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Survey of research methods for swimming in large mice
Survey of research methods for swimming in large mice
Source: http://
〠Abstract】 In this paper, the methods of swimming experiments in large mice (forced swimming, weight-bearing swimming, water maze, etc.), evaluation indicators (inactive time, weight-bearing swimming time, escape latency, etc.) and the changes of physiological and biochemical characteristics of the model and mice were carried out. In summary, for the study of antidepressant, anti-fatigue and puzzle drugs, this paper also briefly introduces the application of computer technology in large mouse swimming experiments.
〠Key words】 Rat, mouse, swimming
The large mouse swimming experiment is an anti-stress experimental method widely used in the study of anti-depression, anti-fatigue and nootropic drugs. This article provides a brief overview of this, providing a reference for animal experiments on related drugs.
Application of 1 large and mouse swimming experiment in the study of antidepressant drugs
In 1977, Porsolt RD first applied a forced swimming test to detect the effects of antidepressants. Later forced swimming experiments became an animal model for evaluating the antidepressant effects of drugs. The experimental method is a behavioral desperation experiment method. The basic principle is that when a rat or a mouse is put into a limited space to swim, it is desperate to swim and try to escape at the beginning, and soon becomes a floating state, only exposed. The nostrils keep breathing, and the limbs are occasionally swiped to keep the body from sinking. It is actually the hope that the animal gives up the escape and is desperate.
The classic forced swimming experiment was conducted in two days. On the first day, the rats or mice were forced to swim for 15 min in deep water at 25 ° C. After being taken out, they were dried in a greenhouse at °C and returned to the cage. On the second day, forced swimming was performed for 5 minutes under the same conditions, and the immobility time was recorded. Mirua H et al. changed the time of forced swimming to (10-30) mini' 1 . On the second day of the experiment, the immobility time map of the mice was observed for 4 min after 6 min of forced swimming. These are the repairs made to the classic experimental methods.
change. The immobility time of rats or mice is to judge the antidepressant effect
index of. The shorter the immobility time, the stronger the antidepressant effect.
Water temperature, water depth, animals, and seasons are four factors that affect the experiment. Kitada et al. pointed out that a water temperature below 20 °C would shorten the immobility time. Therefore, the temperature of the water is often controlled at 25-30 OC [s7]. The choice of water depth should be based on the fact that animals cannot escape. In general, the experimental experiment in rats is to improve the imbalance of amino acid transmitters in the brain of mice with depression, increase the ratio of excitatory/inhibitory amino acids, reduce the time of forced swimming, increase the excitability of mice, and resist forced swimming. Depression tendency [9] 0
Application of 2 large mouse swimming experiments in anti-fatigue drugs research
The weight-bearing swimming test is an animal model for evaluating the anti-fatigue effect of drugs. OMoriura et al. used this experiment to evaluate the anti-fatigue effect of 50% ethanol extract of python (scientific name Agkistrodon blomhoffii blomhoffii Boie). [0] The commonly used experimental method is small. The mouse tail was loaded at 1/3 to 2/3 (5 to 10% of body weight) and then swallowed in a sink to observe and record the mouse from human water to sink into the water 10 . Time that cannot rise to the surface (in a state of exhaustion). This weight-bearing swimming time is regarded as an indicator for quantitatively evaluating the degree of fatigue. The longer the weight-bearing swimming time, the better the anti-fatigue effect of the drug. In the experiment, the water temperature (25-29 soil 1) OC, the water depth is about 25cm, and the weight is the lead wire or the plasticine ball. No weight-bearing swimming can also fatigue the mice, but it takes a long time to observe that it is in a state of exhaustion. A rat swimming fatigue model was established by placing the model rats in a water bath with a temperature of (43 ± 0.5) 'C and a water depth of 35 cm, allowing them to swim naturally, with natural sedimentation in each rat. The time was the fatigue time of each rat. When the whole group of 50% rats showed natural sedimentation, the whole group of animals stopped swimming and the modeling was continued for 14 days. After successful model, the rats showed: the stool frequency increased slightly, the lean, the weight was slightly
Decline, free activity is reduced, the coat is slightly fluffy, and the food intake is reduced. In recent years, forced swimming experiments have also been used for endurance testing.
If the water temperature is adjusted to (5 dexterous soil 1) °C or (39-45 ± 1) `C during the experiment, the mice can swim at high or low temperatures, and the effects of drugs on fatigue and stress caused by swimming can be examined. Use strong cooling water swimming as a stress source, mold at different times of the day, change the duration of daily modeling, reduce the body's tolerance to stress, and try to create a temperature stimulus (cold), physical exertion (swim) and spirit An animal model of stress caused by a variety of stress factors (stimuli stimulated by strong cooling water swimming). It not only simulates the pathogenesis of fatigue in Chinese medicine, but also meets the theory of chronic stress caused by Western medicine. The model animals have decreased physical strength (short exhausted swimming time), depression tendency (reduced spontaneous activity), and neuroendocrine dysfunction, which are similar to the symptoms and laboratory tests of fatigue patients. It can be seen that the model is an animal model that can be further improved and improved to study chronic fatigue. It also shows that the process of stress-induced chronic fatigue is closely related to the disorder of the body's neuro-endocrine-immune system. It was found that the concentration of monoamine transmitters in the serum of rats was generally increased, and the content of dopamine was significantly increased. Xiaoyi Yishen Oral Liquid (an anti-fatigue drug) can reduce the content of 5-tryptamine (5-HT), dopamine (DA) and its metabolite high vanillic acid (HVA), and make 5-HT metabolite 5 - The content of 5-HAA was increased, suggesting that its anti-stress effect may be related to the inhibition of the synthesis and metabolism of peripheral DA and the promotion of 5-HT decomposition. Strong cooling water swimming can also significantly reduce the ascorbate content of the rat adrenal gland (a classic indicator of adrenal function), indicating that adrenal function is active and corticosterone synthesis is increased. Xiaowei Yishen Oral Liquid can effectively reverse this change, suggesting that it can inhibit the hyperactivity of adrenal cortex function under stress, reduce the synthesis of adrenocortical hormone, and reduce the adverse effects caused by stress. Lactic acid (LAC), lactate Hydrogenase (LDH), serum urea nitrogen (BUN), glucose (Glc) and total protein (TP) are blood biochemical indicators associated with swimming fatigue. The accumulation of LAC is an important cause of exercise fatigue. As a product of anaerobic glycolysis, LAC will cause LAC accumulation in the body and affect the stability and normal metabolism of the body environment. LDH plays a catalytic role in the metabolic process of LAC, and the increase of its activity during the recovery period after exercise is beneficial to the elimination of LAC. BUN is a product of material metabolism during exercise. It increases with the increase of labor and exercise load. The worse the body adapts to load, the more obvious the increase of BUN. Exist in muscle, red blood cells and other tissues and is one of the sources of energy for exercise. In addition, the protein content can reflect the nutritional status of the body. The drug can achieve anti-fatigue effects by reducing LAC levels and BUN increments after exercise, increasing LDH activity and Glc and TP levels. Swimming experiments are frequently used in the study of anti-fatigue effects of traditional Chinese medicine. Liu Hong et al. used swimming mice as research objects and found that bamboo ginseng can increase LDH activity, reserve of muscle glycogen and liver glycogen, reduce LAC level and BUN increment after swimming, and have anti-fatigue effect. Bin Xiaonong et al. conducted pharmacological studies on Gynostemma pentaphyllum, and found that it can significantly prolong the time of swimming to exhaustion; reduce the content of malondialdehyde (MDA) in heart and kidney tissues of exhausted swimming mice; Superoxide dismutase (SOD) activity in the tissue increases the SOD/MDA ratio relatively. This suggests that Gynostemma pentaphyllum has the effect of improving exercise resistance against fatigue, heart and kidney free radical damage. Yang Ke et al pointed out that the ginseng diol saponin in ginseng can correct the significant decrease of serum ketone in rats caused by long-term increasing load swimming, and it is one of the mechanisms of anti-fatigue effect.
3 large, mouse swimming experiments in the study of educational drugs swimming experiments are often used in the study of learning and memory, including Morris, water maze, T-water maze, square water maze and Y-type water maze.
Since the invention of the Morris water mace (MWM) in 1981, MWM has become one of the main tools for scholars engaged in learning and memory research to detect spatial learning and memory in rodents. The basis of the learning and memory experiment method is conditioned reflex. The animals are measured or their response time to measure the storage capacity, retention time and the conditions on which they depend, after learning or performing a certain task. The Morris water maze consists of a circular pool and an automatic video recording system. The four equidistant points N, E, S, W on the pool wall are the starting points of the test. The sub-tank is 4 quadrants NE, SE, SW, NW. Optional one quadrant is placed in the center of the platform. Large and small mice swim here to climb the platform to get a rest. After many trainings, the memory can be obtained, and the learning and memory ability can be evaluated through behavioral testing. Whether large or small mice can determine that the platform is completely determined by their spatial learning ability, because the pool is circular, animals can only be positioned by reference clues around the pool. In the experiment, the reference clues were required to be rich and remain unchanged.
Two tests were often performed during the behavioral testing process with the Morris water maze:
(1) Positioning navigation test: The rats were placed in the pool 1 day before the start of the test (excluding the platform). Swim freely for 2 min to familiarize yourself with the maze environment. The test lasted for 5 days, and was divided into two periods, one time in the afternoon and two in the afternoon. Each time period was trained four times, 8 times a day. At the beginning of the training, the platform was placed in the NW quadrant, and the rat was placed in the pool wall from any of the four starting points. The automatic video recording system recorded the time (evacuation latency) and swimming path of the rat to find the platform, 4 times. The rats were trained to release water from four different starting points. After the rats found the platform or could not find the platform within 120s, the experimenter took the platform (the incubation period was 120s) and rested on the platform for 30s. Go to the next experiment.
(2) Space exploration test: On the fifth day of the fifth day (immediately after the fourth time), remove the platform, and select one person to point the rat to the pool wall and record the swimming in the pool within 1 min.
The number of strokes, the number of piercings (the number of times across the original platform) and the distance traveled in each quadrant as a percentage of the total distance.
Long Dahong et al. used the above experiments to discuss the effect of nerve growth factor (NGF) on the learning and memory ability of Alzheimer's disease rats. It was found that NGF can shorten the escape latency and increase the percentage of the distance between the number of piercing rings and the original platform quadrant. This suggests that NGF can improve the learning and memory ability of Alzheimer's rats.
At present, there is a more advanced, scientific and real Morris automatic water maze in the world. The device mainly has a labyrinth, a camera recording system and a computer system. The learning memory score is calculated by multiplying the time spent by the animal in each zone by the specified weighting value; the latency score refers to the number of seconds the animal needs to enter the maze to find the platform. Morris automatic water maze has the following advantages: There is Video camera system, which replaces the long-term visual observation of the researcher in the labyrinth test, and is not subject to any human factors error, more objective and real; using the microcomputer system and design program, not only can The test scores, the memory retention scores, and the animal's swimming trajectory are automatically displayed on the computer screen.
The learning and memory behaviors of mice in the circular water maze were converted into moving images and their real-time motion trajectory routes were displayed. The information resources obtained by the computer were comprehensively analyzed. After optimization and combination, the distance, speed and platform were established. An evaluation indicator consisting of time and trajectory. In the experiment, a test box with a height of 30cm and a diameter of 65cm was used. The bottom layer was equipped with a heating device, the temperature controller automatically controlled the water temperature (25 °C to 32 °C optional), and the white camera background (water surface color) in the Morris water maze method. Adjusted to black to simplify the experimental operation and ensure the accurate identification of the target, the multimedia video card is used instead of the dedicated image card to ensure the real-time and accuracy of signal acquisition.