Light-evoked Somatosensory Perception of Transgenic Rats That Express Channelrhodopsin-2 in Dorsal Root Ganglion Cells

In vertebrate somatosensory systems, each mode of touch-pressure, temperature or pain is sensed by sensory endings of different dorsal root ganglion (DRG) neurons, which conducted to the specific cortical loci as nerve impulses. Therefore, direct electrical stimulation of the peripheral nerve endings causes an erroneous sensation to be conducted by the nerve. We have recently generated several transgenic lines of rat in which channelrhodopsin-2 (ChR2) transgene is driven by the Thy-1.2 promoter. In one of them, W-TChR2V4, some neurons were endowed with photosensitivity by the introduction of the ChR2 gene, coding an algal photoreceptor molecule. The DRG neurons expressing ChR2 were immunohistochemically identified using specific antibodies to the markers of mechanoreceptive or nociceptive neurons. Their peripheral nerve endings in the plantar skin as well as the central endings in the spinal cord were also examined. We identified that ChR2 is expressed in a certain population of large neurons in the DRG of W-TChR2V4. On the basis of their morphology and molecular markers, these neurons were classified as mechanoreceptive but not nociceptive. ChR2 was also distributed in their peripheral sensory nerve endings, some of which were closely associated with CK20-positive cells to form Merkel cell-neurite complexes or with S-100-positive cells to form structures like Meissner's corpuscles. These nerve endings are thus suggested to be involved in the sensing of touch. Each W-TChR2V4 rat showed a sensory-evoked behavior in response to blue LED flashes on the plantar skin. It is thus suggested that each rat acquired an unusual sensory modality of sensing blue light through the skin as touch-pressure. This light-evoked somatosensory perception should facilitate study of how the complex tactile sense emerges in the brain.

[1]  P. Bennett,et al.  Subtypes of dorsal root ganglion neurons based on different inward currents as measured by whole-cell voltage clamp , 1989, Molecular and Cellular Biochemistry.

[2]  E. McLachlan,et al.  Electrophysiological properties of neurons in intact rat dorsal root ganglia classified by conduction velocity and action potential duration. , 1996, Journal of neurophysiology.

[3]  A. Dizhoor,et al.  Ectopic Expression of a Microbial-Type Rhodopsin Restores Visual Responses in Mice with Photoreceptor Degeneration , 2006, Neuron.

[4]  Xu Zhang,et al.  The development and modulation of nociceptive circuitry , 2006, Current Opinion in Neurobiology.

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

[6]  L. Santoro,et al.  Myelinated nerve endings in human skin , 2007, Muscle & nerve.

[7]  Shiaoching Gong,et al.  A gene expression atlas of the central nervous system based on bacterial artificial chromosomes , 2003, Nature.

[8]  P. J. Waddell,et al.  Soma neurofilament immunoreactivity is related to cell size and fibre conduction velocity in rat primary sensory neurons. , 1991, The Journal of physiology.

[9]  H. Hong,et al.  Capsaicin effects on brain-derived neurotrophic factor in rat dorsal root ganglia and spinal cord. , 2000, Brain research. Molecular brain research.

[10]  Oleg A. Sineshchekov,et al.  Two rhodopsins mediate phototaxis to low- and high-intensity light in Chlamydomonas reinhardtii , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. Nathans The Evolution and Physiology of Human Color Vision Insights from Molecular Genetic Studies of Visual Pigments , 1999, Neuron.

[12]  Peter Hegemann,et al.  Algal sensory photoreceptors. , 2008, Annual review of plant biology.

[13]  H. Fukuzawa,et al.  Archaeal-type rhodopsins in Chlamydomonas: model structure and intracellular localization. , 2003, Biochemical and biophysical research communications.

[14]  R. Coggeshall Fos, nociception and the dorsal horn , 2005, Progress in Neurobiology.

[15]  Luke Campagnola,et al.  Fiber-coupled light-emitting diode for localized photostimulation of neurons expressing channelrhodopsin-2 , 2008, Journal of Neuroscience Methods.

[16]  Toru Ishizuka,et al.  Visual Properties of Transgenic Rats Harboring the Channelrhodopsin-2 Gene Regulated by the Thy-1.2 Promoter , 2009, PloS one.

[17]  KouichiC . Nakamura,et al.  Neurons in Golgi-stain-like images revealed by GFP-adenovirus infection in vivo , 2000, Neuroscience Research.

[18]  Zhi-wang Li,et al.  Modulation by adenosine of GABA‐activated current in rat dorsal root ganglion neurons , 1997, The Journal of physiology.

[19]  S. McMahon,et al.  Immunocytochemical Localization of trkA Receptors in Chemically Identified Subgroups of Adult Rat Sensory Neurons , 1995, The European journal of neuroscience.

[20]  A. Ford,et al.  Immunocytochemical localization of P2X3 purinoceptors in sensory neurons in naive rats and following neuropathic injury , 1999, Pain.

[21]  Peter Hegemann,et al.  "Vision" in single-celled algae. , 2004, News in physiological sciences : an international journal of physiology produced jointly by the International Union of Physiological Sciences and the American Physiological Society.

[22]  H. Chiel,et al.  Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopsin and green algae channelrhodopsin. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[23]  S. McMahon,et al.  Tackling Pain at the Source: New Ideas about Nociceptors , 1998, Neuron.

[24]  D. Maxwell,et al.  Spinal dorsal horn neurone targets for nociceptive primary afferents: do single neurone morphological characteristics suggest how nociceptive information is processed at the spinal level , 2004, Brain Research Reviews.

[25]  U. Kaupp,et al.  Cyclic nucleotide-gated ion channels. , 2002, Physiological reviews.

[26]  M. Gold,et al.  Inflammation-induced increase in evoked calcium transients in subpopulations of rat dorsal root ganglion neurons , 2008, Neuroscience.

[27]  D. Jacobowitz,et al.  Parvalbumin, calretinin and carbonic anhydrase in the trigeminal and spinal primary neurons of the rat , 1994, Brain Research.

[28]  C. Torsney,et al.  Characterization of Sensory Neuron Subpopulations Selectively Expressing Green Fluorescent Protein in Phosphodiesterase 1C BAC Transgenic Mice , 2006 .

[29]  R H LaMotte,et al.  Effects of a chronic compression of the dorsal root ganglion on voltage-gated Na+ and K+ currents in cutaneous afferent neurons. , 2006, Journal of neurophysiology.

[30]  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.

[31]  Z. Wiesenfeld‐Hallin,et al.  Collateral reinnervation and expansive regenerative reinnervation by sensory axons into “foreign” denervated skin: an immunohistochemical study in the rat , 2004, Experimental Brain Research.

[32]  R. North,et al.  P2X3 is expressed by DRG neurons that terminate in inner lamina II , 1998, The European journal of neuroscience.

[33]  A. Basbaum,et al.  Molecular mechanisms of nociception , 2001, Nature.

[34]  S. Waxman,et al.  Intense Isolectin-B4 Binding in Rat Dorsal Root Ganglion Neurons Distinguishes C-Fiber Nociceptors with Broad Action Potentials and High Nav1.9 Expression , 2006, The Journal of Neuroscience.

[35]  M. Zylka,et al.  Mrgprd-Expressing Polymodal Nociceptive Neurons Innervate Most Known Classes of Substantia Gelatinosa Neurons , 2009, The Journal of Neuroscience.

[36]  T. Brennan,et al.  The mammalian sodium channel BNC1 is required for normal touch sensation , 2000, Nature.

[37]  Gary R Lewin,et al.  Evidence for a protein tether involved in somatic touch , 2010, The EMBO journal.

[38]  E. Bamberg,et al.  Channelrhodopsin-1: A Light-Gated Proton Channel in Green Algae , 2002, Science.

[39]  T. Ishizuka,et al.  Kinetic evaluation of photosensitivity in genetically engineered neurons expressing green algae light-gated channels , 2006, Neuroscience Research.

[40]  K. Anderson,et al.  Patterning cell types in the dorsal spinal cord: what the mouse mutants say , 2003, Nature Reviews Neuroscience.

[41]  J. Phillips,et al.  Expression of α1-adrenoceptors on peripheral nociceptive neurons , 2011, Neuroscience.

[42]  R. Moll,et al.  Human Merkel cells--aspects of cell biology, distribution and functions. , 2005, European journal of cell biology.

[43]  Toru Ishizuka,et al.  Restoration of visual response in aged dystrophic RCS rats using AAV-mediated channelopsin-2 gene transfer. , 2007, Investigative ophthalmology & visual science.

[44]  M. Celio,et al.  Calbindin D-28k and parvalbumin in the rat nervous system , 1990, Neuroscience.

[45]  Krzysztof Palczewski,et al.  G protein-coupled receptor rhodopsin. , 2006, Annual review of biochemistry.

[46]  S. Lawson,et al.  Neurofilament immunoreactivity in populations of rat primary afferent neurons: A quantitative study of phosphorylated and non-phosphorylated subunits , 1991, Journal of neurocytology.

[47]  A. Todd,et al.  Anatomy of Primary Afferents and Projection Neurones in the Rat Spinal Dorsal Horn with Particular Emphasis on Substance P and the Neurokinin 1 Receptor , 2002, Experimental physiology.

[48]  Stephan Frings,et al.  Regulation of cyclic nucleotide-gated channels , 2005, Current Opinion in Neurobiology.

[49]  R MELZACK,et al.  The perception of pain. , 1961, Scientific American.