Ultrafast Two-Photon Imaging of a High-Gain Voltage Indicator in Awake Behaving Mice.
Cell
; 179(7): 1590-1608.e23, 2019 12 12.
Article
em En
| MEDLINE
| ID: mdl-31835034
ABSTRACT
Optical interrogation of voltage in deep brain locations with cellular resolution would be immensely useful for understanding how neuronal circuits process information. Here, we report ASAP3, a genetically encoded voltage indicator with 51% fluorescence modulation by physiological voltages, submillisecond activation kinetics, and full responsivity under two-photon excitation. We also introduce an ultrafast local volume excitation (ULoVE) method for kilohertz-rate two-photon sampling in vivo with increased stability and sensitivity. Combining a soma-targeted ASAP3 variant and ULoVE, we show single-trial tracking of spikes and subthreshold events for minutes in deep locations, with subcellular resolution and with repeated sampling over days. In the visual cortex, we use soma-targeted ASAP3 to illustrate cell-type-dependent subthreshold modulation by locomotion. Thus, ASAP3 and ULoVE enable high-speed optical recording of electrical activity in genetically defined neurons at deep locations during awake behavior.
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MEDLINE
Assunto principal:
Ritmo Teta
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Vigília
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Encéfalo
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Proteínas Ativadoras de GTPase
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Microscopia de Fluorescência por Excitação Multifotônica
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Optogenética
Limite:
Animals
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Female
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Humans
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Male
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article