers the following benefits: 1. Understanding the relationship between neural network connectivity and behavioral performance: By recording calcium signals (calcium ion activity signals), researchers can study the activity patterns of different neuronal populations in a neural network and understand the relationship between these patterns and animal behavior. 2. Exploring learning and memory processes: Calcium signaling imaging facilitates the study of changes in neuronal activity during learning and memory formation. By tracking the activity of specific neurons, their role in learning and memory formation can be revealed. 3. Revealing Diseases and Nervous System Abnormalities: Observing abnormal patterns of neuronal activity (changes in nerve cell activity under disease conditions) can help us understand disease mechanisms and potentially provide new approaches for disease treatment. 4. Validating Neural Models and Theories: Calcium signal imaging data can be used to validate neural network models and theories. By obtaining experimental data, scientists can verify hypotheses in neuroscience and advance a deeper understanding of nervous system function.

[0003] However, one of the most common and poorly resolved problems with current microscopic imaging is that it can only record calcium signals in freely moving animals. It is unable to synchronously record and analyze changes in animal calcium signal nerve cell responses at the moment of intravenous drug administration (rapid increase in blood drug concentration) in scientific research. In other words, it is impossible to record changes in the state of nerve cells in the brain of freely moving animals at the moment of receiving intravenous drug delivery and when receiving drug treatment.

[0004] Therefore, the prior art still has deficiencies and needs to be improved.

[0005] Summary of the Invention

[0006] The embodiments of the present invention provide an animal experimental model and method for recording calcium signals, so as to achieve the purpose of dynamically recording and analyzing changes in calcium signals in animal neurons before and after intravenous drug delivery.

[0007] According to one embodiment of the present invention, an animal experimental model for calcium signal recording is provided, comprising the following steps:

[0008] introducing a preset fluorescent marker into neurons of the target body from a specific area of ​​the selected target body;

[0009] Implanting the autofocus module into a specific area;

[0010] Setting up a microscope camera mount in a specific area;

[0011] A microscope camera is arranged on a microscope camera fixing base for observing a specific area of ​​the target object;

[0012] One end of the drug delivery device is connected to the target's neck vein, and the other end is connected to the computer module. The computer module controls the drug delivery device to deliver the drug to the target and obtains data observed by the microscope camera.

[0013] In one embodiment, the self-focusing module is a glass self-focusing prism.

[0014] In one embodiment, the self-focusing module is fixed by light-curing resin.

[0015] In one embodiment, the model further comprises:

[0016] The camera data acquisition module is connected to the microscope camera at one end and the computer module at the other end. It is used to record data of a specific area and transmit it to the computer module.

[0017] In one embodiment, the model further comprises:

[0018] The nose touch panel is connected to the computer module, and is used to collect the nose touch signal of the target body and transmit a synchronization signal to the computer module.

[0019] In one embodiment, the model further comprises:

[0020] The nose touch data acquisition module is connected to the nose touch panel at one end and to the computer module at the other end. The nose touch panel sends a synchronization signal to the computer module through the data acquisition card.

[0021] An animal experimental method for calcium signal recording, comprising application of any of the above-mentioned animal experimental models for calcium signal recording, the method comprising:

[0022] Based on the drug administration signal, the computer module controls the drug administration device to administer the drug to the target body;

[0023] Use a microscope camera to observe a specific area of ​​the target object.

[0024] In one embodiment, after the computer module controls the drug delivery device to deliver the drug to the target based on the drug delivery signal, the method further includes:

[0025] The camera data acquisition module collects data of a specific area and transmits it to the computer module.

[0026] In one embodiment, after observing a specific area of ​​the target object using a microscope camera, the method further includes:

[0027] The nose touch module collects the target's nose touch signal in real time;

[0028] The data acquisition module sends a synchronization signal to the computer module based on the nose touch signal;

[0029] Based on the synchronization signal, the computer generates a synchronization file, which includes video data acquired by the microscope camera.

[0030] In one embodiment, after collecting the nose touch signal of the target body in real time through the nose touch module, the method further includes:

[0031] The nose contact data acquisition module drives the drug delivery device to deliver the drug to the target body based on the nose contact signal.

[0032] The animal experimental model and method for calcium signal recording in the embodiments of the present invention introduces a fluorescent marker into neurons in a selected target region, then mounts a microscopic camera in that region to observe brain activity. This invention establishes an animal model for intravenous drug administration and microscopic calcium signal recording. By observing changes in animal calcium signals at the moment of intravenous drug infusion, it facilitates subsequent investigations into drug effects and therapeutic mechanisms. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] FIG1 is a block diagram of an animal experimental model for calcium signal recording according to the present invention;

[0034] FIG2 is a schematic diagram showing the principle of the animal experimental model for calcium signal recording according to the present invention;

[0035] FIG3 is a schematic diagram of a target body of the present invention;

[0036] FIG4 is a schematic diagram of intravenous injection of the present invention;

[0037] FIG5 is a flow chart of the animal experimental method for calcium signal recording according to the present invention.

[0038] Figure numerals: 1-target body, 2-camera data acquisition module, 3-computer module, 4-nasal touch panel, 5-drug delivery device, 6-nasal touch data acquisition module, 7-drug delivery catheter, 8-microscope camera fixing seat, 9-self-focusing module. DETAILED DESCRIPTION

[0039] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts should fall within the scope of protection 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-mentioned drawings are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that the numbers used in this way can be interchanged where appropriate, 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 "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, system, product or device that includes a series of steps or units is not necessarily limited to those steps or units clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products or devices.

[0041] Example 1

[0042] According to one embodiment of the present invention, an animal experimental model for calcium signal recording is provided. Referring to FIG. 1 to FIG. 4 , the model includes:

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

[0044] Implanting the self-focusing module 9 into a specific area;

[0045] A microscope camera fixing seat 8 is provided in a specific area;

[0046] A microscope camera is provided on the microscope camera fixing base 8 for observing a specific area of ​​the target object 1;

[0047] One end of the drug delivery device 5 is connected to the neck vein of the target 1 , and the other end is connected to the computer module 3 . The computer module 3 controls the drug delivery device 5 to deliver the drug to the target 1 and obtains data observed by the microscope camera.

[0048] The present invention introduces a fluorescent marker into neurons of a selected target 1 from a specific region of the target 1, then positions a microscopic camera mount 8 at the specific region of the target 1, using the microscopic camera to observe brain activity in the target 1. The present invention establishes an animal model for intravenous drug administration and microscopic calcium signal recording. By observing changes in animal calcium signals at the moment of intravenous drug infusion, subsequent research into drug effects and therapeutic mechanisms is facilitated.

[0049] Specifically, the target 1 can be a mouse for experiment. The selected fluorescent marker or gene is introduced into the mouse neurons through gene transduction. This can be achieved through viral vectors (such as AAV) + transgenic mice to ensure sufficient expression in the target neurons. Operation steps: Anesthetize the mouse with anesthetic drugs, remove the hair on the head, cut the scalp, level the mouse head, drill the skull surface of the specific brain area with a skull drill, and inject a specific fluorescently labeled virus into the specific brain area of ​​the mouse brain. Among them, the fluorescent marker can be selected as calcium ions.

[0050] In one embodiment, referring to Figures 1 and 3 , the autofocusing module 9 is a glass autofocusing prism. Specifically, a glass autofocusing prism (Lens) for microscopic imaging is implanted into a specific brain region (a selected specific area) of a mouse along the virus injection channel. The Lens is secured to the mouse's head using a light-curing resin. The lens's focal plane is adjusted using a microscope camera. A microscope camera mount 8 is installed on the mouse's head. During the experiment, the microscope camera is connected to the mount 8.

[0051] The mouse was anesthetized with anesthetic drugs, and the hair on the right side of the mouse's neck skin was removed with depilatory cream. After disinfection, the skin was incised to expose the external jugular vein. Referring to Figure 4, a silicone tube was implanted into the blood vessel. The other end of the silicone tube was connected to the drug delivery device 5 through the back incision. The preliminary model animal was constructed after the mouse woke up.

[0052] In one embodiment, referring to FIG1 and FIG2 , the model further includes:

[0053] The video data acquisition module 2 is connected to a microscope camera at one end and to the computer module 3 at the other end, and is used to record data of a specific area 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, which processes and analyzes the nerve cells in the specific area.

[0055] In one embodiment, referring to FIG1 and FIG2 , the model further includes:

[0056] The nose touch panel 4 is connected to the computer module 3 and is used to collect the nose touch signal of the target body 1 and transmit a synchronization signal to the computer module 3.

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

[0058] In one embodiment, referring to FIG1 and FIG2 , the model further includes:

[0059] The nose touch data acquisition module 6 has one end connected to the nose touch panel 4 and the other end connected to the computer module 3. The nose touch panel 4 sends a synchronization signal to the computer module 3 through the data acquisition card.

[0060] Example 2

[0061] According to another embodiment of the present invention, an animal experimental method for recording calcium signals is provided, as shown in FIG1 , FIG2 and FIG5 , comprising:

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

[0063] S200: Observe a specific area of ​​the target object 1 using a microscope camera.

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

[0065] In one embodiment, referring to FIG1 and FIG2 , after the computer module 3 controls the drug delivery device 5 to deliver the drug to the target 1 based on the drug delivery signal, the following steps are further included:

[0066] The specific area data is collected by the camera data collection module 2 and transmitted to the computer module 3.

[0067] This application uses a microscope camera to observe the brain activity of the target body 1, realizes the construction of an animal model for intravenous drug administration and microscopic calcium signal recording, observes the changes in the animal's calcium signal at the moment of intravenous drug infusion through the microscope camera, and sends the observed signal to the computer module 3 for analysis, which facilitates subsequent exploration of drug effects and treatment mechanisms.

[0068] In one embodiment, referring to FIG1 and FIG2 , after observing a specific area of ​​the target object 1 using a microscope camera, the method further includes:

[0069] The nose touch module collects the nose touch signal of the target 1 in real time;

[0070] The data acquisition module sends a synchronization signal to the computer module 3 based on the nose touch signal;

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

[0072] Specifically, during the experiment, the mouse can touch the nose touch panel 4. Based on the mouse's touch, the nose touch panel 4 will transmit the nose touch signal to the nose touch data acquisition module 6, and the nose touch data acquisition module 6 will transmit it to the computer module 3 based on the received nose touch surface signal. At the same time, the receiving computer module 3 drives the drug administration signal to be sent 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 FIG1 and FIG2 , after the nose touch module collects the nose touch signal of the target 1 in real time, the method further includes:

[0074] Based on the nose contact signal, the data acquisition module drives the drug delivery device 5 to deliver the drug to the target 1 .

[0075] The computer module 3 will obtain the nose touch data from the nose touch data acquisition module 6. The nose touch data includes the time point and the number of times. Then the drug delivery device 5 will be controlled to deliver the drug according to the nose touch data, so that the drug delivery device 5 delivers the drug to the mouse based on the mouse's touch, and then the mouse will be repeatedly observed through the microscope camera.

[0076] With reference to Figures 1 and 2, the model principles and application methods of this application are as follows:

[0077] Model mouse (i.e., target body 1): A microscope camera fixing seat 8 is provided on the mouse head. During the experiment, the microscope camera is set on the microscope camera fixing seat 8 to connect the mouse head to the microscope camera, and then connected to the drug delivery device 5 through the white drug delivery tube on the back of the mouse.

[0078] The camera data acquisition module 2 is connected to the micro camera and the computer module 3, and is 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 microscope 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 the drug to the mouse through the drug delivery catheter on the mouse's back based on the 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. The synchronization signal can enable the computer module 3 to generate a synchronization file from the video signal of the microscope camera.

[0081] Drug delivery device 5: uses a micro-pump to deliver drugs to the model mouse through the drug delivery catheter after receiving the drug delivery signal from the nose contact data acquisition module 6.

[0082] Nose touch data acquisition module 6: used to connect the drug delivery device 5 and the nose touch panel 4 with the computer module 3. When receiving the nose touch signal from the nose touch panel 4, the nose touch signal is transmitted to the computer module 3. At the same time, the driving drug delivery signal fed back by the computer module 3 is received and sent to the drug delivery device 5. After receiving the feedback drug delivery signal, the drug delivery device 5 delivers the drug to the mouse.

[0083] This application selects a suitable calcium ion indicator and then transports the calcium ion indicator to a specific area of ​​the mouse brain. The calcium ion indicator is introduced into the target brain region through viral infection, allowing the activity of the infected neurons (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) to be observed under a fluorescence microscope (microscope camera). The microscope camera captures and records the changes in calcium signals, records the results, and analyzes the recorded data.

[0084] The purpose of the present invention is to construct an experimental animal model of intravenous catheterization combined with microscopic imaging, which has achieved the purpose of dynamically recording the changes in animal neuronal calcium signals before and after intravenous drug delivery. This animal model can be used to carry out a large number of brain science-related drug treatment mechanisms, and can be used for subsequent correlation research on anesthetic drugs, neurological diseases (such as Alzheimer's disease, epilepsy, etc.) and drug treatment.

[0085] The above are only preferred embodiments of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection 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 from a specific region of the selected target; 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; Setting one end of a drug delivery device to be connected to the jugular vein blood vessel of the target and the other end to be connected to a computer module, controlling the drug delivery device to administer drugs to the target 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 with a 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, wherein The model further includes: A nose touch panel, connected to the computer module, for collecting the nose touch signal of the target 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 experiment 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; Using a microscope camera to observe the specific region of the target.

8. The animal experiment 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 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 experiment 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, it further includes: Real-time collecting the nose touch signal of the target 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 acquired by the microscope camera.

10. The animal experiment method for calcium signal recording according to claim 8, wherein After real-time collecting the nose touch signal of the target 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 based on the nose touch signal.

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