Animal energy metabolism detection and real-time feedback regulation device and application thereof

Abstract

The invention relates to an animal energy metabolism detection and real-time feedback regulation device and application thereof. The device comprises a metabolism cabin, a monitoring system, an analysis feedback system and a regulation and control system. The metabolism cabin and the monitoring system are used for culturing and monitoring the energy metabolism condition of animals in real time; the analysis feedback system is responsible for receiving, storing and analyzing data transmitted by the monitoring system and providing control signals, and the regulation and control system is a wireless optical genetic regulation and control system and can regulate and control electrical activity of specific neurons of animals in real time so as to regulate energy metabolism, so that the time resolution and the cell specificity of animal energy metabolism research are improved.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology and relates to an animal energy metabolism detection and real-time feedback regulation device and its application. Background Technology

[0002] Respiratory entropy is an important indicator commonly used to reflect the real-time metabolic status of animals. It is defined as the ratio of the volume of oxygen consumed to the volume of carbon dioxide produced by the animal (VCO2 / VO2). Generally speaking, a respiratory entropy of 1 indicates that the metabolic substrate is mainly glucose, while a respiratory entropy of 0.7 indicates that the metabolic substrate is fat.

[0003] To determine the energy metabolism of laboratory animals and the effects of drugs on energy metabolism, many companies have launched respiratory entropy detection instruments. This article uses the CLAMS system (Comprehensive Lab Animal Monitoring System) from Columbus, USA, as an example to introduce the principle of a common small animal metabolic monitoring system. The technical solution is as follows: Mice are fasted overnight before the experiment. The weight is measured the next day. Each measurement cage is provided with bedding, drinking water, and rodent food, and the mice are placed in the metabolic cages for individual housing. Before calibrating the air measurement system, the silica gel in the desiccant is replaced to absorb moisture from the air entering the probe. The air intake speed is adjusted, and the valves for pure nitrogen and the oxygen / carbon dioxide / nitrogen mixture are opened to calibrate the oxygen and carbon dioxide measurement probes in the metabolic cages. The pure nitrogen valve is opened to calibrate the zero point of the oxygen measurement probe, and the offset knob is adjusted to bring the oxygen reading to approximately zero. The pure nitrogen valve is closed, and the valve for the 21.50% oxygen and 0.50% carbon dioxide mixture is opened to calibrate the upper limit of the oxygen probe measurement. The span offset is adjusted to bring the oxygen reading to approximately 21.50%. The calibration of the infrared carbon dioxide probe is completed by computer, requiring no manual adjustment. Once both probes are calibrated, 24-hour metabolic monitoring can begin. Oxygen consumption and carbon dioxide emissions are calculated based on the difference between the concentrations in the air and in the metabolic measurement cage, as well as the airflow velocity. Respiratory entropy (RER) is calculated based on the ratio of carbon dioxide excretion (VCO2) to oxygen consumption (VO2), typically between 0.7 and 1 (i.e., VCO2 / VO2 when fat and glucose are used as metabolic substrates). Total energy expenditure is calculated as (3.815 + 1.232 × VCO2 / VO2) × VO2 and averaged based on mouse body weight, thus detecting the effect of changes in specific experimental conditions on animal energy metabolism.

[0004] However, existing metabolic cages are mainly used for monitoring and cannot achieve real-time regulation of the metabolism of laboratory animals. Since metabolic cage detection requires a relatively closed box to record gas exchange, when it is necessary to regulate the energy metabolism of laboratory animals, existing methods include pre-experiment administration (or stimulation) or interruption of experimental administration. These methods not only add interference factors to the experiment, but also lack time resolution and cannot achieve automated operation.

[0005] In conclusion, developing a system that can regulate animal energy metabolism in real time is of great significance. Summary of the Invention

[0006] In view of the shortcomings of existing technologies and practical needs, the present invention provides an animal energy metabolism detection and real-time feedback regulation device and its application.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] In a first aspect, the present invention provides an animal energy metabolism detection and real-time feedback regulation device, the device comprising a metabolic chamber, a monitoring system, an analysis and feedback system, and a regulation system;

[0009] The metabolic chamber includes a metabolic cage and an antenna for culturing experimental animals;

[0010] The monitoring system includes an oxygen concentration detection module, a carbon dioxide concentration detection module, and an air flow rate detection module, used to detect the oxygen and carbon dioxide concentrations and flow rate of the air in the metabolic cage.

[0011] The analysis and feedback system includes a calculation module and a feedback module. The calculation module is used to collect oxygen and carbon dioxide concentrations and air circulation rate obtained by the monitoring system, and calculate the volume of oxygen consumed and carbon dioxide produced by the animal per unit time, respiratory entropy and energy consumption. The feedback module outputs control signals to the control system through the antenna.

[0012] The control system includes a signal receiving module and a light source emitting module. The signal receiving module is used to receive the control signal output by the feedback module and control the light source emitting module.

[0013] This invention designs a device for real-time detection and regulation of animal energy metabolism. It uses a metabolic chamber and a monitoring system to cultivate and monitor the energy metabolism of animals in real time, such as activity level, water intake and food intake, as well as indicators such as respiratory entropy and energy consumption. The analysis and feedback system is responsible for receiving, storing and analyzing the data transmitted by the monitoring system and providing control signals. The regulation system is a wireless optogenetic regulation system that can regulate the electrical activity of specific neurons in animals in real time to regulate energy metabolism.

[0014] Optionally, the metabolic cage is provided with at least two ventilation valves, at least one of which is configured to communicate with the ambient atmosphere when open, and at least one of which is configured to communicate with the monitoring system when open.

[0015] Optionally, the antenna is configured to be wound around the outer or inner wall of the metabolic cage.

[0016] Optionally, the oxygen concentration detection module includes a zirconia oxygen analyzer.

[0017] Optionally, the carbon dioxide concentration detection module includes an NDIR (non-dispersive infrared) self-diffusion carbon dioxide detector.

[0018] Optionally, the airflow rate detection module includes an air flow meter.

[0019] Optionally, the feedback module is used to compare the respiratory entropy and / or energy consumption calculated by the calculation module with a preset threshold, and output a control signal to the control system through the antenna according to the comparison result.

[0020] Optionally, the feedback module outputs radio frequency current through an antenna.

[0021] Optionally, the signal receiving module includes a coil, and the light source emitting module includes an LED light source;

[0022] Optionally, the coil can respond to the radio frequency current output by the antenna, thereby controlling the LED light source.

[0023] Secondly, the present invention provides the application of the animal energy metabolism detection and real-time feedback regulation device described in the first aspect in regulating and detecting animal energy metabolism.

[0024] Thirdly, the present invention provides a method for detecting and regulating animal energy metabolism, the method comprising:

[0025] The detection and regulation are performed using the animal energy metabolism detection and real-time feedback regulation device described in the first aspect.

[0026] Optionally, the method specifically includes:

[0027] A photosensitive gene is expressed in neurons in the brain region of experimental animals, which are then cultured in the metabolic cage. The oxygen and carbon dioxide concentrations and flow rates of the air in the metabolic cage are detected using the monitoring system. The respiratory entropy and / or energy consumption of the experimental animals are calculated using the analysis feedback system, compared with a preset threshold, and a control signal is output to the control system to turn the light source on or off.

[0028] Optionally, the experimental animals include any one of mice, rats, or rabbits.

[0029] Optionally, the light-sensitive gene includes the light-sensitive channel protein ChR2 gene or the rhodopsin NpHR gene from halophilic bacteria.

[0030] Optionally, expressing the light-sensing gene includes infecting neurons in the brain region of experimental animals with a viral vector containing the light-sensing gene.

[0031] Compared with the prior art, the present invention has at least the following beneficial effects:

[0032] This invention designs a device and method for real-time detection and regulation of animal energy metabolism. It utilizes a metabolic chamber and monitoring system to culture and monitor the energy metabolism of animals in real time, including indicators such as activity level, water intake and food consumption, as well as respiratory entropy and energy consumption. The analysis and feedback system is responsible for receiving, storing and analyzing the data transmitted by the monitoring system and providing control signals. The regulation system is a wireless optogenetic regulation system, which can regulate the electrical activity of specific neurons in animals in real time to regulate energy metabolism, thereby improving the temporal resolution and cell specificity of animal energy metabolism research. Attached Figure Description

[0033] Figure 1 This is a schematic diagram of the principle of an animal energy metabolism detection and real-time feedback regulation device.

[0034] Figure 2 This is a schematic diagram of the results of an animal energy metabolism detection and real-time feedback control device. 1 is the metabolic cage, 2 is the valve, 3 is the detection system, 4 is the analysis and feedback system, 5 is the control system, and 6 is the antenna.

[0035] Figure 3 This is an example diagram for detecting and regulating energy metabolism in mice in real time.

[0036] Figure 4 The figure shows the results of energy metabolism detection and real-time feedback regulation in mice. Detailed Implementation

[0037] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. However, the following examples are merely simplified examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention is determined by the claims.

[0038] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased from legitimate channels.

[0039] This invention provides a device for real-time regulation of animal energy metabolism, as shown in the schematic diagram below. Figure 1As shown, the metabolic chamber and monitoring system monitor the animal's energy metabolism in real time, such as activity level, water intake and food intake, as well as indicators such as respiratory entropy and energy consumption; the analysis and feedback system is responsible for receiving, storing and analyzing the data transmitted by the monitoring system and providing control signals; the regulation system is a wireless optogenetic regulation system, which can regulate the electrical activity of specific neurons in the animal in real time to regulate energy metabolism.

[0040] Based on the design and description of the device in this invention, those skilled in the art can understand and utilize various forms of corresponding hardware, software, firmware, dedicated processors, or combinations thereof in the art to obtain the device of this application.

[0041] In a specific embodiment of the present invention, such as Figure 2 As shown, a 40 cm × 18 cm × 20 cm acrylic testing chamber can be used as a metabolic cage. The chamber wall is 8 mm thick, and the outer wall has a recessed area with a snap-fit ​​antenna to fix the antenna to the outer wall. Specifically, the antenna can use ordinary copper wire (as long as the magnetic field strength is sufficient to light the light source). The chamber is sealable and has two ventilation valves, one connected to the ambient atmosphere and the other to the detection system. The connection points are equipped with ammonia removal and dust removal devices. A miniature air pump delivers gas from the chamber to the detection probe. A zirconia oxygen analyzer and an NDIR non-dispersive infrared self-diffusion carbon dioxide detector can be used to measure the oxygen and carbon dioxide concentrations in the ambient air and the air inside the chamber, respectively. An air flow meter measures the airflow rate. These data are transmitted to an analysis feedback system (which can use a computer capable of calculation and outputting control signals) to calculate the volume of oxygen consumed and carbon dioxide produced by the animal per unit time, i.e., the measurable respiratory entropy. By comparing with a set threshold, a control signal is transmitted to the control system. This control signal can be implemented based on radio frequency current.

[0042] The control system can use components capable of photostimulating experimental animals. The photosensitizing gene is expressed on neurons related to metabolic regulation in the hypothalamus of the experimental animals (this can be achieved through infection using a viral vector containing the photosensitizing gene). Specifically, the control system can use a commercially available ultra-miniature fully implantable μLED light source coil implant (purchased from Hangzhou Inper Technology, model A1). The implant integrates a receiver, μLED, and indicator light. The implant coil is fixed to the surface of the skull, and the red light emitted by the indicator light can penetrate the skin to display the stimulation status. Figure 3The coil has a biocompatible coating and a waterproof design, allowing for long-term stable reception of electromagnetic field signals. When the analysis feedback system transmits a "start" signal to the control system, an alternating magnetic field is generated within the metabolic cage using the principle of radio frequency power. The coil generates enough power to light the μLED. By controlling the radio frequency current, the μLED light source can be controlled, regulating neuronal activity and thus influencing the animal's energy metabolism. The analysis feedback system allows for the setting of control thresholds, as well as the frequency and interval of μLED flashing. Once the respiratory entropy and other values ​​return to levels below the set thresholds, the control system transmits a "stop" signal, and the control system ceases operation.

[0043] In a specific embodiment of the present invention, a mouse is used as an example to verify the technical solution of the present invention.

[0044] Respiratory entropy is the ratio of the volume of oxygen consumed to the volume of carbon dioxide produced (VCO2 / VO2).

[0045] The total calories consumed can be calculated using (3.815 + 1.232 × VCO2 / VO2) × VO2 and averaged based on the mouse's body weight.

[0046] Example 1

[0047] This embodiment performs real-time reading and regulation of animal energy metabolism.

[0048] The specific experimental procedure includes:

[0049] 1) Mice were randomly divided into control and experimental groups. After anesthesia, their heads were fixed using a stereotaxic device. The photosensitive gene vector NpHR (purchased from Shenzhen Brinkes Biotechnology Co., Ltd.) was injected into the hypothalamus of the experimental group mice. The control group mice were given empty vectors without the photosensitive gene. After injection, a yellow LED light source was implanted in the same brain region. The micro-control system was fixed to the head of the mice using tissue glue (3M, USA, VETBOND). The photosensitive gene was fully expressed after 21 days.

[0050] 2) Before the experiment, wind the antenna counterclockwise around the outside of the metabolic cage and place a single animal inside the metabolic cage (e.g., Figure 3 As shown, a biocage cover with a valve (connected to the detection system) is fixed above the metabolic cage. The valve allows gas to enter and exit, and the detection system detects the difference in oxygen and carbon dioxide concentrations in the gas. The analysis feedback system calculates metabolic indicators in real time.

[0051] 3) Set a threshold. If the metabolic index reaches the set level, control the regulation system to activate the light source at a frequency of 0.1 Hz. This can regulate the activity of hypothalamic neurons in the experimental group mice (no effect on the control group). The activity of these neurons can directly regulate the metabolic process and be read out by the detection system.

[0052] The results are as follows Figure 4As shown, the hypothalamus of the control group mice did not express the photosensitive gene, so the respiratory entropy and energy metabolism level of the animals did not change significantly after regulation using this system. However, the hypothalamus of the experimental group mice expressed the photosensitive gene. After the respiratory entropy detection reached the set threshold, regulation was performed, and a significant decrease in respiratory entropy and energy metabolism level were observed.

[0053] In summary, this invention designs an animal energy metabolism detection and real-time feedback regulation device and method. It utilizes a metabolic chamber and monitoring system to culture and monitor the energy metabolism of animals in real time. The analysis and feedback system receives, stores, and analyzes the data transmitted by the monitoring system and provides control signals. It can autonomously set regulation parameters, such as stimulation parameters (setting stimulation frequency and time), trigger parameters (setting the threshold for starting regulation), and stimulation delay (setting the delay time before stimulation begins). The regulation system is a wireless optogenetic regulation system, utilizing radio frequency power principles. It eliminates the need for heavy equipment such as implanted batteries. Combined with wireless optogenetic technology, the implant structure is extremely small and lightweight. It achieves regulation of animal energy metabolism without requiring fiber optic connections or affecting the animal's free movement, and without interrupting monitoring or compromising the overall airtightness of the testing chamber. Furthermore, the flexible and waterproof design of the implant makes it suitable for the central and peripheral nervous systems and organs, and can be applied to research in neural circuit function, diseases (anxiety and depression, movement disorders), tumors, cell biology, and other fields.

[0054] The applicant declares that the above description is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention fall within the protection and disclosure scope of the present invention.

Claims

1. An animal energy metabolism detection and real-time feedback regulation device, characterized in that, The device comprises a metabolic chamber, a monitoring system, an analysis feedback system and a regulation system; The metabolic chamber comprises a metabolic cage and an antenna, and is used for culturing test animals; The monitoring system comprises an oxygen concentration detection module, a carbon dioxide concentration detection module and an air flow rate detection module, and is used for detecting the oxygen and carbon dioxide concentrations and the flow rate of air in the metabolic cage; The analysis feedback system comprises a calculation module and a feedback module, the calculation module is used for collecting the oxygen and carbon dioxide concentrations and the air flow rate obtained by the monitoring system, calculating the oxygen consumption and carbon dioxide production volume of the animals per unit time, the respiratory entropy and the energy consumption, and the feedback module outputs a control signal to the regulation system through the antenna; The regulation system comprises a signal receiving module and a light source emitting module, and the signal receiving module is used for receiving the control signal output by the feedback module and controlling the light source emitting module.

2. The animal energy metabolism detection and real-time feedback regulation device according to claim 1, characterized in that, The metabolic cage is provided with at least two ventilation valves, at least one of which is arranged to be in communication with the ambient atmosphere when opened, and at least one of which is arranged to be in communication with the monitoring system when opened; Optionally, the antenna is arranged to be wound around the outer wall or the inner wall of the metabolic cage.

3. The animal energy metabolism detection and real-time feedback regulation device according to claim 1 or 2, characterized in that, The oxygen concentration detection module comprises a zirconia oxygen analyzer; Optionally, the carbon dioxide concentration detection module comprises an NDIR non-dispersive infrared self-diffusion carbon dioxide detector; Optionally, the air flow rate detection module comprises an air flow meter.

4. The animal energy metabolism detection and real-time feedback regulation device according to any one of claims 1-3, characterized in that, The feedback module is used for comparing the respiratory entropy and / or energy consumption calculated by the calculation module with a preset threshold, and outputting a control signal to the regulation system through the antenna according to the comparison result.

5. The animal energy metabolism detection and real-time feedback regulation device according to any one of claims 1-4, characterized in that, The feedback module outputs a radio frequency current through the antenna.

6. The animal energy metabolism detection and real-time feedback regulation device according to any one of claims 1-5, characterized in that, The signal receiving module comprises a coil, and the light source emitting module comprises an LED light source; Optionally, the coil can respond to the radio frequency current output by the antenna, thereby controlling the LED light source. Optionally, the regulation system comprises a coil implant body of an implanted μLED light source.

7. The animal energy metabolism detection and real-time feedback regulation device according to any one of claims 1-5 is applied to regulating and detecting animal energy metabolism.

8. A method of detecting and regulating energy metabolism in an animal, characterized in that, The method comprises: detecting and regulating by using the animal energy metabolism detection and real-time feedback regulation device according to any one of claims 1-5.

9. The method of detecting and modulating energy metabolism in an animal according to claim 8, wherein, The method specifically comprises: expressing a light-sensitive gene in neurons in the brain region of the experimental animal, placing the experimental animal in the metabolic cage for culturing, detecting the oxygen and carbon dioxide concentrations and the flow rate of air in the metabolic cage by using the monitoring system, calculating the respiratory entropy and / or energy consumption of the experimental animal by using the analysis feedback system, comparing the respiratory entropy and / or energy consumption with a preset threshold, and outputting a control signal for turning on or turning off the light source emitting to the regulation system.

10. The method of detecting and regulating energy metabolism in an animal according to claim 8 or 9, wherein, The experimental animal comprises any one of a mouse, a rat or a rabbit; Optionally, the light-sensitive gene comprises a channelrhodopsin ChR2 gene or a halorhodopsin NpHR gene; Optionally, the expression of the light-sensitive gene comprises infecting neurons in the brain region of the experimental animal by using a viral vector containing the light-sensitive gene.

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