Whole-central nervous system functional imaging in larval Drosophila

Understanding how the brain works in tight concert with the rest of the central nervous system (CNS) hinges upon knowledge of coordinated activity patterns across the whole CNS. We present a method for measuring activity in an entire, non-transparent CNS with high spatiotemporal resolution. We combine a light-sheet microscope capable of simultaneous multi-view imaging at volumetric speeds 25-fold faster than the state-of-the-art, a whole-CNS imaging assay for the isolated Drosophila larval CNS and a computational framework for analysing multi-view, whole-CNS calcium imaging data. We image both brain and ventral nerve cord, covering the entire CNS at 2 or 5 Hz with two- or one-photon excitation, respectively. By mapping network activity during fictive behaviours and quantitatively comparing high-resolution whole-CNS activity maps across individuals, we predict functional connections between CNS regions and reveal neurons in the brain that identify type and temporal state of motor programs executed in the ventral nerve cord.

[1]  H. Atwood,et al.  Modular neuropile organization in the Drosophila larval brain facilitates identification and mapping of central neurons , 2006, The Journal of comparative neurology.

[2]  Sreekanth H. Chalasani,et al.  Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators , 2009, Nature Methods.

[3]  Benjamin H. White,et al.  Focusing Transgene Expression in Drosophila by Coupling Gal4 With a Novel Split-LexA Expression System , 2011, Genetics.

[4]  Philipp J. Keller,et al.  Fast, high-contrast imaging of animal development with scanned light sheet–based structured-illumination microscopy , 2010, Nature Methods.

[5]  Brian B. Avants,et al.  Explicit B-spline regularization in diffeomorphic image registration , 2013, Front. Neuroinform..

[6]  Misha B. Ahrens,et al.  Visualizing Whole-Brain Activity and Development at the Single-Cell Level Using Light-Sheet Microscopy , 2015, Neuron.

[7]  Kristin Branson,et al.  A multilevel multimodal circuit enhances action selection in Drosophila , 2015, Nature.

[8]  W. Kristan,et al.  Cellular substrates of action selection: a cluster of higher-order descending neurons shapes body posture and locomotion , 2008, Journal of Comparative Physiology A.

[9]  Justin Senseney,et al.  Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy , 2013, Nature Biotechnology.

[10]  K. Kawakami,et al.  Targeted gene expression by the Gal4‐UAS system in zebrafish , 2008, Development, Growth and Differentiation.

[11]  Matthias Landgraf,et al.  Charting the Drosophila neuropile: a strategy for the standardised characterisation of genetically amenable neurites. , 2003, Developmental biology.

[12]  V. Pieribone,et al.  Genetically Targeted Optical Electrophysiology in Intact Neural Circuits , 2013, Cell.

[13]  Germán Sumbre,et al.  Fast functional imaging of multiple brain regions in intact zebrafish larvae using Selective Plane Illumination Microscopy , 2013, BMC Neuroscience.

[14]  Julie H. Simpson,et al.  A GAL4-driver line resource for Drosophila neurobiology. , 2012, Cell reports.

[15]  Philipp J. Keller,et al.  Reconstruction of Zebrafish Early Embryonic Development by Scanned Light Sheet Microscopy , 2008, Science.

[16]  R. Yuste,et al.  The Brain Activity Map Project and the Challenge of Functional Connectomics , 2012, Neuron.

[17]  T. Holy,et al.  Image-based calibration of a deformable mirror in wide-field microscopy. , 2010, Applied optics.

[18]  Philipp J. Keller,et al.  Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy , 2012, Nature Methods.

[19]  M. Suster,et al.  Embryonic assembly of a central pattern generator without sensory input , 2002, Nature.

[20]  R. Ritzmann,et al.  Descending control of turning behavior in the cockroach, Blaberus discoidalis , 2007, Journal of Comparative Physiology A.

[21]  E. Boyden,et al.  Simultaneous whole-animal 3D-imaging of neuronal activity using light-field microscopy , 2014, Nature Methods.

[22]  Sen-Lin Lai,et al.  Genetic mosaic with dual binary transcriptional systems in Drosophila , 2006, Nature Neuroscience.

[23]  Philipp J. Keller,et al.  Whole-brain functional imaging at cellular resolution using light-sheet microscopy , 2013, Nature Methods.

[24]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[25]  D. Soll,et al.  Coordination and Modulation of Locomotion Pattern Generators in Drosophila Larvae: Effects of Altered Biogenic Amine Levels by the Tyramine β Hydroxlyase Mutation , 2006, The Journal of Neuroscience.

[26]  Diwakar Turaga,et al.  Functional organization of glomerular maps in the mouse accessory olfactory bulb , 2014, Nature Neuroscience.

[27]  Byron M. Yu,et al.  Dimensionality reduction for large-scale neural recordings , 2014, Nature Neuroscience.

[28]  Julie H. Simpson,et al.  Genetic Manipulation of Genes and Cells in the Nervous System of the Fruit Fly , 2011, Neuron.

[29]  John B. Thomas,et al.  A sensory feedback circuit coordinates muscle activity in Drosophila , 2007, Molecular and Cellular Neuroscience.

[30]  T. Murphy,et al.  Motor maps and the cortical control of movement , 2014, Current Opinion in Neurobiology.

[31]  Drew N. Robson,et al.  Brain-wide neuronal dynamics during motor adaptation in zebrafish , 2012, Nature.

[32]  Anthony Santella,et al.  A hybrid blob-slice model for accurate and efficient detection of fluorescence labeled nuclei in 3D , 2010, BMC Bioinformatics.

[33]  Omotara Ogundeyi,et al.  A GAL4 driver resource for developmental and behavioral studies on the larval CNS of Drosophila. , 2014, Cell reports.

[34]  Todd Anderson,et al.  Elastic source selection for in vivo imaging of neuronal ensembles , 2009, 2009 IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[35]  Liang Liang,et al.  The Q System: A Repressible Binary System for Transgene Expression, Lineage Tracing, and Mosaic Analysis , 2010, Cell.

[36]  Marta Zlatic,et al.  Positional Cues in the Drosophila Nerve Cord: Semaphorins Pattern the Dorso-Ventral Axis , 2009, PLoS biology.

[37]  H. Bellen,et al.  Genome-wide manipulations of Drosophila melanogaster with transposons, Flp recombinase, and ΦC31 integrase. , 2012, Methods in molecular biology.

[38]  T. Holy,et al.  Fast Three-Dimensional Fluorescence Imaging of Activity in Neural Populations by Objective-Coupled Planar Illumination Microscopy , 2008, Neuron.

[39]  Kevin L. Briggman,et al.  Optical Imaging of Neuronal Populations During Decision-Making , 2005, Science.

[40]  Kevin W Eliceiri,et al.  NIH Image to ImageJ: 25 years of image analysis , 2012, Nature Methods.

[41]  Takashi Kawashima,et al.  Mapping brain activity at scale with cluster computing , 2014, Nature Methods.

[42]  Diwakar Turaga,et al.  Aberrations and their correction in light-sheet microscopy: a low-dimensional parametrization. , 2013, Biomedical optics express.

[43]  James W. Truman,et al.  Transvection Is Common Throughout the Drosophila Genome , 2012, Genetics.

[44]  A. J. Pollack,et al.  Neural Activity in the Central Complex of the Insect Brain Is Linked to Locomotor Changes , 2010, Current Biology.

[45]  K. Okkenhaug,et al.  Gene targeting in mice: a review. , 2013, Methods in molecular biology.

[46]  Brian B. Avants,et al.  The optimal template effect in hippocampus studies of diseased populations , 2010, NeuroImage.

[47]  M. Orger,et al.  Whole-Brain Activity Maps Reveal Stereotyped, Distributed Networks for Visuomotor Behavior , 2014, Neuron.

[48]  I. Ial,et al.  Nature Communications , 2010, Nature Cell Biology.

[49]  David S. Koos,et al.  Deep and fast live imaging with two-photon scanned light-sheet microscopy , 2011, Nature Methods.

[50]  R. Mann,et al.  Swept confocally-aligned planar excitation (SCAPE) microscopy for high speed volumetric imaging of behaving organisms , 2014, Nature Photonics.

[51]  Citlali Pérez Campos,et al.  High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics , 2013, Nature Communications.

[52]  Stefan R. Pulver,et al.  Ultra-sensitive fluorescent proteins for imaging neuronal activity , 2013, Nature.

[53]  Lars Hufnagel,et al.  Multiview light-sheet microscope for rapid in toto imaging , 2012, Nature Methods.

[54]  F. Del Bene,et al.  Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy , 2004, Science.

[55]  Philipp J. Keller,et al.  Light-sheet functional imaging in fictively behaving zebrafish , 2014, Nature Methods.