Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter

We combine a chemically-synthesized, voltage-sensitive fluorophore with a genetically encoded, self-labeling enzyme to enable voltage imaging in Drosophila melanogaster. Previously, we showed that a rhodamine voltage reporter (RhoVR) combined with the HaloTag self-labeling enzyme could be used to mo...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Molly J. Kirk, Brittany R. Benlian, Yifu Han, Arya Gold, Ashvin Ravi, Parker E. Deal, Rosana S. Molina, Mikhail Drobizhev, Dion Dickman, Kristin Scott, Evan W. Miller
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	imaging fluorescence voltage Drosophila neuromuscular junction (NMJ) Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
Acceso en línea:	https://doaj.org/article/8bb0c427fcf548cfaf73e51d8ca6a5f4
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:8bb0c427fcf548cfaf73e51d8ca6a5f4
record_format	dspace
spelling	oai:doaj.org-article:8bb0c427fcf548cfaf73e51d8ca6a5f42021-11-16T07:46:49ZVoltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter1662-453X10.3389/fnins.2021.754027https://doaj.org/article/8bb0c427fcf548cfaf73e51d8ca6a5f42021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fnins.2021.754027/fullhttps://doaj.org/toc/1662-453XWe combine a chemically-synthesized, voltage-sensitive fluorophore with a genetically encoded, self-labeling enzyme to enable voltage imaging in Drosophila melanogaster. Previously, we showed that a rhodamine voltage reporter (RhoVR) combined with the HaloTag self-labeling enzyme could be used to monitor membrane potential changes from mammalian neurons in culture and brain slice. Here, we apply this hybrid RhoVR-Halo approach in vivo to achieve selective neuron labeling in intact fly brains. We generate a Drosophila UAS-HaloTag reporter line in which the HaloTag enzyme is expressed on the surface of cells. We validate the voltage sensitivity of this new construct in cell culture before driving expression of HaloTag in specific brain neurons in flies. We show that selective labeling of synapses, cells, and brain regions can be achieved with RhoVR-Halo in either larval neuromuscular junction (NMJ) or in whole adult brains. Finally, we validate the voltage sensitivity of RhoVR-Halo in fly tissue via dual-electrode/imaging at the NMJ, show the efficacy of this approach for measuring synaptic excitatory post-synaptic potentials (EPSPs) in muscle cells, and perform voltage imaging of carbachol-evoked depolarization and osmolarity-evoked hyperpolarization in projection neurons and in interoceptive subesophageal zone neurons in fly brain explants following in vivo labeling. We envision the turn-on response to depolarizations, fast response kinetics, and two-photon compatibility of chemical indicators, coupled with the cellular and synaptic specificity of genetically-encoded enzymes, will make RhoVR-Halo a powerful complement to neurobiological imaging in Drosophila.Molly J. KirkBrittany R. BenlianYifu HanArya GoldAshvin RaviParker E. DealRosana S. MolinaMikhail DrobizhevDion DickmanKristin ScottKristin ScottEvan W. MillerEvan W. MillerEvan W. MillerFrontiers Media S.A.articleimagingfluorescencevoltageDrosophilaneuromuscular junction (NMJ)Neurosciences. Biological psychiatry. NeuropsychiatryRC321-571ENFrontiers in Neuroscience, Vol 15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	imaging fluorescence voltage Drosophila neuromuscular junction (NMJ) Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
spellingShingle	imaging fluorescence voltage Drosophila neuromuscular junction (NMJ) Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Molly J. Kirk Brittany R. Benlian Yifu Han Arya Gold Ashvin Ravi Parker E. Deal Rosana S. Molina Mikhail Drobizhev Dion Dickman Kristin Scott Kristin Scott Evan W. Miller Evan W. Miller Evan W. Miller Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter
description	We combine a chemically-synthesized, voltage-sensitive fluorophore with a genetically encoded, self-labeling enzyme to enable voltage imaging in Drosophila melanogaster. Previously, we showed that a rhodamine voltage reporter (RhoVR) combined with the HaloTag self-labeling enzyme could be used to monitor membrane potential changes from mammalian neurons in culture and brain slice. Here, we apply this hybrid RhoVR-Halo approach in vivo to achieve selective neuron labeling in intact fly brains. We generate a Drosophila UAS-HaloTag reporter line in which the HaloTag enzyme is expressed on the surface of cells. We validate the voltage sensitivity of this new construct in cell culture before driving expression of HaloTag in specific brain neurons in flies. We show that selective labeling of synapses, cells, and brain regions can be achieved with RhoVR-Halo in either larval neuromuscular junction (NMJ) or in whole adult brains. Finally, we validate the voltage sensitivity of RhoVR-Halo in fly tissue via dual-electrode/imaging at the NMJ, show the efficacy of this approach for measuring synaptic excitatory post-synaptic potentials (EPSPs) in muscle cells, and perform voltage imaging of carbachol-evoked depolarization and osmolarity-evoked hyperpolarization in projection neurons and in interoceptive subesophageal zone neurons in fly brain explants following in vivo labeling. We envision the turn-on response to depolarizations, fast response kinetics, and two-photon compatibility of chemical indicators, coupled with the cellular and synaptic specificity of genetically-encoded enzymes, will make RhoVR-Halo a powerful complement to neurobiological imaging in Drosophila.
format	article
author	Molly J. Kirk Brittany R. Benlian Yifu Han Arya Gold Ashvin Ravi Parker E. Deal Rosana S. Molina Mikhail Drobizhev Dion Dickman Kristin Scott Kristin Scott Evan W. Miller Evan W. Miller Evan W. Miller
author_facet	Molly J. Kirk Brittany R. Benlian Yifu Han Arya Gold Ashvin Ravi Parker E. Deal Rosana S. Molina Mikhail Drobizhev Dion Dickman Kristin Scott Kristin Scott Evan W. Miller Evan W. Miller Evan W. Miller
author_sort	Molly J. Kirk
title	Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter
title_short	Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter
title_full	Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter
title_fullStr	Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter
title_full_unstemmed	Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter
title_sort	voltage imaging in drosophila using a hybrid chemical-genetic rhodamine voltage reporter
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/8bb0c427fcf548cfaf73e51d8ca6a5f4
work_keys_str_mv	AT mollyjkirk voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT brittanyrbenlian voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT yifuhan voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT aryagold voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT ashvinravi voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT parkeredeal voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT rosanasmolina voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT mikhaildrobizhev voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT diondickman voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT kristinscott voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT kristinscott voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT evanwmiller voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT evanwmiller voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter AT evanwmiller voltageimagingindrosophilausingahybridchemicalgeneticrhodaminevoltagereporter
_version_	1718426616449728512

Voltage Imaging in Drosophila Using a Hybrid Chemical-Genetic Rhodamine Voltage Reporter

Ejemplares similares