Name: In Vivo Phenotyping of Dopaminergic Neurodegeneration in Zebrafish Larvae Using Behavioral Analysis and High-Content Imaging (Video) | JoVE Journal
Uploaded: 2026-06-16T13:40:00.000+00:00
Duration: 8 min 13 s
Description: Discover a high-throughput phenotyping pipeline using transgenic zebrafish to study dopaminergic neuron loss in Parkinson's disease. This method integrates chemogenetic ablation with behavioral analysis and high-content imaging for rapid drug screening.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

playerTrialDesktop

We thank Dr. Su Guo (University of California, San Francisco) for providing the transgenic zebrafish, and Dr. Daeseok Eom, Dr. Michael Parsons, and Eddie Hallo (University of California, Irvine) for providing wild-type embryos and guidance on zebrafish husbandry and fish facility maintenance. We also thank the staff members of the Robert A. Mah Molecular Innovation Center and the Falling Leaves Innovation Building (University of California, Irvine) for providing infrastructure for the aquatics facility.

<ol>
	<li>Haynes, E. M., Ulland, T. K., Eliceiri, K. W. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+of+discovery:+the+role+of+imaging+established+and+emerging+non-mammalian+models+in+neuroscience.">A model of discovery: the role of imaging established and emerging non-mammalian models in neuroscience.</a> Front Mol Neurosci. 15, 867010 (2022).</li><li>Yang, W., Chen, X., Li, S., Li, X. J. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetically+modified+large+animal+models+for+investigating+neurodegenerative+diseases.">Genetically modified large animal models for investigating neurodegenerative diseases.</a> Cell Biosci. 11 (1), 218 (2021).</li><li>Dodiya, R., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish-based+Parkinson&#8217;s+disease+models:+unveiling+genetic+mechanisms+and+therapeutic+pathways.">Zebrafish-based Parkinson&#8217;s disease models: unveiling genetic mechanisms and therapeutic pathways.</a> CNS Neurol Disord Drug Targets. 24 (12), 900-920 (2025).</li><li>Wasel, O., Freeman, J. L. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+and+genetic+zebrafish+models+to+define+mechanisms+of+and+treatments+for+dopaminergic+neurodegeneration.">Chemical and genetic zebrafish models to define mechanisms of and treatments for dopaminergic neurodegeneration.</a> Int J Mol Sci. 21 (17), (2020).</li><li>Kozol, R. A., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+over+form:+modeling+groups+of+inherited+neurological+conditions+in+zebrafish.">Function over form: modeling groups of inherited neurological conditions in zebrafish.</a> Front Mol Neurosci. 9, 55 (2016).</li><li>Jang, H., Kim, S. H., Koh, Y., Yoon, K. J. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engineering+brain+organoids:+toward+mature+neural+circuitry+with+an+intact+cytoarchitecture.">Engineering brain organoids: toward mature neural circuitry with an intact cytoarchitecture.</a> Int J Stem Cells. 15 (1), 41-59 (2022).</li><li>Vandamme, T. F. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+rodents+as+models+of+human+diseases.">Use of rodents as models of human diseases.</a> J Pharm Bioallied Sci. 6 (1), 2-9 (2014).</li><li>Malik, N., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Compounds+with+species+and+cell+type+specific+toxicity+identified+in+a+2000+compound+drug+screen+of+neural+stem+cells+and+rat+mixed+cortical+neurons.">Compounds with species and cell type specific toxicity identified in a 2000 compound drug screen of neural stem cells and rat mixed cortical neurons.</a> Neurotoxicology. 45, 192-200 (2014).</li><li>Hasani, H., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-brain+imaging+of+freely-moving+zebrafish.">Whole-brain imaging of freely-moving zebrafish.</a> Front Neurosci. 17, 1127574 (2023).</li><li>Mattern, K., von Trotha, J. W., Erfle, P., K&#x00F6;ster, R. W., Dietzel, A. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NeuroExaminer:+an+all-glass+microfluidic+device+for+whole-brain+in+vivo+imaging+in+zebrafish.">NeuroExaminer: an all-glass microfluidic device for whole-brain in vivo imaging in zebrafish.</a> Commun Biol. 3 (1), 311 (2020).</li><li>Panier, T., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+functional+imaging+of+multiple+brain+regions+in+intact+zebrafish+larvae+using+selective+plane+illumination+microscopy.">Fast functional imaging of multiple brain regions in intact zebrafish larvae using selective plane illumination microscopy.</a> Front Neural Circuits. 7, 65 (2013).</li><li>Noda, S., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging-related+motor+function+and+dopaminergic+neuronal+loss+in+C57BL/6+mice.">Aging-related motor function and dopaminergic neuronal loss in C57BL/6 mice.</a> Mol Brain. 13 (1), 46 (2020).</li><li>Godoy, R., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemogenetic+ablation+of+dopaminergic+neurons+leads+to+transient+locomotor+impairments+in+zebrafish+larvae.">Chemogenetic ablation of dopaminergic neurons leads to transient locomotor impairments in zebrafish larvae.</a> J Neurochem. 135 (2), 249-260 (2015).</li><li>Schallert, T., Tillerson, J. L. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intervention+strategies+for+degeneration+of+dopamine+neurons+in+parkinsonism:+optimizing+behavioral+assessment+of+outcome.">Intervention strategies for degeneration of dopamine neurons in parkinsonism: optimizing behavioral assessment of outcome.</a> Cent Nerv Syst Dis. , 131-151 (2000).</li><li>Westerfield, M. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The zebrafish book: a guide for the laboratory use of zebrafish (Danio rerio). , (2007).</li><li>Avdesh, A., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regular+care+and+maintenance+of+a+zebrafish+(Danio+rerio)+laboratory:+an+introduction.">Regular care and maintenance of a zebrafish (Danio rerio) laboratory: an introduction.</a> J Vis Exp. (69), e4196 (2012).</li><li>Nipu, N., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methylene+blue+at+recommended+concentrations+alters+metabolism+in+early+zebrafish+development.">Methylene blue at recommended concentrations alters metabolism in early zebrafish development.</a> Commun Biol. 8 (1), 120 (2025).</li><li>Ohnesorge, N., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+the+effect+of+tricaine+(MS-222)-induced+anesthesia+on+brain-wide+neuronal+activity+of+zebrafish+larvae.">Assessment of the effect of tricaine (MS-222)-induced anesthesia on brain-wide neuronal activity of zebrafish larvae.</a> Front Neurosci. 18, 1456322 (2024).</li><li>Schroeder, A. B., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ImageJ+ecosystem:+open-source+software+for+image+visualization,+processing,+and+analysis.">The ImageJ ecosystem: open-source software for image visualization, processing, and analysis.</a> Protein Sci. 30 (1), 234-249 (2021).</li><li>Stirling, D. R., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CellProfiler+4:+improvements+in+speed,+utility+and+usability.">CellProfiler 4: improvements in speed, utility and usability.</a> BMC Bioinformatics. 22 (1), 433 (2021).</li><li>Kim, G. H. J., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+zebrafish+screen+reveals+renin-angiotensin+system+inhibitors+as+neuroprotective+via+mitochondrial+restoration+in+dopamine+neurons.">A zebrafish screen reveals renin-angiotensin system inhibitors as neuroprotective via mitochondrial restoration in dopamine neurons.</a> eLife. 10, (2021).</li><li>Kim, G. H. J., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+dopamine+neuron+imaging-based+small+molecule+screen+identifies+novel+neuroprotective+compounds+and+targets.">In vivo dopamine neuron imaging-based small molecule screen identifies novel neuroprotective compounds and targets.</a> Front Pharmacol. 13, 837756 (2022).</li><li>Aponte, P. M., Gutierrez-Reinoso, M. A., Garcia-Herreros, M. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bridging+the+gap:+animal+models+in+next-generation+reproductive+technologies+for+male+fertility+preservation.">Bridging the gap: animal models in next-generation reproductive technologies for male fertility preservation.</a> Life (Basel). 14 (1), (2023).</li><li>Fen, W., et al. <a target="_blank" href="https://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+for+cell-based+screening+methods+in+drug+discovery.">A review for cell-based screening methods in drug discovery.</a> Biophys Rep. 7 (6), 504-516 (2021).</li></ol>

The authors declare no financial conflicts of interest.

This protocol offers a scalable, in vivo pipeline for quantifying DA neuronal loss and screening neuroprotective compounds using the NTR&#8211;MTZ chemogenetic system in transgenic zebrafish. The standardized, plate-based workflow was specifically developed for undergraduate research teams, where operator turnover is high and investigator experience varies between academic terms. Each step of the workflow has been structured to prioritize reproducibility and ease of training, enabling consistent execution across multiple...

Drug discovery research in neurodegeneration faces a significant efficiency bottleneck due to the high cost and low throughput of traditional mammalian models1,2, requiring careful experimental planning and the use of a large workforce. The overall goal of this method is to provide a standardized, high-throughput approach using the Tg(th:Gal4; UAS:NTR-mCherry) transgenic zebrafish model in which nitroreductase-mCherry is specifically expressed in dopaminergic (DA) neurons to study DA neuron loss and locomotion deficits. The rationale behind the use of this technique stems from the need for rigorous, scalable, and economically viable in vivo models for neurodegeneration research3,4,5. While in vitro cell cultures allow for rapid drug screening, they lack the intact neural circuitry and whole-organism context essential for translational relevance6. In contrast, rodent models provide excellent physiological context but are expensive and time-consuming for initial high-throughput screening of neuroprotective compounds7,8.
Zebrafish possess rapid larval development, optical transparency that enables in vivo whole-brain imaging, and strong genetic conservation of disease-relevant pathways, particularly related to neurodegeneration9,10,11. The protocol specifically integrates standardized husbandry, chemogenetic ablation via metronidazole (MTZ), and high-content imaging paired with functional behavioral tracking. Measuring locomotor activity directly provides a robust behavioral correlate to DA neuronal loss, as dopamine neuron loss is tied to behavioral abnormalities12,13,14. The standardized, plate-based format minimizes experimental variance across operators, and the high throughput of the zebrafish larval system enables large sample sizes, together providing sufficient statistical power to detect neuroprotective effects. This makes the protocol well suited for applications requiring scalable and reproducible screening, including settings where investigator experience may vary.

<table><tbody><tr><td>Adult zebrafish diet, 0.5 mm pellets</td><td>Zeigler</td><td>AH271</td><td>Used for feeding adult zebrafish during routine husbandry</td></tr><tr><td>Agarose, low melting point (LMA)</td><td>Sigma-Aldrich</td><td>A9414-10G</td><td>Used for embedding larvae in wells for stable positioning during imaging</td></tr><tr><td>Aluminum foil</td><td>ULINE</td><td>S-20197</td><td>Used to protect light-sensitive reagents such as MTZ during preparation and incubation</td></tr><tr><td>CellProfiler</td><td>Broad Institute</td><td>RRID: SCR_007358</td><td>Used for optional automated fluorescence image analysis</td></tr><tr><td>Conical tubes, 15 mL</td><td>Corning</td><td>352097</td><td>Used for reagent preparation and sample handling</td></tr><tr><td>Conical tubes, 50 mL</td><td>Corning</td><td>352070</td><td>Used for reagent preparation and solution storage</td></tr><tr><td>Crossing tanks (1.0 L)</td><td>Aquaneering</td><td>ZHCT100</td><td>Used for controlled zebrafish breeding and embryo collection</td></tr><tr><td>Danio rerio (AB wild type)</td><td>&amp;mdash;</td><td>&amp;mdash;</td><td>Non-transgenic control line used to validate specificity of nitroreductase-dependent effects</td></tr><tr><td>Danio rerio (Tg(th:Gal4; UAS:NTR-mCherry))</td><td>&amp;mdash;</td><td>&amp;mdash;</td><td>Transgenic zebrafish line expressing nitroreductase-mCherry in dopaminergic neurons for targeted chemogenetic ablation</td></tr><tr><td>Dimethyl sulfoxide (DMSO), anhydrous (&amp;ge;99.9%)</td><td>Sigma-Aldrich</td><td>472301-500ML</td><td>Solvent for PTU, MTZ, and screening compounds</td></tr><tr><td>EthoVision XT</td><td>Noldus</td><td>RRID: SCR_000441</td><td>Behavioral tracking software used to quantify larval locomotion</td></tr><tr><td>Fiji (ImageJ)</td><td>NIH</td><td>RRID: SCR_003070</td><td>Image analysis software used for fluorescence quantification</td></tr><tr><td>Fine-tipped forceps or 0.25 mm probe</td><td>World Precision Instruments</td><td>WPI0118</td><td>Used to orient larvae during agarose embedding</td></tr><tr><td>Fluorescence microscope</td><td>Zeiss</td><td>11845460</td><td>Used to image dopaminergic neurons via mCherry fluorescence</td></tr><tr><td>Gemma Micro 150 ZF (juvenile diet, 100&amp;ndash;200 &amp;mu;m)</td><td>Skretting</td><td>10818945</td><td>Used to feed juvenile zebrafish</td></tr><tr><td>Gemma Micro 75 ZF (larval diet, 50&amp;ndash;100 &amp;mu;m)</td><td>Skretting</td><td>10818935</td><td>Used to feed larval zebrafish</td></tr><tr><td>GraphPad Prism</td><td>GraphPad</td><td>RRID: SCR_002798</td><td>Statistical analysis software used for data analysis and plotting</td></tr><tr><td>Heat block (42&amp;deg;C and 70&amp;deg;C capable)</td><td>Thermo Fisher</td><td>88870001</td><td>Used to melt and maintain agarose at controlled temperatures</td></tr><tr><td>High-resolution camera (top-down mount compatible)</td><td>Basler</td><td>88-327</td><td>Used to record larval locomotor activity</td></tr><tr><td>Hydrochloric acid (HCl), 0.1 M</td><td>Fisher Scientific</td><td>SA48-500</td><td>Used to adjust pH during tricaine preparation</td></tr><tr><td>Incubator (28.5&amp;deg;C, 14 h light:10 h dark photoperiod)</td><td>Millipore</td><td>Z763314</td><td>Maintains controlled environmental conditions for zebrafish development</td></tr><tr><td>Instant Ocean sea salt</td><td>Instant Ocean</td><td>SS15-10</td><td>Used to prepare blue egg water (BEW)</td></tr><tr><td>Light diffuser sheet (or equivalent)</td><td>Edmund Optics</td><td>27-103</td><td>Used to provide uniform lighting and minimize glare during behavioral recording</td></tr><tr><td>Light pad</td><td>Lightcraft</td><td>LC2004LED</td><td>Provides bottom illumination during locomotor tracking</td></tr><tr><td>Metronidazole (MTZ)</td><td>Sigma-Aldrich</td><td>M3761-100G</td><td>Used for chemogenetic ablation of dopaminergic neurons</td></tr><tr><td>Methylene blue, 1% aqueous solution</td><td>Sigma-Aldrich</td><td>M9140</td><td>Added to BEW to prevent microbial growth in embryos</td></tr><tr><td>Microcentrifuge tubes, 1.5 mL</td><td>Eppendorf</td><td>22363204</td><td>Used for reagent aliquoting and storage</td></tr><tr><td>Micropipette, P200</td><td>Gilson</td><td>FP10005S</td><td>Used for handling larvae and preparing solutions</td></tr><tr><td>Petri dishes, 100 mm</td><td>Corning</td><td>CLS430167</td><td>Used for embryo collection and incubation</td></tr><tr><td>Pipette tips, 200 &amp;mu;L</td><td>RAININ</td><td>30389188</td><td>Used with micropipettes for liquid handling and larval transfer</td></tr><tr><td>Sodium chloride (NaCl), &amp;ge;99.0%</td><td>Sigma-Aldrich</td><td>S9888-500G</td><td>Used for preparing buffer and aqueous solutions</td></tr><tr><td>Sodium hydroxide (NaOH), 0.1 M</td><td>Fisher Scientific</td><td>SS256-500</td><td>Used to adjust pH during tricaine preparation</td></tr><tr><td>Stereomicroscope</td><td>Nikon</td><td>SMZ-745</td><td>Used for larval manipulation and orientation during embedding</td></tr><tr><td>Temperature-controlled heat mat (25&amp;deg;C&amp;ndash;28&amp;deg;C)</td><td>Vivosun</td><td>V-SHMDT</td><td>Maintains stable temperature during behavioral recording</td></tr><tr><td>Tricaine methanesulfonate (MS-222)</td><td>Sigma-Aldrich</td><td>E10521-10G</td><td>Used as an anesthetic for zebrafish larvae</td></tr><tr><td>Tris base</td><td>Fisher Scientific</td><td>BP152-1</td><td>Used in preparation of buffered tricaine solution</td></tr><tr><td>Water purification system</td><td>Millipore</td><td>ZIQ7003T0</td><td>Used to generate deionized water for solution preparation</td></tr><tr><td>Zebrafish holding tanks</td><td>Aquaneering</td><td>ZS560</td><td>Used for maintaining adult zebrafish populations</td></tr><tr><td>1-Phenyl-2-thiourea (PTU)</td><td>Sigma-Aldrich</td><td>P7629-10G</td><td>Used to inhibit melanogenesis and maintain optical transparency</td></tr><tr><td>6-well plate, flat bottom</td><td>Corning</td><td>07-200-83</td><td>Used for locomotor behavioral assays</td></tr><tr><td>96-well plate, flat bottom</td><td>Greiner Bio-One</td><td>655096</td><td>Used for high-throughput imaging and screening assays</td></tr></tbody></table>

in vivo phenotyping of dopaminergic neurodegeneration in zebrafish larvae using behavioral analysis and high-content imaging

In Vivo Phenotyping of Dopaminergic Neurodegeneration in Zebrafish Larvae Using Behavioral Analysis and High-Content Imaging

Drug discovery research in neurodegeneration is constrained by the high cost and low throughput of traditional mammalian models. This bottleneck is particularly observed in Parkinson&#8217;s disease research, where rigorous and scalable dopaminergic (DA) neurodegeneration studies remain slow and resource intensive. To address this gap, we present a standardized, high-throughput phenotyping pipeline using a transgenic zebrafish model expressing nitroreductase in DA neurons to study DA neuron loss within five days post-fertilization. Zebrafish offer key advantages for translational neuroscience, including rapid larval development, optical transparency that enables in vivo whole-brain imaging, strong conservation of Parkinson&#8217;s disease-relevant genes and pathways, and intact neural circuitry not accessible in cell culture models. Our protocol integrates chemogenetic ablation and high-content imaging to generate rapid datasets for screening. DA neurons are selectively ablated using metronidazole (MTZ), producing specific and tunable neurodegeneration. MTZ treatment produces dose-dependent reductions in locomotion consistent with bradykinesia-like phenotypes, providing a robust behavioral correlate to DA cell loss. Since Parkinson&#8217;s disease is fundamentally a motor disorder, we pair anatomical measurements with functional behavioral readouts. Locomotor activity is recorded directly in a plate and quantified using automated tracking, extracting metrics including total distance traveled, swim bout frequency, and burst initiation. Zebrafish provide an efficient and scalable model system in which hundreds of larvae can be assayed simultaneously with minimal handling. We optimized a high-throughput, plate-based drug screening protocol designed to minimize experimental variance in undergraduate research. This standardized, plate-based format for screening candidate compounds for neuroprotection or functional rescue ensures that data collected by different researchers remains statistically comparable and effective for identifying neuroprotective candidates, improving reproducibility. Together, this methodology provides a robust, scalable framework for neurodegeneration research.

MTZ Treatment Produces Dose-Dependent Dopaminergic Neuron Loss and Locomotor Impairment
To determine the optimal parameters for MTZ-mediated ablation of DA neurons, fluorescence intensity, used as a proxy for DA neuron survival, and locomotor distance were evaluated across MTZ concentrations of 3, 4.5, and 9 mM following 24 h treatment. Each treatment group consisted of n = 10 individual Tg(th:Gal4; UAS:NTR-mCherry) larvae with a matched vehicle control group (1% DMSO in BEW w...

This protocol provides a standardized, high-throughput compound screening method using in vivo zebrafish to quantify dopaminergic neuron loss and locomotor behavioral deficits, enabling efficient screening of neuroprotective compounds.

Drug discovery research in neurodegeneration is constrained by the high cost and low throughput of traditional mammalian models. This bottleneck is ...

in-vivo-phenotyping-dopaminergic-neurodegeneration-zebrafish-larvae

Research

JoVE Journal

Neuroscience

A protocol to cut neurons in C. elegans with a MicroPoint  pulsed laser is presented. We describe setting up the system, immobilizing worms, and severing labeled neurons. Advantages include a relatively low-cost system and the ability to sever neuronal processes or ablate cells in vivo.

In vivo Laser Axotomy in C. elegans

A technique is described for broadly opening the blood-brain barrier in the mouse using microbubbles and ultrasound. Using this technique, manganese can be administered to the mouse brain. Because manganese is an MRI contrast agent that accumulates in depolarized neurons, this approach enables imaging of neuronal activity.

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain.  As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

Our laboratory developed a novel high-throughput automated imaging system that is useful for the detection of several different behaviors in 7-day-old zebrafish larvae. The system can be used for detecting subtle changes in behavior after the larvae have been exposed to environmental toxicants or pharmaceuticals.

Automated High-throughput Behavioral Analyses in Zebrafish Larvae

We present a method for the electroretinographic (ERG) analysis of zebrafish larvae utilizing micromanipulation and electroretinography techniques. This is a simple and straightforward method for assaying visual function of zebrafish larvae in vivo.

Electroretinogram Analysis of the Visual Response in Zebrafish Larvae

Here, we present a protocol to ablate a genetically labeled subpopulation of neurons by a two-photon laser from Zebrafish larvae.

Ablation of a Neuronal Population Using a Two-photon Laser and Its Assessment Using Calcium Imaging and Behavioral Recording in Zebrafish Larvae

We have developed a simple and adaptable workflow to extract quantitative data from fluorescence-imaging-based cell biological studies of protein aggregation and autophagic flux in the central nervous system of Drosophila models of neurodegeneration.

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ

Here, we present a protocol for the development and the characterization of a zebrafish model of epilepsy resulting from the transient inhibition of the DEPDC5 gene.

Behavioral And Physiological Analysis In A Zebrafish Model Of Epilepsy

Here we present an automated method for semi-quantitative determination of dopaminergic neuron number in the rat substantia nigra pars compacta.

Semi-Quantitative Determination of Dopaminergic Neuron Density in the Substantia Nigra of Rodent Models using Automated Image Analysis

Here we present a method to image the zebrafish embryonic brain in vivo upto larval and juvenile stages. This microinvasive procedure, adapted from electrophysiological approaches, provides access to cellular and subcellular details of mature neuron and can be combined with optogenetics and neuropharmacological studies for characterizing brain function and drug intervention.

In vivo Imaging of Fully Active Brain Tissue in Awake Zebrafish Larvae and Juveniles by Skull and Skin Removal

Discover a high-throughput phenotyping pipeline using transgenic zebrafish to study dopaminergic neuron loss in Parkinson's disease. This method integrates chemogenetic ablation with behavioral analysis and high-content imaging for rapid drug screening.

In Vivo Phenotyping of Dopaminergic Neurodegeneration in Zebrafish Larvae Using Behavioral Analysis and High-Content Imaging

Explore More Videos