Photoactivatable Genetically-Encoded Calcium Indicators for targeted neuronal imaging

Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca2+ indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca2+ detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).

[1]  Takeharu Nagai,et al.  Highlightable Ca2+ indicators for live cell imaging. , 2013, Journal of the American Chemical Society.

[2]  Jun Noguchi,et al.  Spine-Neck Geometry Determines NMDA Receptor-Dependent Ca2+ Signaling in Dendrites , 2005, Neuron.

[3]  Jyothi Arikkath,et al.  Culturing pyramidal neurons from the early postnatal mouse hippocampus and cortex , 2012, Nature Protocols.

[4]  Diego A. Pacheco,et al.  Fast GCaMPs for improved tracking of neuronal activity , 2013, Nature Communications.

[5]  M. Whitaker.  Genetically encoded probes for measurement of intracellular calcium. , 2010, Methods in cell biology.

[6]  E. Isacoff,et al.  Two-photon brightness of azobenzene photoswitches designed for glutamate receptor optogenetics , 2015, Proceedings of the National Academy of Sciences.

[7]  Roberto Araya,et al.  The spine neck filters membrane potentials , 2006, Proceedings of the National Academy of Sciences.

[8]  Massimo Scanziani,et al.  In vivo Labeling of Constellations of Functionally Identified Neurons for Targeted in vitro Recordings , 2011, Front. Neural Circuits.

[9]  Takeharu Nagai,et al.  Improved orange and red Ca²± indicators and photophysical considerations for optogenetic applications. , 2013, ACS chemical neuroscience.

[10]  Richard D. Fetter,et al.  wishful thinking Encodes a BMP Type II Receptor that Regulates Synaptic Growth in Drosophila , 2002, Neuron.

[11]  Jessica A. Cardin,et al.  Noninvasive optical inhibition with a red-shifted microbial rhodopsin , 2014, Nature Neuroscience.

[12]  Kristin Scott,et al.  Imaging Taste Responses in the Fly Brain Reveals a Functional Map of Taste Category and Behavior , 2006, Neuron.

[13]  Alexander Borst,et al.  A FRET-based calcium biosensor with fast signal kinetics and high fluorescence change. , 2006, Biophysical journal.

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

[15]  R. Zucker.  Calcium- and activity-dependent synaptic plasticity , 1999, Current Opinion in Neurobiology.

[16]  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.

[17]  Amy E Palmer,et al.  Measuring calcium dynamics in living cells with genetically encodable calcium indicators. , 2008, Methods.

[18]  Junichi Nakai,et al.  Real-Time Visualization of Neuronal Activity during Perception , 2013, Current Biology.

[19]  T. Hughes,et al.  Simultaneous Detection of Ca2+ and Diacylglycerol Signaling in Living Cells , 2012, PloS one.

[20]  T. Hara,et al.  Structure of the human-heart fatty-acid-binding protein 3 in complex with the fluorescent probe 1-anilinonaphthalene-8-sulphonic acid , 2013, Journal of synchrotron radiation.

[21]  D. Kleinfeld,et al.  ReaChR: A red-shifted variant of channelrhodopsin enables deep transcranial optogenetic excitation , 2013, Nature Neuroscience.

[22]  Kristin Scott,et al.  Taste Representations in the Drosophila Brain , 2004, Cell.

[23]  Pierre Drapeau,et al.  Motoneuron Activity Patterns Related to the Earliest Behavior of the Zebrafish Embryo , 2000, The Journal of Neuroscience.

[24]  R. W. Draft,et al.  Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system , 2007, Nature.

[25]  M. Ohkura,et al.  A high signal-to-noise Ca2+ probe composed of a single green fluorescent protein , 2001, Nature Biotechnology.

[26]  Akira Fushiki,et al.  Development of larval motor circuits in Drosophila , 2012, Development, growth & differentiation.

[27]  Kazuki Tainaka,et al.  A single circularly permuted GFP sensor for inositol-1,3,4,5-tetrakisphosphate based on a split PH domain. , 2009, Bioorganic & medicinal chemistry.

[28]  Konstantin A Lukyanov,et al.  A genetically encoded photosensitizer , 2006, Nature Biotechnology.

[29]  George H. Patterson,et al.  A Photoactivatable GFP for Selective Photolabeling of Proteins and Cells , 2002, Science.

[30]  S. Rumpel,et al.  Transgenic Mouse Models Enabling Photolabeling of Individual Neurons In Vivo , 2013, PloS one.

[31]  Takeharu Nagai,et al.  Highlightable Ca 2 + Indicators for Live Cell Imaging , 2013 .

[32]  T. Branco,et al.  Local Dendritic Activity Sets Release Probability at Hippocampal Synapses , 2008, Neuron.

[33]  R. Tsien,et al.  Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin , 1997, Nature.

[34]  Yongxin Zhao,et al.  An Expanded Palette of Genetically Encoded Ca2+ Indicators , 2011, Science.

[35]  Lin Tian,et al.  Genetically encoded fluorescent sensors for studying healthy and diseased nervous systems. , 2008, Drug discovery today. Disease models.

[36]  M. Drobizhev,et al.  Two-photon absorption properties of fluorescent proteins , 2011, Nature Methods.

[37]  Baljit S Khakh,et al.  A genetically targeted optical sensor to monitor calcium signals in astrocyte processes , 2010, Nature Neuroscience.

[38]  Ian R. Wickersham,et al.  Retrograde neuronal tracing with a deletion-mutant rabies virus , 2007, Nature Methods.

[39]  J. Brusés,et al.  In Vivo labeling of zebrafish motor neurons using an mnx1 enhancer and Gal4/UAS , 2011, Genesis.

[40]  Christian Griesinger,et al.  Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes , 2014, Nature Methods.

[41]  Yuji Ikegaya,et al.  Genetically Encoded Green Fluorescent Ca2+ Indicators with Improved Detectability for Neuronal Ca2+ Signals , 2012, PloS one.

[42]  Herwig Baier,et al.  Remote Control of Neuronal Activity with a Light-Gated Glutamate Receptor , 2007, Neuron.

[43]  K. Deisseroth,et al.  CLARITY for mapping the nervous system , 2013, Nature Methods.

[44]  Mark T. Harnett,et al.  An optimized fluorescent probe for visualizing glutamate neurotransmission , 2013, Nature Methods.

[45]  Stefan R. Pulver,et al.  Independent Optical Excitation of Distinct Neural Populations , 2014, Nature Methods.

[46]  Guangju Ji,et al.  Structural insight into enhanced calcium indicator GCaMP3 and GCaMPJ to promote further improvement , 2013, Protein & Cell.

[47]  K. Mikoshiba,et al.  Quantitative comparison of novel GCaMP-type genetically encoded Ca2+ indicators in mammalian neurons , 2012, Front. Cell. Neurosci..

[48]  Kevin J Mann,et al.  Dopaminergic Modulation of Sucrose Acceptance Behavior in Drosophila , 2012, Neuron.

[49]  S. R. Cajal,et al.  Estructura de los centros nerviosos de las Aves , 1888 .

[50]  Allan R. Jones,et al.  A mesoscale connectome of the mouse brain , 2014, Nature.

[51]  Michael Z. Lin,et al.  High-fidelity optical reporting of neuronal electrical activity with an ultrafast fluorescent voltage sensor , 2014, Nature Neuroscience.

[52]  E. Papagiakoumou,et al.  Two-photon optogenetics. , 2012, Progress in brain research.

[53]  E. Isacoff,et al.  Genetically encoded fluorescent sensors of membrane potential , 2008, Brain cell biology.

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

[55]  Imaging functional neural circuits in zebrafish with a new GCaMP and the Gal4FF-UAS system. , 2011, Communicative & integrative biology.

[56]  Alberto Diaspro,et al.  Two-photon activation and excitation properties of PA-GFP in the 720-920-nm region. , 2005, Biophysical journal.

[57]  Rafael Yuste,et al.  Nanotools for neuroscience and brain activity mapping. , 2013, ACS nano.

[58]  Hiroshi Kikuta,et al.  Transgenesis in zebrafish with the tol2 transposon system. , 2009, Methods in molecular biology.

[59]  T. Matsuda,et al.  Highlighted Ca2+ imaging with a genetically encoded ‘caged’ indicator , 2013, Scientific Reports.

[60]  Suliana Manley,et al.  Photoactivatable mCherry for high-resolution two-color fluorescence microscopy , 2009, Nature Methods.

[61]  L. Tian,et al.  Imaging neuronal activity with genetically encoded calcium indicators. , 2012, Cold Spring Harbor protocols.

[62]  S. Remington,et al.  Structure and mechanism of the photoactivatable green fluorescent protein. , 2009, Journal of the American Chemical Society.

[63]  M. Berridge,et al.  The versatility and universality of calcium signalling , 2000, Nature Reviews Molecular Cell Biology.

[64]  W. Gehring,et al.  Genetic control of development of the mushroom bodies, the associative learning centers in the Drosophila brain, by the eyeless, twin of eyeless, and Dachshund genes. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[65]  Aleksander Rebane,et al.  A long Stokes shift red fluorescent Ca2+ indicator protein for two-photon and ratiometric imaging , 2014, Nature Communications.

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

[67]  Richard Axel,et al.  A dimorphic pheromone circuit in Drosophila from sensory input to descending output , 2010, Nature.

[68]  J. Lippincott-Schwartz,et al.  Bright monomeric photoactivatable red fluorescent protein for two-color super-resolution sptPALM of live cells. , 2010, Journal of the American Chemical Society.

[69]  J. Livet,et al.  A technicolour approach to the connectome , 2008, Nature Reviews Neuroscience.

[70]  E. S. Ruthazer,et al.  Bulk electroporation of retinal ganglion cells in live Xenopus tadpoles. , 2013, Cold Spring Harbor protocols.