CN113951200A - Multichannel device for detecting color preference of zebra fish juvenile fish and use method and application thereof - Google Patents
Multichannel device for detecting color preference of zebra fish juvenile fish and use method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
- A01K63/003—Aquaria; Terraria
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01K29/00—Other apparatus for animal husbandry
- A01K29/005—Monitoring or measuring activity, e.g. detecting heat or mating
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
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- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
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Abstract
The invention relates to the technical field of zebra fish juvenile fish behavior testing, and discloses a multichannel device for detecting zebra fish juvenile fish color preference and a using method and application thereof. The multi-channel tool comprises at least one transparent mold cuvette and at least two monochrome modules; all of the monochrome modules have at least two different colors; the monochromatic module is provided with a groove matched with the end part of the small transparent mould pool, and the end part of the small transparent mould pool can be arranged in the groove of the monochromatic module; the grooves on all the single-color modules are the same in shape and size. The invention can reduce accidental and artificial interference, and improve the reliability and detection flux of the detection result; meanwhile, by detecting the total movement distance of the zebra fish juvenile fish in different color areas, the preference of the zebra fish juvenile fish to a certain color can be quantitatively analyzed, and the transverse comparison among different groups of zebra fish juvenile fish and the longitudinal comparison of the zebra fish juvenile fish before and after a certain specific treatment are realized.
Description
Technical Field
The invention relates to the technical field of zebra fish behavior testing, in particular to a multichannel device for detecting color preference of zebra fish juvenile fish and a using method and application thereof.
Background
Cognition is an intelligent processing process for understanding and acquiring knowledge by organisms, and relates to a series of random, psychological and social behaviors such as learning, memory, language, thinking, spirit, emotion and the like. Cognitive impairment is a disease in which brain function is degraded due to brain neurocytopathy, and the memory, understanding, language, learning, calculation and judgment abilities of patients are affected, and there are some changes in mood, behavior and feeling. The factors which are commonly used at present and cause cognitive impairment mainly comprise age, genetic factors, brain diseases, trauma, medicinal factors and the like. The treatment for cognitive impairment varies depending on the causative factors, and is mainly based on drug therapy such as cerebral circulation-improving agent, energy metabolism activator, Ca, in addition to surgical therapy2+Antagonists, glutamate receptor antagonists, antioxidants, glial cell line modulators, and nonsteroidal anti-inflammatory agents, as well as neurotransmitter supplements.
The research and development of the drug can not be separated from biological experiments, namely the research of pathogenic mechanisms and the screening of candidate drugs until the final evaluation of drug effect, pharmacology and toxicology. In vitro and animal experiments play an important role in clarifying the mechanism of cognitive impairment and in the research and development of drugs. The in vitro experiment has simple operation and short period, but the behavioral phenotype related to cognitive impairment can not be observed. Although the traditional mammalian experiment can evaluate the phenotype related to the cognitive disorder through behavioral research, the experiment period is long, the flux is low, the cost is high, and the pathogenesis of the cognitive disorder and the development process of related medicaments are seriously hindered. Therefore, it is of great importance to find an alternative in vivo evaluation model and method for mammals.
Zebrafish are a novel model organism. The biological structure and physiological function of the medicine are highly similar to those of mammals, the development period is short, the size is small, the dosage is small, and the medicine is transparent and easy to observe. In addition, zebra fish can be screened in a multi-hole plate rapidly and at high flux, so that the zebra fish becomes a model organism with huge potential in the process of drug research and development. A large number of researches show that the zebra fish can be used for evaluating behaviors of high-class organisms such as learning, memory, cognition and the like, and a certain evaluation standard is formed. For example, zebra fish can distinguish different colors, and researches show that zebra fish juvenile fish selectively favors blue among four colors of blue, yellow, red and green. After bisphenol a (bpa) treatment, zebrafish neurotransmitter expression is abnormal, and behavioral and color preferences change, exhibiting a cognitive dysfunction phenotype. The above results indicate that BPA causes a disorder of zebrafish juvenile neurotransmitter by means of chemical damage, resulting in zebrafish exhibiting cognitive impairment. Another study showed that the homologue bisphenol f (bpf) of BPA also has an effect on zebrafish behavior by disrupting the expression of the juvenile zebrafish neurotransmitter. Therefore, the preference of the zebra fish juvenile fish for different colors can be evaluated to evaluate the cognitive ability of the zebra fish. Furthermore, the zebra fish juvenile cognitive impairment model has the potential for screening of neuroprotective agents in cooperation with a zebra fish color preference experiment.
However, in the prior art, the traditional four-color maze experiment is usually adopted to study the color preference of zebra fish, which has certain defects: 1) taking a picture of an experimental end point to determine the position where the zebra fish stays, wherein the picture only represents the instantaneous result of the exposure end point and has contingency; 2) zebra fish is susceptible to noise interference caused by photographing, so that results are inaccurate; 3) the cross maze experiment flux is less, wastes time and energy.
Disclosure of Invention
In order to solve the technical problem, the invention provides a multichannel instrument for detecting the color preference of zebra fish juvenile fish and a using method and application thereof. The invention can reduce accidental and artificial interference, improve the reliability of the detection result and improve the detection flux.
The specific technical scheme of the invention is as follows:
in a first aspect, the present invention provides a multichannel device for detecting colour preferences of zebra fish larvae, comprising at least one cuvette and at least two monochrome modules; all of the monochrome modules have at least two different colors; the monochromatic module is provided with a groove matched with the end part of the small transparent mould pool, and the end part of the small transparent mould pool can be arranged in the groove of the monochromatic module; the grooves on all the single-color modules are the same in shape and size.
When the multichannel device is used, the two monochromatic modules with different colors are installed at the two ends of the small transparent mould pool, and the grooves have certain depth, so that color areas with certain length can be formed at the two ends of the small transparent mould pool, therefore, by means of a zebra fish behavior monitoring instrument (such as a zebra fish behavior instrument Viewpoint which is mainstream in the current market), the total movement distance of the zebra fish juvenile fish in the two color areas within a certain time can be counted, and the color preference of the zebra fish juvenile fish can be further judged.
Compared with the traditional four-color cross maze experiment, the multichannel device disclosed by the invention is used for judging the color preference of the zebra fish juvenile fish, so that data can be automatically acquired for a long time, the personnel interference is avoided, and the result reliability is higher; and, through setting up a plurality of transparent mould cistocks, usable zebra fish action monitoring instrument tests a plurality of transparent mould cistocks simultaneously, realizes multiunit parallel experiment, makes the flux improve greatly.
Preferably, the number of monochrome modules is at least four, including at least one blue module, at least one yellow module, at least one red module and at least one green module.
Preferably, the small transparent mold pool is rectangular; the external width of the small transparent mould pool is not more than the opening width of the groove; and half of the external length of the cuvette of the transparent mould is not less than the opening depth of the groove.
Preferably, the external width of the transparent die cuvette and the opening width of the groove are both 11-17 mm.
Preferably, the external length of the transparent die cuvette is 30-38 mm, and the opening depth of the groove is 8-12 mm.
Preferably, the height of the inner part of the small transparent mould pool and the height of the groove are both 8-12 mm.
Preferably, the wall thickness of the small transparent mold pool is 1.5-2.5 mm.
According to the requirements of national standards, the ratio of the weight of the zebra fish to the volume of water in the processes of feeding and experiment of the zebra fish is not more than 1g/L, and the weight of 5-6 dpf of the zebra fish juvenile fish used in the invention is about 0.3mg, so that each fish needs 0.3mL of water, and 4-6 fish needs 1.2-1.8 mL. If the size is less than the above listed parameters, the experimental process does not meet the national standard requirements. In addition, according to the previous experimental observation of the inventor, the space where the zebra fish juvenile fish is located is too narrow, and the spontaneous movement behavior of the zebra fish juvenile fish is seriously influenced. The length of the zebra fish juvenile fish with 5-6 dpf is 4-5 mm, and if the size of the small pool is smaller than the parameters, spontaneous movement of the zebra fish is influenced, so that the experimental result is inaccurate. On the other hand, if the size is larger than the above parameters, the number of cuvettes will decrease, resulting in a decrease in experimental throughput.
In a second aspect, the invention provides a method for detecting zebra fish juvenile color preference by using the multichannel device, comprising the following steps:
(1) putting the zebra fish juvenile fish into a small transparent mould pool, and adding water into the small transparent mould pool;
(2) installing a single-color module at each of two ends of each transparent mold cuvette respectively, so that the single-color modules at the two ends of each transparent mold cuvette have different colors, namely A color and B color;
(3) after the zebra fish juvenile fish adapts to the environment in the transparent mold small pool, data collection is carried out, and the total movement distance of the zebra fish juvenile fish in different color areas is counted;
(4) and judging the color preference of the zebra fish juvenile fish according to the total movement distance of the zebra fish juvenile fish in different color areas.
According to the invention, the color preference of the zebra fish juvenile fish is judged by detecting the total movement distance of the zebra fish juvenile fish in different color areas within a certain time, and by adopting the method, the accidental shooting of the traditional four-color maze experiment end point can be overcome, meanwhile, the interference of personnel can be avoided, and the result is more reliable.
In addition, some prior arts (for example, patent CN202010887716.3) determine the color preference by detecting the latency of zebra fish in different color areas, and the present invention adopts the total moving distance compared to the latency, which has the following advantages: 1 o latency in different color zones is usually determined by artificially analyzing the residence time of the adult zebra fish in a certain area in an experimental video, and the method is susceptible to subjective judgment, and the experimental result has large errors. The total movement distance adopted by the invention is automatically and accurately quantified by the zebra fish behavior analyzer, and the result is reliable. 2, the incubation time in different color areas generally uses adult zebrafish, and the experimental process is complicated, the individual difference is large, and the flux is low. The method is suitable for the juvenile zebra fish, can monitor the behaviors of a plurality of zebra fish simultaneously, and has the advantages of simple experimental process, small individual difference and high flux.
Preferably, in the step (1), 4-6 zebra fish juvenile fishes are placed in each transparent die cell; the zebra fish juvenile fish is 5-6 days after fertilization.
If the number of the fishes is too large, the national standard requirements cannot be met, and the interference on the spontaneous behaviors of the zebra fish juvenile fishes is caused; too few fish may result in unstable data.
Preferably, in the step (3), the adaptation time of the zebra fish juvenile fish to the environment in the transparent mold cell is 8-15 min.
Preferably, in the step (3), the data acquisition adopts a continuous acquisition mode, and the data acquisition time is 10-15 min.
Preferably, in the step (3), the data acquisition adopts an interval acquisition mode, and the specific process is as follows: stopping data acquisition after 5-8 min of data acquisition, exchanging the single-color modules at two ends of the small transparent mould pool under the condition of not moving the small transparent mould pool, and then continuing to acquire data; and carrying out data acquisition for 2-4 times in total.
The inventor pays attention to the fact that the zebra fish juvenile fish possibly stops in the preferred color area and does not move during data collection, and errors occur in the detection result of the color preference. In order to solve the technical problem, the positions of the single-color modules are periodically changed in the data acquisition process, and the zebra fish juvenile fishes are promoted to swim through the change of the colors of the two ends of the small pool of the transparent mold, so that errors caused by the fact that the zebra fish juvenile fishes are still in the preferred color areas are reduced.
Preferably, the specific process of step (4) is as follows: calculating the ratio of the moving distances of the zebra fish juvenile in the color A area to the moving distances of the zebra fish juvenile in the color B area according to the total moving distances of the zebra fish juvenile in the color A area and the zebra fish juvenile in the color B area, and judging the preference of the zebra fish juvenile to the color A; the ratio of the movement distance of the color A region is calculated according to the following formula:
on the basis of obtaining the total movement distance of each color area, calculating the movement distance ratio of a certain color (such as color A) area, and by utilizing the movement distance ratio, the preference of the zebra fish juvenile fish to a certain color can be quantitatively analyzed, so that the transverse comparison among different groups of zebra fish juvenile fish and the longitudinal comparison of the zebra fish juvenile fish before and after a certain specific treatment can be realized, and accordingly, the strength of the zebra fish juvenile fish in different growth periods to the preference of a certain color, the influence of a certain treatment mode (such as medication) to the cognitive ability of the zebra fish juvenile fish, or the strength of the influence of different treatment modes (such as different medicines) to the cognitive ability of the zebra fish juvenile fish can be researched.
In a third aspect, the invention provides an application of the multichannel device or the method for detecting the color preference of the zebra fish juvenile fish in screening of the cognitive disorder treatment drugs.
By using zebra fish as a model organism and evaluating the cognitive ability of the zebra fish juvenile fish, screening of cognitive disorder treatment drugs such as neuroprotective agents and neurotransmitter supplements can be performed.
Compared with the prior art, the invention has the following advantages:
(1) data can be automatically acquired for a long time, so that personnel interference is avoided, and the result is more reliable;
(2) the preference of the zebra fish juvenile fish to a certain color can be quantitatively analyzed, and transverse comparison among different groups of zebra fish juvenile fish and longitudinal comparison of the zebra fish juvenile fish before and after a certain specific treatment are realized;
(3) can carry out multiunit parallel experiment simultaneously, the flux is high, is applicable to the detection of big batch sample.
Drawings
FIG. 1 is a top view of a multichannel instrument of the invention;
FIG. 2 is a top view of the monochrome module of FIG. 1; .
FIG. 3 is a pictorial view of a multichannel instrument of the invention; the drawing a is a main body of the mold, the drawing B is a monochrome module of 4 colors (the upper left is a green module, the lower left is a red module, the upper right is a blue module, and the lower right is a yellow module), and the drawing C is a transparent mold cuvette with monochrome modules mounted at both ends.
FIG. 4 shows the ratio of the moving distance of each color area of 5dpf and 6dpf zebra fish juvenile fish in the blue-yellow, blue-red, blue-green color combination; wherein, the graph A is the ratio of the moving distance of each color area of 5dpf zebra fish juvenile fish in each color combination, and the graph B is the ratio of the moving distance of each color area of 6dpf zebra fish juvenile fish in each color combination.
Fig. 5 shows the ratio of the moving distance of 6dpf zebra fish larvae in the blue-yellow color combination after BPAF molding and edaravone positive treatment in application example 2.
FIG. 6 shows the ratio of the moving distance of 6dpf zebra fish larvae in blue-yellow color combination after BPAF modeling and edaravone positive treatment in application example 3.
The reference signs are: the device comprises a small transparent mould pool 1, a single-color module 2, a groove 2-1 and a mould main body 3.
Detailed Description
The present invention will be further described with reference to the following examples.
General examples
A multichannel device for detecting the colour preference of zebra fish larvae comprises at least one rectangular transparent mould cuvette 1 and at least four monochrome modules 2. All the monochrome modules 2 include at least one blue module, at least one yellow module, at least one red module, and at least one green module. The monochromatic module 2 is provided with a groove 2-1 matched with the end part of the transparent mould cuvette 1, and the end part of the transparent mould cuvette 1 can be arranged in the groove 2-1 of the monochromatic module 2; the grooves 2-1 on all the monochrome modules 2 are the same in shape and size. The external width and the external length of the transparent die cuvette 1 are respectively 11-17 mm and 30-38 mm, the internal height is 8-12 mm, and the wall thickness is 1.5-2.5 mm; the opening width, the opening depth and the height of the groove 2-1 are respectively 11-17 mm, 8-12 mm and 8-12 mm; the external width of the transparent mould cuvette 1 is not more than the opening width of the groove 2-1, and half of the external length of the transparent mould cuvette 1 is not less than the opening depth of the groove 2-1.
A method for detecting zebra fish juvenile color preference using the multi-channel tool, comprising the steps of:
(1) putting 4-6 zebra fish juvenile fishes into each transparent mould cuvette, and adding water into the transparent mould cuvette;
(2) installing a single-color module at each of two ends of each transparent mold cuvette respectively, so that the single-color modules at the two ends of each transparent mold cuvette have different colors, namely A color and B color;
(3) after the zebra fish juvenile fish adapts to the environment in the transparent mold small pool, data collection is carried out, and the total movement distance of the zebra fish juvenile fish in different color areas is counted;
optionally, the data acquisition adopts a continuous acquisition mode, and the data acquisition time is 10-15 min; or, the data acquisition adopts an interval acquisition mode, and the specific process is as follows: stopping data acquisition after 5-8 min of data acquisition, exchanging the single-color modules at two ends of the small transparent mould pool under the condition of not moving the small transparent mould pool, and then continuing to acquire data; data acquisition is carried out for 2-4 times in total;
(4) and judging the color preference of the zebra fish juvenile fish according to the total movement distance of the zebra fish juvenile fish in different color areas.
The multichannel device or the method for detecting the color preference of the zebra fish juvenile fish is applied to evaluation of cognitive ability of the zebra fish juvenile fish.
Example 1
As shown in fig. 1 and 2, a multichannel device for detecting color preference of zebra fish juvenile fish includes a mold body 3 and a monochrome module 2. 12 rectangular transparent mold small pools 1 are arranged on the upper surface of the mold main body 3. The monochrome module 2 includes 12 blue modules, 12 yellow modules, 12 red modules, and 12 green modules. The monochromatic module 2 is provided with a groove 2-1 matched with the end part of the transparent mould cuvette 1, and the end part of the transparent mould cuvette 1 can be arranged in the groove 2-1 of the monochromatic module 2; the grooves 2-1 on all the monochrome modules 2 are the same in shape and size. The outer walls of the die main body 3 and the transparent die cuvette 1 are both made by cutting acrylic plates.
The length, width and thickness of the die body 3 are 125mm, 85mm and 5mm, respectively. The internal length, internal width, internal height and wall thickness of the cuvette 1 of the transparent mould are 30mm, 10mm and 2mm, respectively. The interval between the outer walls of the adjacent small transparent mould pools 1 on the mould main body 3 is 6 mm. The opening width, the opening depth, the height and the wall thickness of the groove 2-1 are respectively 14mm, 10mm and 2 mm.
Example 2
This example differs from example 1 only in that in this example the internal length, internal width, internal height and wall thickness of the transparent mould cuvette 1 are 27mm, 8mm and 1.5mm, respectively; the opening width, the opening depth and the height of the groove 2-1 are respectively 11mm, 8mm and 8 mm.
Example 3
This example differs from example 1 only in that in this example the internal length, internal width, internal height and wall thickness of the transparent mould cuvette 1 are 33mm, 12mm and 2.5mm, respectively; the opening width, the opening depth and the height of the groove 2-1 are respectively 17mm, 12mm and 12 mm.
Example 4
The present embodiment is different from embodiment 1 only in that, in the present embodiment, the opening width of the groove 2-1 is 16 mm; the interval between the outer walls of the adjacent small transparent mould pools 1 on the mould main body 3 is 8 mm.
Application example 1: preference of 5dpf, 6dpf zebra fish larvae in blue and different color combinations
The multichannel instrument of example 1 was used to evaluate the preference of 5dpf, 6dpf zebra fish larvae in blue and different color combinations, with the following specific steps:
(1) selecting zebra fish: placing zebra fish of 5dpf and 6dpf (namely 5 days and 6 days after fertilization) under a dissecting microscope for observation, selecting the zebra fish which normally develop, and moving the zebra fish into 6 small transparent mould pools of a multi-channel appliance, wherein 5 pieces of 5dpf zebra fish are placed in each of No. 1-3 small transparent mould pools, and 5 pieces of 6dpf zebra fish are placed in each of No. 4-6 small transparent mould pools; then, using culture water to fix the volume of each transparent mould cuvette to 2 mL;
(2) installing a monochrome module: the blue module and the yellow module are respectively arranged at two ends of the No. 1 and No. 4 transparent mould small pools, the blue module and the red module are respectively arranged at two ends of the No. 2 and No. 5 transparent mould small pools, and the blue module and the green module are respectively arranged at two ends of the No. 3 and No. 6 transparent mould small pools;
(3) and (3) carrying out detection by a Viewpoint zebra fish behavioral instrument: putting a multi-channel instrument into a Viewpoint behavior detector, setting parameters of the Viewpoint behavior detector according to an operation manual, wherein the detection time is 20min, taking data of the last 10min, counting the total movement distance of the zebra fish in different color areas (the first 10min is the zebra fish adaptation period, so that the zebra fish adapts to the environment in the transparent mould cuvette), and calculating the ratio of the movement distance of the blue area of the zebra fish in each transparent mould cuvette according to the following formula:
wherein, the color A is blue, and the color B is another color in the same transparent mold cuvette.
(4) As a result: as shown in fig. 4A, 5dpf zebra fish larvae all showed a preference for blue in the three color combinations blue-yellow, blue-red, blue-green, and were statistically significant. When the blue and yellow are combined, 5dpf zebra fish juvenile fish has stronger preference for a blue area, and the movement distance of the blue area accounts for 73%. As shown in fig. 4B, 6dpf zebrafish larvae showed a preference for blue in both blue-yellow and blue-green color combinations and were statistically significant, with no statistically significant difference in the blue-red combinations. Wherein, the preference of 6dpf zebra fish juvenile fish to a blue area is stronger when the blue and yellow are combined, and the movement distance of the blue area accounts for 77%. Based on the above results, the blue-yellow two-color combination is preferably used in the color preference test of zebra fish juvenile fish due to its high stability and distinct difference in the results.
Application example 2: protective effects of edaravone on bisphenol af (bpaf) -induced zebra fish color cognitive impairment the protective effects of edaravone on bisphenol af (bpaf) -induced zebra fish color cognitive impairment were evaluated using the multichannel device of example 1, with the specific steps as follows:
(1) selecting zebra fish: observing zebra fish with 5dpf under a dissecting microscope, selecting the zebra fish with normal development, transferring the zebra fish into beakers, wherein 30 fish in each beaker are cultured, and the volume of fish culture water is 20mL, and the number of the zebra fish is 3;
(2) BPAF and edaravone treatment: the experiment was divided into 3 groups, one control group, one BPAF model group, and the other positive drug treatment group. Adding 20 mu L of BPAF mother solution with the concentration of 1mg/mL into a BPAF modeling group beaker until the exposure concentration is 1 mu g/mL; adding edaravone into a beaker of the positive drug treatment group to a final concentration of 5 mug/mL on the basis of BPAF molding; adding 20 mu L of DMSO solvent into the control group, and treating for 1 day;
(3) installing a monochrome module: transferring the zebra fish processed in the step (2) into a multi-channel appliance, and putting 5 fish into each transparent mould cuvette; a blue module and a yellow module are respectively arranged at two ends of each transparent mould cuvette;
(4) and (3) carrying out detection by a Viewpoint zebra fish behavioral instrument: putting the multichannel device into a Viewpoint behavior detector, setting parameters of the Viewpoint behavior detector according to an operation manual, wherein the detection time is 20min, taking data of 10min later, counting the total movement distance of the zebra fish in different color areas, and calculating the ratio of the movement distance of the blue area of the zebra fish in each transparent mould cuvette according to the following formula:
(5) as a result: as shown in fig. 5, the ratio of the moving distance of the zebra fish larvae in the normal group in the blue region was 82%, the ratio of the moving distance of the zebra fish larvae in the BPAF-treated group in the blue region was 21%, and the ratio of the moving distance of the zebra fish larvae in the positive drug edaravone-treated group in the blue region was 66%. The result shows that BPAF causes cognitive disorder of zebra fish juvenile fish on color, and edaravone can prevent cognitive disorder caused by BPAF.
Application example 3: the protective effect of edaravone on bisphenol af (bpaf) -induced zebrafish color cognitive impairment the present application example differs from application example 3 only in that the multichannel device of example 4 is employed in the present application example; and (4) placing the multichannel device into a Viewpoint behavior detector, setting parameters of the Viewpoint behavior detector according to an operation manual, standing for 10min, performing data acquisition for 8min, stopping data acquisition, exchanging the single-color modules at two ends of each transparent mold cuvette under the condition that the transparent mold cuvettes are not moved, continuing to perform data acquisition for 8min, and counting the total movement distance of the zebra fish in different color areas in the two data acquisition processes.
As a result: as shown in fig. 6, the proportion of zebra fish larvae in the blue region was 86% in the normal group, 24% in the blue region in the BPAF-treated group, and 72% in the blue region in the positive-drug edaravone-treated group. Compared with the application example 2, the ratio of the moving distance of each group of zebra fish juvenile fish in the blue area is improved, and the error caused by the zebra fish resting in the preferred color area can be reduced by regularly replacing the position of the single-color module in the data acquisition process.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (10)
1. A multichannel device for detecting the colour preference of zebra fish larvae, characterized by comprising at least one transparent mould cuvette (1) and at least two monochrome modules (2); all the monochrome modules (2) have at least two different colors; the single-color module (2) is provided with a groove (2-1) matched with the end part of the transparent mould cuvette (1), and the end part of the transparent mould cuvette (1) can be arranged in the groove (2-1) of the single-color module (2); the grooves (2-1) on all the single-color modules (2) are the same in shape and size.
2. The multichannel instrument as claimed in claim 1, characterized in that the number of monochromatic modules (2) is at least four, including at least one blue module, at least one yellow module, at least one red module and at least one green module.
3. A multichannel device as claimed in claim 1 or 2, characterized in that said transparent mould cuvette (1) is rectangular; the external width of the transparent mould cuvette (1) is not more than the opening width of the groove (2-1); and half of the external length of the small transparent mould pool (1) is not less than the opening depth of the groove (2-1).
4. The multichannel instrument as claimed in claim 3, wherein:
the external width of the transparent die cuvette (1) and the opening width of the groove (2-1) are both 11-17 mm; and/or
The external length of the transparent die cuvette (1) is 30-38 mm, and the opening depth of the groove (2-1) is 8-12 mm; and/or
The internal height of the transparent die cuvette (1) and the height of the groove are both 8-12 mm; and/or
The wall thickness of the transparent die cuvette (1) is 1.5-2.5 mm.
5. A method for detecting color preference of zebrafish larvae by using a multi-channel device according to any one of claims 1 to 4, comprising the steps of:
(1) putting the zebra fish juvenile fish into a small transparent mould pool, and adding water into the small transparent mould pool;
(2) installing a single-color module at each of two ends of each transparent mold cuvette respectively, so that the single-color modules at the two ends of each transparent mold cuvette have different colors, namely A color and B color;
(3) after the zebra fish juvenile fish adapts to the environment in the transparent mold small pool, data collection is carried out, and the total movement distance of the zebra fish juvenile fish in different color areas is counted;
(4) and judging the color preference of the zebra fish juvenile fish according to the total movement distance of the zebra fish juvenile fish in different color areas.
6. The method according to claim 5, wherein in the step (1), 4-6 zebrafish larvae are placed in each transparent mold cell; the zebra fish juvenile fish is 5-6 days after fertilization.
7. The method according to claim 5, wherein in the step (3), the data acquisition is performed in a continuous acquisition mode, and the data acquisition time is 10-15 min.
8. The method of claim 5, wherein in the step (3), the data acquisition adopts an interval acquisition mode, and the specific process is as follows: stopping data acquisition after 5-8 min of data acquisition, exchanging the single-color modules at two ends of the small transparent mould pool under the condition of not moving the small transparent mould pool, and then continuing to acquire data; and carrying out data acquisition for 2-4 times in total.
9. The method of claim 5, wherein the specific process of step (4) is as follows: calculating the ratio of the moving distances of the zebra fish juvenile in the color A area to the moving distances of the zebra fish juvenile in the color B area according to the total moving distances of the zebra fish juvenile in the color A area and the zebra fish juvenile in the color B area, and judging the preference of the zebra fish juvenile to the color A; the ratio of the movement distance of the color A region is calculated according to the following formula:
10. use of a multichannel device according to any of claims 1 to 4 or a method according to any of claims 5 to 9 for evaluating and screening a therapeutic agent for cognitive impairment.
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Denomination of invention: A multi-channel instrument for detecting color preferences of zebrafish juveniles and its usage and application Granted publication date: 20221118 Pledgee: Industrial and Commercial Bank of China Limited Hangzhou Jiangnan Branch Pledgor: HANGZHOU HUNTER BIOTECHNOLOGY Co.,Ltd. Registration number: Y2024980009699 |
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