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Figure 2. Whole-Brain Neuronal and Astrocyte Dynamics Show Ca2+ Increases in Specific Cell Populations during Futility-Induced Passivity For a Figure360 author presentation of this figure, see https://doi.org/10.1016/j.cell.2019.05.050. (A) Experimental setup. A light-sheet microscope recorded from most radial astrocytes and neurons in the brain at cellular resolution, using either single- population GCaMP6f or simultaneous dual-color GCaMP6f and jRGECO1b recordings, while fictive behavior was monitored and visual stimulus was delivered. (B) Whole-brain neuronal and glial activity before, at, and after passivity onset in an example <t>Tg(gfap:jRGECO1b);</t> Tg(elavl3:GCaMP6f) fish. Top: mean neuronal brain-wide activity was higher during active behavior than passivity. Bottom: conversely, mean brain-wide glial Ca2+ signals increased before passivity onset and remained elevated during passivity. A, anterior; P, posterior; D, dorsal; V, ventral. Images are maximum-intensity projections of means triggered on passivity onset (64 events). (C) Neuronal and glial signals near passivity onset, averaged across the brain of the same fish from (B). Average neuronal Ca2+ decayed after passivity onset, while average glial Ca2+ increased before passivity onset and peaked soon after.
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Figure 2. Whole-Brain Neuronal and Astrocyte Dynamics Show Ca2+ Increases in Specific Cell Populations during Futility-Induced Passivity For a Figure360 author presentation of this figure, see https://doi.org/10.1016/j.cell.2019.05.050. (A) Experimental setup. A light-sheet microscope recorded from most radial astrocytes and neurons in the brain at cellular resolution, using either single- population GCaMP6f or simultaneous dual-color GCaMP6f and jRGECO1b recordings, while fictive behavior was monitored and visual stimulus was delivered. (B) Whole-brain neuronal and glial activity before, at, and after passivity onset in an example Tg(gfap:jRGECO1b); Tg(elavl3:GCaMP6f) fish. Top: mean neuronal brain-wide activity was higher during active behavior than passivity. Bottom: conversely, mean brain-wide glial Ca2+ signals increased before passivity onset and remained elevated during passivity. A, anterior; P, posterior; D, dorsal; V, ventral. Images are maximum-intensity projections of means triggered on passivity onset (64 events). (C) Neuronal and glial signals near passivity onset, averaged across the brain of the same fish from (B). Average neuronal Ca2+ decayed after passivity onset, while average glial Ca2+ increased before passivity onset and peaked soon after.

Journal: Cell

Article Title: Glia Accumulate Evidence that Actions Are Futile and Suppress Unsuccessful Behavior.

doi: 10.1016/j.cell.2019.05.050

Figure Lengend Snippet: Figure 2. Whole-Brain Neuronal and Astrocyte Dynamics Show Ca2+ Increases in Specific Cell Populations during Futility-Induced Passivity For a Figure360 author presentation of this figure, see https://doi.org/10.1016/j.cell.2019.05.050. (A) Experimental setup. A light-sheet microscope recorded from most radial astrocytes and neurons in the brain at cellular resolution, using either single- population GCaMP6f or simultaneous dual-color GCaMP6f and jRGECO1b recordings, while fictive behavior was monitored and visual stimulus was delivered. (B) Whole-brain neuronal and glial activity before, at, and after passivity onset in an example Tg(gfap:jRGECO1b); Tg(elavl3:GCaMP6f) fish. Top: mean neuronal brain-wide activity was higher during active behavior than passivity. Bottom: conversely, mean brain-wide glial Ca2+ signals increased before passivity onset and remained elevated during passivity. A, anterior; P, posterior; D, dorsal; V, ventral. Images are maximum-intensity projections of means triggered on passivity onset (64 events). (C) Neuronal and glial signals near passivity onset, averaged across the brain of the same fish from (B). Average neuronal Ca2+ decayed after passivity onset, while average glial Ca2+ increased before passivity onset and peaked soon after.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-a2B- adrenergic receptor Life Technologies Cat# MA1-12721; RRID:AB_2225066 Bacterial and Virus Strains dbh-GFP-pDEST-ISce (Zhu et al., 2012b) N/A FCK-gene86-GFP (CoChR) (Klapoetke et al., 2014) N/A (Tol2-Isl-gal4vp16-4; UAS:TrpV1-tagRFpt) (Chen et al., 2016) N/A pEGFP-gfap (Intron1/50/Exon1-zebrafish) Addgene Cat# 39761 pcDNA3.1-opto-a1AR-EYFP Addgene Cat# 20947 Experimental Models: Organisms/Strains Tg(elavl3:GCaMP6f)jf1 Janelia Research Campus (JRC) (Freeman et al., 2014) N/A Tg(elavl3:jRGECO1b)jf17 JRC (Dana et al., 2016) N/A Tg(elavl3:H2B-GCaMP6f)jf7 JRC (Dunn et al., 2016) N/A Tg(gfap:jRGECO1b)jf33 JRC N/A Tg(gfap:GCaMP6f)jf22 JRC N/A Tg(gfap:CoChR-eGFP)jf29 JRC N/A Tg(gfap:opto-a1AR-eYFP)jf81 JRC N/A Tg(gfap:H2B-GCaMP6f)jf23 JRC N/A Tg(dbh:KalTA4)jf12 JRC N/A Tg(UAS:CoChR-eGFP)jf44 JRC N/A Tg(gfap:tdTomato)jf21 JRC N/A Tg(gad1b:Gal4)jf49 JRC N/A Tg(th-P2A-Gal4) Chinese Academy of Sciences (Li et al., 2015) N/A Tg(UAS:GCaMP6s) National Institute of Genetics N/A Tg(net:mTdTomato) Cornell University (Farrar et al., 2018) N/A Tg(net:H2B-GCaMP6f) Cornell University (Farrar et al., 2018) N/A Tg(gad1b:loxP-RFP-loxP-GFP) Okazaki Institute for Integrative Bioscience (Satou et al., 2013) N/A Software and Algorithms MATLAB MathWorks https://www.mathworks.com ImageJ National Institutes of Health https://imagej.nih.gov/ij/ C# (.NET Framework 3.5) (stimulus/behavior) Microsoft https://docs.microsoft.com/en-us/dotnet/ csharp/ Cell segmentation and component detection This study https://github.com/mikarubi/segmentation Chemicals, Peptides, and Recombinant Proteins Alpha-Bungarotoxin Sigma-Aldrich Cat# B137 Capsaicin Sigma-Aldrich Cat# M2028 Xestospongin C Sigma-Aldrich Cat# X2628 Prazosin hydrochloride Sigma-Aldrich Cat# P7791 Cyclazosin hydrochloride Sigma-Aldrich Cat# C247 Idazoxan hydrochloride Sigma-Aldrich Cat# I6138 Propranolol hydrochloride Sigma-Aldrich Cat# P0884 Methoxamine hydrochloride Sigma-Aldrich Cat# M6524 (Continued on next page) e1 Cell 178, 1–17.e1–e8, June 27, 2019

Techniques: Microscopy, Activity Assay