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Method Article

Phenotypic Profiling of MPTP-Induced Parkinsonian-like Behavioral Phenotypes in Zebrafish Larvae Based on Behavioral Experiment

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DOI:

10.3791/70041

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May 15th, 2026

 ,  ,  ,  ,  ,  ,  , 
1Department of Medicinal Chemistry, State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, Key Laboratory of Chemical Biology (Ministry of Education), NMPA Key Laboratory for Technology Research and Evaluation of Drug Products, School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, 2College of Information Engineering, Ningde Normal University, 3Digital Fujian Research Institute of Big Data for Elderly Rehabilitation and Nursing, 4Department of Neurology, Qilu Hospital Cheeloo College of Medicine, Shandong University, 5Key Laboratory of Bioactive Materials, Ministry of Education, Nankai University, 6Jinan Microecological Biomedicine Shandong Laboratory

In This Article

Summary

This protocol establishes a behavioral phenotyping paradigm in MPTP-treated zebrafish larvae to evaluate Parkinson's disease-like motor and non-motor deficits.

Abstract

This article introduces a unique, multiparametric behavioral phenotyping protocol for MPTP-induced Parkinson's disease (PD) in larval zebrafish. Chemical induction of PD-like pathology in zebrafish provides a high-throughput, genetically tractable system, yet most studies have focused narrowly on basic locomotor assays or simple light-dark transitions. Such limited approaches fail to capture the broader spectrum of PD-related dysfunctions, ranging from bradykinesia and sensorimotor deficits to anxiety-like behaviors and sleep disturbances. To address this gap, this study developed an integrated battery of 6 complementary assays: locomotor test to assess general locomotor and exploratory drive; thigmotaxis assessment to detect anxiety-related edge-preference behaviors; startle response test to record bradykinesia and impaired motor reactivity; light-dark challenge assays to evaluate risk-taking and stress responsiveness; photomotor response paradigms (including strobe light stimulation) to quantify sensorimotor and reflexive movement; and sleep-wake regulation monitoring to measure sleep-wake behavior. Larval zebrafish are exposed to sublethal concentrations of MPTP from 1 day post-fertilization (dpf) to 5 dpf, with behavioral assessments initiated from 4 to 6 dpf. High-resolution automated tracking generates several behavioral endpoints per assay. This approach uncovers previously undetected PD-like alterations, such as disorderly sleep architecture, impaired habituation learning, sensorimotor and reflexive movement, reflecting early nonmotor symptoms. By combining these assays, researchers obtain a comprehensive behavioral symptom that more closely mirrors the complexity of human PD. This protocol expands the paradigm for studying PD pathogenesis, screening candidate therapeutics, and exploring environmental modulators. It exemplifies the collection's goal of presenting innovative animal models that enhance translational relevance through robust, multifaceted phenotyping.

Introduction

Parkinson's disease (PD) is the second most prevalent neurodegenerative disorder after Alzheimer's disease1. Its prevalence and the number of individuals affected have been steadily increasing in recent years, presenting a significant public health challenge worldwide2. The incidence rate of PD notably escalates with age3,4. Clinically, PD manifests through both motor symptoms -- such as dyskinesia, bradykinesia, resting tremor, and postural instability -- and a variety of non-motor symptoms, including cognitive impairment, anxiety, and sleep disturbances5. Currently, the diagnosis of PD primarily depends on the presence of motor symptoms6; however, by the time these symptoms become evident, over 60% of the dopamine neurons in the substantia nigra have already been lost7. Although non-motor symptoms frequently appear earlier, the absence of specific and objective assessment criteria complicates early diagnosis and intervention8. Presently, the primary treatment for PD involves the administration of levodopa9, which is converted into dopamine in the brain, thereby replenishing dopamine levels in the striatum. While this treatment can temporarily alleviate symptoms, it does not halt the progression of the disease10. Consequently, further research into the pathogenesis of PD and drug screening is essential.

Building appropriate animal models is crucial for the research on the pathogenesis of PD and anti-PD drug screening. Now, there are three main models for PD studies: chemical induction model11, gene editing model12, and human xenotransplantation model13. Commonly used model animals include rodents, such as mice14, and non-human primates like monkeys15. Recently, zebrafish (Danio rerio) have gained increasing attention in PD studies for their advantages such as small size, rapid growth, high genetic homology with humans, and transparent embryos16. Using neurotoxin MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) to induce dopamine neuron damage17,18, is a classical method to construct a PD model. Zebrafish exposed to sublethal MPTP concentrations exhibit various PD-like behavioral phenotypes19.

However, most zebrafish PD studies nowadays mainly concentrate on basic locomotor behaviors or simple light-dark transitions16,20, and single-dimensional behavioral assays are difficult to fully reflect the complex behavioral characteristics of PD, especially the behavioral changes in non-motor function. Considering PD is a disease that involves multiple neurobehavioral systems beyond motor control21,22,23, a multiparametric behavioral phenotype evaluation method is urgently needed to capture its multisystem phenotypes24.

Accordingly, this study builds a multiple, parametric, and rapid behavioral phenotyping assay protocol to more comprehensively describe the PD-like behavioral phenotypes in MPTP-treated zebrafish larvae. A sublethal dose of MPTP (50 µM)25 was applied to larvae from 1 to 5 days post-fertilization (dpf)16,20,26. From 4 to 6 dpf16,20,24, we conducted 6 behavioral tests: sleep-wake regulation monitoring, photomotor response paradigms, thigmotaxis assessment, locomotor assessment, startle response test, and light-dark challenge assays. These behavioral assessments cover both motor and non-motor aspects, such as movement deficits, sensorimotor, habituation learning, risk-taking, stress responsiveness, and sleep-wake rhythm.

Utilizing the Zebrabox high-throughput video tracking system and quantitative analysis method, we extracted and quantified various behavioral parameters and compared the behavioral differences between the MPTP-treated group and the wild-type (WT) group by statistics. Compared with existing zebrafish PD assays, this multiparametric behavioral battery captures motor and non-motor phenotypes within a standardized framework. The automated tracking system ensures objective behavioral data acquisition and high reproducibility. In addition, the small body size of larvae allows individual animals to be placed in multi-well plates, enabling high-throughput assessment. The above advantages are beneficial for deeply analyzing the pathological mechanism of PD, screening the candidate drugs or compounds, and exploring the influence of environmental factors on the occurrence or development of PD.

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Protocol

This study involving zebrafish was conducted in compliance with institutional guidelines for animal care and use, and was approved by the Ethical Committee of the School of Pharmaceutical Sciences, Shandong University (No.YXDW2025-0056). The reagents and the equipment used are listed in the Table of Materials.

1. MPTP Preparation

  1. Prepare 1× E3 medium by diluting 100× E3 stock solution 1:100 with ultrapure water (e.g., mix 5 mL of 100× E3 with 495 mL of ultrapure water to obtain 500 mL of 1× E3 medium).
    NOTE: Prepare E3 medium freshly on the day of use.
  2. Prepare a 1 mM MPTP stock solution by adding 5.0 mg of MPTP powder to a 50 mL centrifuge tube and adding E3 medium to a final volume of 25 mL (use the centrifuge tube as a mixing container, but no centrifugation is required). Label the solution as 1 mM MPTP stock solution. Store the stock solution in the dark.
  3. Preparation of 50 µM MPTP solution: transfer 2.5 mL of the 1 mM MPTP stock solution into a clean container and dilute with 47.5 mL of E3 medium to obtain a final volume of 50 mL at a final concentration of 50 µM MPTP. The remaining part of the stock solution continued to be stored in the dark.
    CAUTION: MPTP is a potent neurotoxin and must be handled with extreme caution. Always wear appropriate personal protective equipment (PPE), including gloves, a laboratory coat, and protective eyewear. Disposal of MPTP waste according to institutional biosafety guidelines.

2. Zebrafish embryos preparation and grouping

  1. Select sexually mature male and female adult zebrafish and perform controlled breeding using breeding tanks with removable dividers. At 8:00 pm, place male and female zebrafish separately in the breeding tanks at a ratio of 2:3, and separate them with dividers.
  2. On the following day, remove dividers at 8:00 am to allow mating, thereby initiating spawning. Collect embryos after 1 h, and wash them three times with clean system water.
  3. House embryos in 100 mm Petri dishes with fresh E3 medium, and incubate them at 28 °C under a 14 h light / 10 h dark cycle (lights on from 08:00 am to 10:00 pm).
    NOTE: Do not exceed 50 embryos per dish with 20 mL medium to ensure normal development.
  4. At 1 dpf, carefully remove the unfertilized, malformed, and abnormal embryos using a Pasteur Pipette, and replace the solution containing zebrafish embryos. For the WT group, replace the existing medium with fresh E3 embryo medium. For the MPTP-treated group, replace the medium with 50 µM MPTP solution (Figure 1A), then return both groups to the incubator.
  5. For both groups, replace the solution and remove any dead or unhealthy embryos every 24 h. For the MPTP-treated group, replace the MPTP solution daily from the morning of 1 dpf until the morning of 4 dpf (Figure 1A). Then, transition the MPTP-treated group to E3 medium beginning on the morning of 5 dpf until the end of the experiment.
    NOTE: Replace the solution gently using a Pasteur Pipette. Aspirate the medium slowly from the edge of the Petri dish and add fresh solution along the wall of the Petri dish to minimize disturbance to the embryos.

3. Sleep - wake regulation monitoring

  1. At 4 dpf, select zebrafish larvae from each group at 6:00 pm, and transfer them to a 96-well plate using a modified 3 mL Pasteur Pipette with the tip cut off, one larva per well (Figure 1B). The first four rows represent the WT groups, and the last four rows represent the MPTP groups (48 zebrafish in each group).
    NOTE: To avoid stimulating or damaging zebrafish, modify Pasteur Pipettes by cutting the tip (10 mm) to provide a wide bore larger than the size of 4 dpf zebrafish larvae (body length exceeds 3.7 mm27,28).
  2. Observe each well with a stereo microscope to ensure the appearance of each zebrafish larva is healthy, the fins are open and regularly swung, the body is upright, and the swim bladder is inflated29. After ensuring that the zebrafish in each well are in a healthy state, fill each well with E3 medium.
    NOTE: Fill each well with E3 medium to near bulging, ensuring the surface tension is maintained. Carefully add the liquid to prevent bubbles and cross-well overflow.
  3. After transferring the fish and liquid into a 96-well plate, place it in the Zebrabox chamber to acclimate for 30 min prior to the sleep-wake behavioral test.
    NOTE: Before placing the 96-well plate in the Zebrabox chamber, clean up the bottom of the plate carefully to avoid dirt affecting the video tracking results.
  4. Open the Zebralab video tracking software, select the Quantization mode, and ensure that the 96-well plate is positioned centrally on the bottom of the Zebrabox chamber.
  5. Set the light adjustment to turn off the light at 10: 00 pm and turn on the light at 8: 00 am, which is consistent with the light conditions when the zebrafish is reared. During the daytime, white light is generally around 1000 lux. Set the software parameters as follows29: detection threshold, 40; burst, 25 (threshold for on movement); freeze (threshold for on movement), 4; bin size, 60 s; total monitoring time, 52 h (8: 00 pm on the first day to 00: 00 pm on the third day). Start the sleep-wake procedure at 8: 00 pm.
    NOTE: Once the procedure is started, prohibit any noise and vibration from the outside throughout the experiment, so as not to affect the normal sleep-wake rhythm of zebrafish. To prevent water evaporation during long-term monitoring, cover the 96-well plate with transparent plastic wrap.
  6. After 52 h, stop the sleep-wake experiment, and save the data file. Remove the 96-well plate from the ZebraBox.
    NOTE: Using MATLAB and Excel, necessary parameters such as number of sleep bouts (sleep bout is defined as periods of inactivity lasting at least 1 min)29, average sleep bout length (average duration of sleep bouts), total sleep duration (all duration of sleep bouts over the monitoring period), sleep latency (time from a light transition to the onset of the first sleep bout), average activity (defined as locomotor activity during all the monitoring period), and waking activity (defined as locomotor activity during only the awake period, excluding the sleep bouts) were extracted and analyzed from the data file.
    1. Use a Student's t-test to compare the differences between the WT group and the MPTP-treated group. Additionally, plot the sleep curve, waking activity curve, and average activity curve of zebrafish over the 52 h using MATLAB software.

4. Photomotor response paradigms

NOTE: Photomotor response paradigms (including strobe light) are high-throughput behavioral tests in 5 dpf larvae to quantify sensorimotor and reflexive locomotor changes in response to a sudden transition of light. This test was conducted using a 96-well plate, including 2 paradigms: (1) a light transition from 1 min of darkness to 1 min of constant light (1000 lx), and (2) a strobe light response involving a transition from 1 min of darkness to 1 min of strobing light at 10 Hz.

  1. Place the 5 dpf larvae in a 96-well plate (1 larva per well) (Figure 1B), and fill each well with E3 medium. Divide a total of 96 larvae into the WT group and the MPTP group on average.
  2. Dark-constant light paradigm:
    1. Place the 96-well plate with 5 dpf larvae into the Zebrabox and allow larvae to acclimate for 5 min in the dark before the assay.
    2. Open the Tracking-online analysis mode.
    3. Set the light transition program as follows: 0-60 s: Dark, 60-120s: Constant light (1000 lx).
    4. Start the protocol, collect related motion parameters every 6 s throughout the 120 s assay.
    5. Stop the protocol at 120th s, export the data file. Place the larvae in full darkness for 5 min to acclimate the environment, preparing them for the following dark-strobe light assay.
  3. Dark-strobe light paradigm:
    1. Set the light transition program as follows: 0-60 s: Dark, 60-120s: strobe light (1000 lx) at 10 Hz.
      NOTE: A 10 Hz strobe corresponds to 10 light pulses per second. In ZebraLab, implement this by alternating the light intensity from 0 to 1000 lx every 100 ms.
    2. Upon completion of the 5 min dark adaptation, start the dark-to-strobe light assay. Collect motion parameters every 6 s throughout the 120 s assay.
  4. Data analysis: Once the assay ends, export the data file. Use MATLAB software to extract the swimming distance every 6 s. Plot the total distance (mean value per 6 s) swum in the 2 min period for both assays. Calculate freeze index by subtracting the total distance traveled in 1 min in the dark from the total distance traveled in the light30.
    NOTE: Freeze Index < 0 indicates freezing behavior; Freeze index > 0 indicates escape; Freeze index = 0 means no reaction. Use a Student's t-test and two-way ANOVA to compare the differences between the WT group and the MPTP-treated group. A total of 48 zebrafish larvae (n = 48) were used for each group.

5. Assessment of thigmotaxis

  1. Select two 6-well plates, fill each well with 4 mL E3 medium.
  2. At 5 dpf, zebrafish larvae from the WT group and MPTP-treated group were collected and put in two 6-well plates (one is the WT group, and the other is the MPTP-treated group, 1 larva per well) (Figure 1C).
    NOTE: Ensure the initial position of the larvae is in the center of the wells.
  3. Before the thigmotaxis assessment, the well plates should be placed in the appropriate position of Zebrabox 15 min in advance to accommodate.
  4. Data collection and analysis: Using the Draw Area function, draw two concentric circles in the observation area of each well (the radius of the center circle accounts for 1 / 2 of the radius of the hole) (Figure 1C), and the remaining parameters are set to be the same as locomotor assessment. Start the protocol for 5 min thigmotaxis assessment, and do a parallel experiment.
    NOTE: After the end of the program, Zebralab software will generate the trajectory of zebrafish larvae within 5 min and various related motion parameters. In the data file, an1 represents the whole area of each well, and an2 represents the inner zone of each well. Thus, total dis(an1) and total dur(an1) denote the total swimming distance and total time spent in the entire well, while total dis(an2) and total dur(an2) represent the distance and time spent in the inner zone, respectively.
    1. Calculate the thigmotaxis with the following formulas:
      Calculate the percent of distance swum in the outer zone:
      Thigmotaxis calculation formula; equation showing behavioral analysis method in research context.
      Calculate the percent of time spent in the outer zone:
      Thigmotaxis equation, ΣFx=0, describing behavioral analysis, includes parameters total dur, an1, an2.
    2. Employ a Student's t-test to compare the differences of thigmotaxis between the WT group and the MPTP-treated group. A total of 24 zebrafish larvae (n = 24) were used for each group.

6. Locomotor assessment

  1. At 5 dpf, select zebrafish larvae from the WT group and MPTP-treated group, and respectively transferred to two 12-well plates (one is the WT group, and the other is the MPTP-treated group, 1 larva per well, with 2 mL E3 medium) (Figure 1D).
    NOTE: Ensure the sizes of zebrafish larvae are similar, and larvae selected should have open fins, upright posture, good expansion of the swim bladder, and regular movement.
  2. Before the locomotor assessment, place the well plate in the test position of the Zebrabox 15 min in advance to adapt.
    NOTE: Ensure the light conditions during adaptation are consistent with the light conditions during the test.
  3. After a 15 min acclimation period, select the tracking mode-online analysis, record the locomotor activity of each larva for 5 min using an automated computerized video-tracking system. Using Zebralab software to collect movement tracks and a variety of related locomotor parameters every 60 s, export the complete data file, and use MATLAB to analyze the total distance and average speed of each larva in 5 min.
    1. Use a Student's t-test and two-way ANOVA to compare the differences between the WT group and the MPTP-treated group. A total of 24 zebrafish larvae (n = 24) were used for each group.

7. Startle response test

NOTE: This test measures the visual startle response (VSR), also known as dark-flash induced startle, elicited by sudden transitions to darkness. This startle response test assesses a photic-evoked visual startle induced by abrupt, millisecond-scale dark flashes. Unlike the dark-strobe light paradigm, which involves repeated light pulses to evaluate sustained behavioral responses, this assay focuses on rapid, reflexive motor reactions to an acute visual stimulus.

  1. Preparations: at 5 dpf, select zebrafish larvae from the WT group and the MPTP-treated group, and transfer them respectively to two 12-well plates (one is the WT group, and the other is the MPTP-treated group, 1 larva per well, with 2 mL E3 medium) (Figure 1D).
  2. Before the startle response test, place the well plate in the test position of the Zebrabox 5 min in advance to adapt.
  3. Set the stimulation parameters: open Zebralab software, run the Tracking-online analysis mode, open the Light-triggering interface, select Use one of the 3 triggering methods below, enable Enhanced stimuli, set to When session is started; in the transition schedule, the dark flash rhythm is defined as follows ( the program starts to trigger after 21 s ): Transition 1: 5 ms, 100% light intensity; transition 2: 2890 ms, 100% light intensity; transition 3: 5 ms, 0% light intensity; transition 4: 100 ms, 0% light intensity; tick Repeat list of transitions to ensure periodic flashing.
  4. Start protocol: ensure that the Numeriscope video recording function is turned on, high-throughput automated tracking will generate a multi-dimensional dataset encompassing 30 behavioral endpoints in this 90 s test.
  5. Data analysis: After completion of the startle response assay, export the raw data files and import them into MATLAB software for analysis. Calculate the swimming distance of each larva in 3 s intervals, along with the average swimming speed.
    1. Identify the peak movement distance, peak velocity, or response latency following the flash stimulus for each group. Use a Student's t-test and two-way ANOVA to compare the differences between the WT group and the MPTP-treated group. A total of 24 zebrafish larvae (n = 24) were used for each group.

8. Light-dark challenge assay

  1. At 5 dpf, transfer zebrafish larvae individually into two 24-well plates (Figure 1E), with one larva per well and 1 mL of E3 medium per well. Each plate contained 24 larvae, and the two 24-well plates were evenly divided into the WT group and the MPTP-treated group (n = 24 per group).
  2. Before testing, place the 24-well plate into the designated testing area inside the Zebrabox for a 5 min light acclimatization period to adapt the testing environment. Activate the Tracking-Online Analysis mode. Configure the light program as follows: 6 min of constant light followed by 4 min of complete darkness (Figure 1E). The total testing time is 10 min, and the interval time for collecting data points is 60 s.
    NOTE: The global 6 min of light is for acclimatization and habituation, ensuring stable baseline locomotor and special light-dark responses31,32.
  3. On the 10th min, stop behavioral recording and save data. Carefully remove the plate from the Zebrabox.
  4. For each larva, extract the swimming distance at each 60 s interval. Calculate the distance traveled during the light phase (0th-6th min), the dark phase (7th-10th min), and the total 10-min period. Quantify the thigmotaxis behavior and habituation learning performance in the dark phase.
    NOTE: Habituation learning is defined as the reduction of a behavioral response when an animal is exposed to a continuous stimulus/ environment33,34, and it was assessed by statistically comparing the mean total distance moved (mm) during the 1 st min and the 4th min following the light-to-dark transition. Use a Student's t-test and two-way ANOVA to compare the differences between the WT group and the MPTP-treated group. A total of 24 zebrafish larvae (n = 24) were used for each group.

9. Disposal of MPTP waste

NOTE: After each behavioral experiment, spray and wipe MPTP-contacted surfaces (e.g., lab benches and the ZebraBox) with freshly prepared sodium hypochlorite (bleach; ≥10% v/v).

  1. Collect the liquid waste (including remaining exposure solutions and rinsing media) and solid waste (e.g., Pasteur pipettes, gloves, Petri dishes, and multi-well plates) of MPTP. Clearly label them as hazardous neurotoxic waste.
  2. Treat contaminated liquid waste and solid materials with freshly prepared sodium hypochlorite (bleach; ≥10% v/v) for at least 24 h to ensure decontamination, followed by disposal in accordance with institutional regulations.
  3. Euthanize MPTP-treated zebrafish larvae by immersing in a diluted sodium hypochlorite solution (e.g., 1:5 ratio of 6% bleach: water) for 5 min35. Confirmed death by lack of movement and heartbeat. Treat the decontaminated larvae and media as hazardous biological waste for institutional disposal (following institutionally approved protocols).

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Results

Sleep-wake regulation monitoring
The 52-h sleep-wake regulation monitoring began at 4 dpf and was conducted using a 96-well plate. Zebrafish in MPTP-treated showed significant disruptions in sleep structure compared to the WT group. Representative sleep curves, wake activity plots, and averaged activity traces are shown in Figure 2. Parameters of sleep-wake regulation analysis in zebrafish larvae are shown in Figure 3. Total sleep time was s...

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Discussion

This study developed and verified a comprehensive protocol to evaluate and analyze MPTP-induced PD-like behavioral phenotype. This protocol integrated 6 different tests, locomotor assessment, thigmotaxis assay, startle response test, photomotor response paradigms (including strobe light stimulation), light-dark challenge assay, and sleep-wake regulation monitoring, to evaluate and depict the motor and non-motor defects of PD. The results indicated that compared with the WT group, MPTP-treated zebrafish larvae showed moto...

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Disclosures

The authors declare no conflicts of interest.

Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (No. 21906095), Shandong University Young Scholar Future Program Sponsorship, the Shandong Natural Science Foundation (No. ZR2019BB018), the Fundamental Research Funds of Shandong University (No. 2017GN0030), the China Postdoctoral Science Foundation (No. 2018M640640). It is also supported by both the 2025 Open Research Fund from the Digital Fujian Institute of Big Data for Elderly Rehabilitation and Nursing (Grant No. BDRI202502) and the 2025 Open Research Fund from the Key Laboratory of Bioactive Materials, Ministry of Education, Nankai University (Grant No. SWHX-202503). Research Project of Jinan Microecological Biomedicine Shandong Laboratory (Grant No.JNL-2023002KF). The authors would like to thank the Laboratory Animal Center, Qilu Hospital of Shandong University, and Dr. Sun Chengxi from the Department of Clinical Laboratory, Qilu Hospital of Shandong University, for their assistance during the experiment.

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
100× E3 stock solutionShanghai Feixi BiotechnologyN/Acontaining 34.8 g NaCl, 1.6 g KCl, 5.8 g CaCl2·2H2O, and 9.78 g MgCl2·6H2O per liter
12-Well Cell Culture PlateNEST Biotechnology712011Non-Treated, Non-Pyrogenic, Sterile, Polystyrene.Used for zebrafish larval behavioral assays
24-Well Cell Culture PlateNEST Biotechnology702011Non-Treated, Non-Pyrogenic, Sterile, Polystyrene.Used for zebrafish larval behavioral assays
3 mL Pasteur PipetteBiologix30-0238A1
6-Well Cell Culture PlateNEST Biotechnology703011Non-Treated, Non-Pyrogenic, Sterile, Polystyrene.Used for zebrafish larval behavioral assays
96-Well Cell Culture PlateNEST Biotechnology701011Non-Treated, Non-Pyrogenic, Sterile, Polystyrene.Used for zebrafish larval behavioral assays
MATLAB R2020aMathWorks, USA N/AMATLAB R2020a, Campus-wide license obtained from Shandong University software portal (softms.sdu.edu.cn).Used for statistical analysis of zebrafish larval behavioral data.
Microsoft ExcelMicrosoftN/AMicrosoft Excel (part of Microsoft Office), version installed via Shandong University licensed software portal (softms.sdu.edu.cn).
MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) Beyotime411252815 mg
Petri DishNEST Biotechnology704202100 mm, sterile, easy grip handle, TC-Treated
Sodium hypochlorite solutionMacklinS817439stock household bleach solution(6-14% active chlorine basis), diluted to ≥10% v/v for surface and waste decontamination; also diluted 1:5 (6% bleach: water) for euthanasia of MPTP-treated zebrafish larvae.
Stereo MicroscopeOlympusSZX2 series
Zebrabox High-Resolution Video Tracking System Viewpoint, France N/A
Zebralab Software Viewpoint, France N/AVersion (3,52,3,87)

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