A conditional GABAergic synaptic vesicle marker for Drosophila.

Animals Drosophila / metabolism GABAergic Neurons Glutamic Acid / metabolism Synapses / metabolism Synaptic Vesicles / metabolism
Drosophila Epitope tag GABAergic Synaptic vesicle VGAT

Journal

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

Informations de publication

Date de publication:
15 Apr 2022
Historique:
received: 31 12 2021
revised: 15 02 2022
accepted: 22 02 2022
pubmed: 28 2 2022
medline: 8 4 2022
entrez: 27 2 2022
Statut: ppublish

Résumé

Throughout the animal kingdom, GABA is the principal inhibitory neurotransmitter of the nervous system. It is essential for maintaining the homeostatic balance between excitation and inhibition required for the brain to operate normally. Identification of GABAergic neurons and their GABA release sites are thus essential for understanding how the brain regulates the excitability of neurons and the activity of neural circuits responsible for numerous aspects of brain function including information processing, locomotion, learning, memory, and synaptic plasticity, among others. Since the structure and features of GABA synapses are critical to understanding their function within specific neural circuits of interest, here we developed and characterized a conditional marker of GABAergic synaptic vesicles for Drosophila, 9XV5-vGAT. 9XV5-vGAT is validated for conditionality of expression, specificity for localization to synaptic vesicles, specificity for expression in GABAergic neurons, and functionality. Its utility for GABAergic neurotransmitter phenotyping and identification of GABA release sites was verified for ellipsoid body neurons of the central complex. In combination with previously reported conditional SV markers for acetylcholine and glutamate, 9XV5-vGAT was used to demonstrate fast neurotransmitter phenotyping of subesophageal ganglion neurons. This method is an alternative to single cell transcriptomics for neurotransmitter phenotyping and can be applied to any neurons of interest represented by a binary transcription system driver. A conditional GABAergic synaptic vesicle marker has been developed and validated for GABA neurotransmitter phenotyping and subcellular localization of GABAergic synaptic vesicles.

Sections du résumé

BACKGROUND BACKGROUND
Throughout the animal kingdom, GABA is the principal inhibitory neurotransmitter of the nervous system. It is essential for maintaining the homeostatic balance between excitation and inhibition required for the brain to operate normally. Identification of GABAergic neurons and their GABA release sites are thus essential for understanding how the brain regulates the excitability of neurons and the activity of neural circuits responsible for numerous aspects of brain function including information processing, locomotion, learning, memory, and synaptic plasticity, among others.
NEW METHOD METHODS
Since the structure and features of GABA synapses are critical to understanding their function within specific neural circuits of interest, here we developed and characterized a conditional marker of GABAergic synaptic vesicles for Drosophila, 9XV5-vGAT.
RESULTS RESULTS
9XV5-vGAT is validated for conditionality of expression, specificity for localization to synaptic vesicles, specificity for expression in GABAergic neurons, and functionality. Its utility for GABAergic neurotransmitter phenotyping and identification of GABA release sites was verified for ellipsoid body neurons of the central complex. In combination with previously reported conditional SV markers for acetylcholine and glutamate, 9XV5-vGAT was used to demonstrate fast neurotransmitter phenotyping of subesophageal ganglion neurons.
COMPARISON WITH EXISTING METHODS METHODS
This method is an alternative to single cell transcriptomics for neurotransmitter phenotyping and can be applied to any neurons of interest represented by a binary transcription system driver.
CONCLUSION CONCLUSIONS
A conditional GABAergic synaptic vesicle marker has been developed and validated for GABA neurotransmitter phenotyping and subcellular localization of GABAergic synaptic vesicles.

Identifiants

DOI: 10.1016/j.jneumeth.2022.109540 PMID: 35219770 PMC: PMC8940707
pubmed: 35219770
pii: S0165-0270(22)00067-X
doi: 10.1016/j.jneumeth.2022.109540
pmc: PMC8940707
mid: NIHMS1784480
pii:
doi:

Substances chimiques

Glutamic Acid 3KX376GY7L

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

109540

Subventions

Organisme : NIGMS NIH HHS
ID : R01 GM115510
Pays : United States

Informations de copyright

Copyright © 2022 Elsevier B.V. All rights reserved.

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Auteurs

Sarah J Certel (SJ)

Division of Biological Sciences, Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT, USA.

Brian D McCabe (BD)

Brain Mind Institute, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland.

R Steven Stowers (RS)

Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, USA. Electronic address: sstowers@montana.edu.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › A conditional GABAergic synaptic vesicle...
questionsmedicales.fr › Eucaryotes › Animaux › Invertébrés › Arthropodes › Insectes › Pterygota › Neoptera › Holometabola › Diptera › Drosophilidae › Drosophila › A conditional GABAergic synaptic vesicle...
questionsmedicales.fr › Cellules › Neurones › Neurones GABAergiques › A conditional GABAergic synaptic vesicle...
questionsmedicales.fr › Acides aminés, peptides et protéines › Acides aminés › Acides aminés excitateurs › Acide glutamique › A conditional GABAergic synaptic vesicle...
questionsmedicales.fr › Cellules › Structures cellulaires › Membrane cellulaire › Structures de la membrane cellulaire › Jonctions intercellulaires › Synapses › A conditional GABAergic synaptic vesicle...
questionsmedicales.fr › Cellules › Structures cellulaires › Espace intracellulaire › Cytoplasme › Structures cytoplasmiques › Organites › Vésicules cytoplasmiques › Vésicules de transport › Vésicules de sécrétion › Vésicules synaptiques › A conditional GABAergic synaptic vesicle...