Probing the function of neuronal populations: Combining micromirror-based optogenetic photostimulation with voltage-sensitive dye imaging

Recent advances in our understanding of brain function have come from using light to either control or image neuronal activity. Here we describe an approach that combines both techniques: a micromirror array is used to photostimulate populations of presynaptic neurons expressing channelrhodopsin-2, while a red-shifted voltage-sensitive dye allows optical detection of resulting postsynaptic activity. Such technology allowed us to control the activity of cerebellar interneurons while simultaneously recording inhibitory responses in multiple Purkinje neurons, their postsynaptic targets. This approach should substantially accelerate our understanding of information processing by populations of neurons within brain circuits.

[1]  Raag D. Airan,et al.  Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures , 2010, Nature Protocols.

[2]  G. Feng,et al.  Cell type–specific channelrhodopsin-2 transgenic mice for optogenetic dissection of neural circuitry function , 2011, Nature Methods.

[3]  S. Scully,et al.  Evidence for a charge-shift electrochromic mechanism in a probe of membrane potential , 1979, Nature.

[4]  R. Traub,et al.  Synchronized oscillations in interneuron networks driven by metabotropic glutamate receptor activation , 1995, Nature.

[5]  E. Marder,et al.  Plasticity in single neuron and circuit computations , 2004, Nature.

[6]  Miles A Whittington,et al.  Cellular mechanisms of neuronal population oscillations in the hippocampus in vitro. , 2004, Annual review of neuroscience.

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

[8]  K. Deisseroth,et al.  Millisecond-timescale, genetically targeted optical control of neural activity , 2005, Nature Neuroscience.

[9]  Yosef Yarom,et al.  Stars and Stripes in the Cerebellar Cortex: A Voltage Sensitive Dye Study , 2007, Frontiers in systems neuroscience.

[10]  James M. Bower,et al.  Model-Founded Explorations of the Roles of Molecular Layer Inhibition in Regulating Purkinje Cell Responses in Cerebellar Cortex: More Trouble for the Beam Hypothesis , 2010, Front. Cell. Neurosci..

[11]  E. Bamberg,et al.  Channelrhodopsin-2, a directly light-gated cation-selective membrane channel , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Aravinthan D. T. Samuel,et al.  Optogenetic manipulation of neural activity in freely moving Caenorhabditis elegans , 2011, Nature Methods.

[13]  L B Cohen,et al.  Optical measurement of membrane potential. , 1978, Reviews of physiology, biochemistry and pharmacology.

[14]  S. Antic,et al.  Optical signals from neurons with internally applied voltage-sensitive dyes , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[15]  George J Augustine,et al.  Optogenetic mapping of cerebellar inhibitory circuitry reveals spatially biased coordination of interneurons via electrical synapses. , 2014, Cell reports.

[16]  George J. Augustine,et al.  A Genetically Encoded Ratiometric Indicator for Chloride Capturing Chloride Transients in Cultured Hippocampal Neurons , 2000, Neuron.

[17]  Y Yarom,et al.  Patches of synchronized activity in the cerebellar cortex evoked by mossy-fiber stimulation: questioning the role of parallel fibers. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Dejan Zecevic,et al.  Imaging inhibitory synaptic potentials using voltage sensitive dyes. , 2010, Biophysical journal.

[19]  H. Hellinga,et al.  Visualization of Synaptic Inhibition with an Optogenetic Sensor Developed by Cell-Free Protein Engineering Automation , 2013, The Journal of Neuroscience.

[20]  Atsushi Miyawaki,et al.  Whole-field fluorescence microscope with digital micromirror device: imaging of biological samples. , 2003, Applied optics.

[21]  Adriano B. L. Tort,et al.  OLM interneurons differentially modulate CA3 and entorhinal inputs to hippocampal CA1 neurons , 2012, Nature Neuroscience.

[22]  Toru Ishizuka,et al.  Parallel and patterned optogenetic manipulation of neurons in the brain slice using a DMD-based projector , 2013, Neuroscience Research.

[23]  George J Augustine,et al.  Imaging activity of neuronal populations with new long-wavelength voltage-sensitive dyes , 2008, Brain cell biology.

[24]  Bradley J. Baker,et al.  Wide-field and two-photon imaging of brain activity with voltage- and calcium-sensitive dyes , 2009, Philosophical Transactions of the Royal Society B: Biological Sciences.

[25]  Richard Apps,et al.  Cerebellar cortical organization: a one-map hypothesis , 2009, Nature Reviews Neuroscience.

[26]  L. Looger,et al.  Genetically encoded neural activity indicators , 2012, Current Opinion in Neurobiology.

[27]  Matt Wachowiak,et al.  Optical monitoring of neural activity using voltage-sensitive dyes. , 2002, Methods in enzymology.

[28]  D. Maclaurin,et al.  Optical recording of action potentials in mammalian neurons using a microbial rhodopsin , 2011, Nature Methods.

[29]  S. Shoham,et al.  Patterned Optical Activation of Retinal Ganglion Cells , 2007, 2007 29th Annual International Conference of the IEEE Engineering in Medicine and Biology Society.

[30]  Henrik Jörntell,et al.  Cerebellar molecular layer interneurons – computational properties and roles in learning , 2010, Trends in Neurosciences.

[31]  Karl Deisseroth,et al.  Optogenetics in Neural Systems , 2011, Neuron.

[32]  Rainer W Friedrich,et al.  High-resolution optical control of spatiotemporal neuronal activity patterns in zebrafish using a digital micromirror device , 2012, Nature Protocols.

[33]  George J. Augustine,et al.  Optogenetic probing of functional brain circuitry , 2011, Experimental physiology.

[34]  A. Konnerth,et al.  Synaptic currents in cerebellar Purkinje cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Ethan K. Scott,et al.  Optogenetic dissection of a behavioral module in the vertebrate spinal cord , 2009, Nature.

[36]  George J Augustine,et al.  Imaging synaptic inhibition in transgenic mice expressing the chloride indicator, Clomeleon , 2006, Brain cell biology.

[37]  Leslie M. Loew,et al.  Intracellular long-wavelength voltage-sensitive dyes for studying the dynamics of action potentials in axons and thin dendrites , 2007, Journal of Neuroscience Methods.

[38]  J Midtgaard,et al.  Stellate cell inhibition of Purkinje cells in the turtle cerebellum in vitro. , 1992, The Journal of physiology.

[39]  S. Palay,et al.  Cerebellar Cortex: Cytology and Organization , 1974 .

[40]  Vincent A. Pieribone,et al.  Single Action Potentials and Subthreshold Electrical Events Imaged in Neurons with a Fluorescent Protein Voltage Probe , 2012, Neuron.

[41]  R. Tsien,et al.  pHTomato: A genetically-encoded indicator that enables multiplex interrogation of synaptic activity , 2012, Nature Neuroscience.

[42]  Y. Yarom,et al.  Cerebellar on-beam and lateral inhibition: two functionally distinct circuits. , 2000, Journal of neurophysiology.

[43]  Gang Chen,et al.  Cerebellar Cortical Molecular Layer Inhibition Is Organized in Parasagittal Zones , 2006, The Journal of Neuroscience.

[44]  W. C. Hall,et al.  High-speed mapping of synaptic connectivity using photostimulation in Channelrhodopsin-2 transgenic mice , 2007, Proceedings of the National Academy of Sciences.

[45]  H. Markram,et al.  Interneurons of the neocortical inhibitory system , 2004, Nature Reviews Neuroscience.

[46]  David Willshaw,et al.  The cerebellum as a neuronal machine , 1999 .

[47]  Xiang Zhang,et al.  All optical interface for parallel, remote, and spatiotemporal control of neuronal activity. , 2007, Nano letters.