Full-field exposure of larval zebrafish to narrow waveband LED light sources at defined power and energy for optogenetic applications.

Animals Zebrafish / physiology Optogenetics / methods Larva Neurons / physiology Transgenes
Energy FAP GAVPO Kaede LED MG2I Optogenetics Power Singlet oxygen Zebrafish dL5**

Journal

Journal of neuroscience methods
ISSN: 1872-678X
Titre abrégé: J Neurosci Methods
Pays: Netherlands
ID NLM: 7905558

Informations de publication

Date de publication:
01 Jan 2024
Historique:
received: 21 06 2023
revised: 15 10 2023
accepted: 27 10 2023
medline: 5 12 2023
pubmed: 2 11 2023
entrez: 1 11 2023
Statut: ppublish

Résumé

Optogenetic approaches in transparent zebrafish models have provided numerous insights into vertebrate neurobiology. The purpose of this study was to develop methods to activate light-sensitive transgene products simultaneously throughout an entire larval zebrafish. We developed a LED illumination stand and microcontroller unit to expose zebrafish larvae reproducibly to full field illumination at defined wavelength, power, and energy. The LED stand generated a sufficiently flat illumination field to expose multiple larval zebrafish to high power light stimuli uniformly, while avoiding sample bath warming. The controller unit allowed precise automated delivery of predetermined amounts of light energy at calibrated power. We demonstrated the utility of the approach by driving photoconversion of Kaede (398 nm), photodimerization of GAVPO (450 nm), and photoactivation of dL5**/MG2I (661 nm) in neurons throughout the CNS of larval zebrafish. Observed outcomes were influenced by both total light energy and its rate of delivery, highlighting the importance of controlling these variables to obtain reproducible results. Our approach employs inexpensive LED chip arrays to deliver narrow-waveband light with a sufficiently flat illumination field to span multiple larval zebrafish simultaneously. Calibration of light power and energy are built into the workflow. The LED illuminator and controller can be constructed from widely available materials using the drawings, instructions, and software provided. This approach will be useful for multiple optogenetic applications in zebrafish and other models.

Sections du résumé

BACKGROUND BACKGROUND
Optogenetic approaches in transparent zebrafish models have provided numerous insights into vertebrate neurobiology. The purpose of this study was to develop methods to activate light-sensitive transgene products simultaneously throughout an entire larval zebrafish.
NEW METHOD METHODS
We developed a LED illumination stand and microcontroller unit to expose zebrafish larvae reproducibly to full field illumination at defined wavelength, power, and energy.
RESULTS RESULTS
The LED stand generated a sufficiently flat illumination field to expose multiple larval zebrafish to high power light stimuli uniformly, while avoiding sample bath warming. The controller unit allowed precise automated delivery of predetermined amounts of light energy at calibrated power. We demonstrated the utility of the approach by driving photoconversion of Kaede (398 nm), photodimerization of GAVPO (450 nm), and photoactivation of dL5**/MG2I (661 nm) in neurons throughout the CNS of larval zebrafish. Observed outcomes were influenced by both total light energy and its rate of delivery, highlighting the importance of controlling these variables to obtain reproducible results.
COMPARISON WITH EXISTING METHODS METHODS
Our approach employs inexpensive LED chip arrays to deliver narrow-waveband light with a sufficiently flat illumination field to span multiple larval zebrafish simultaneously. Calibration of light power and energy are built into the workflow.
CONCLUSIONS CONCLUSIONS
The LED illuminator and controller can be constructed from widely available materials using the drawings, instructions, and software provided. This approach will be useful for multiple optogenetic applications in zebrafish and other models.

Identifiants

DOI: 10.1016/j.jneumeth.2023.110001 PMID: 37914002
pubmed: 37914002
pii: S0165-0270(23)00220-0
doi: 10.1016/j.jneumeth.2023.110001
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

110001

Subventions

Organisme : NINDS NIH HHS
ID : R33 NS125280
Pays : United States
Organisme : NINDS NIH HHS
ID : R61 NS125280
Pays : United States

Informations de copyright

Published by Elsevier B.V.

Déclaration de conflit d'intérêts

Declaration of Competing Interest The authors have no conflicts of interest to declare. AI was not used at any stage in this research or in the preparation of this article.

Auteurs

Alexander H Burton (AH)

Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA; Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, PA, USA; Undergraduate Program in Chemical and Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA.

Binxuan Jiao (B)

Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA; Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, PA, USA; Tsinghua University Medical School, Beijing, China.

Qing Bai (Q)

Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA; Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, PA, USA.

Victor S Van Laar (VS)

Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA; Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, PA, USA.

Travis B Wheeler (TB)

Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA.

Simon C Watkins (SC)

Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA; Center for Biologic Imaging, University of Pittsburgh, Pittsburgh, PA, USA.

Marcel P Bruchez (MP)

Department of Chemistry, Carnegie Mellon University, Pittsburgh, PA, USA; Molecular Biosensors and Imaging Center, Carnegie Mellon University, Pittsburgh, PA, USA.

Edward A Burton (EA)

Department of Neurology, University of Pittsburgh, Pittsburgh, PA, USA; Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, PA, USA; Geriatric Research Education and Clinical Center, Pittsburgh VA Healthcare System, Pittsburgh, PA, USA. Electronic address: eab25@pitt.edu.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Full-field exposure of larval zebrafish to...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Poissons › Cypriniformes › Cyprinidae › Danio zébré › Full-field exposure of larval zebrafish to...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Optogénétique › Full-field exposure of larval zebrafish to...
questionsmedicales.fr › Phénomènes physiologiques › Croissance et développement › Morphogenèse › Métamorphose biologique › Étapes du cycle de vie › Larve › Full-field exposure of larval zebrafish to...
questionsmedicales.fr › Cellules › Neurones › Full-field exposure of larval zebrafish to...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Composants de génome › Gènes › Transgènes › Full-field exposure of larval zebrafish to...