title: Innocuous, not noxious, input activates PKCgamma interneurons of the spinal dorsal horn via myelinated afferent fibers.
abstract: Protein kinase C gamma (PKCgamma), which is concentrated in interneurons of the inner part of lamina II of the dorsal horn, has been implicated in injury-induced allodynia, a condition wherein pain is produced by innocuous stimuli. Although it is generally assumed that these interneurons receive input from the nonpeptidergic, IB4-positive subset of nociceptors, the fact that PKCgamma cells do not express Fos in response to noxious stimulation suggests otherwise. Here, we demonstrate that the terminal field of the nonpeptidergic population of nociceptors, in fact, lies dorsal to that of PKCgamma interneurons. There was also no overlap between the PKCgamma-expressing interneurons and the transganglionic tracer wheat germ agglutinin which, after sciatic nerve injection, labels all unmyelinated nociceptors. However, transganglionic transport of the beta-subunit of cholera toxin, which marks the medium-diameter and large-diameter myelinated afferents that transmit non-noxious information, revealed extensive overlap with the layer of PKCgamma interneurons. Furthermore, expression of a transneuronal tracer in myelinated afferents resulted in labeling of PKCgamma interneurons. Light and electron microscopic double labeling further showed that the VGLUT1 subtype of vesicular glutamate transmitter, which is expressed in myelinated afferents, marks synapses that are presynaptic to the PKCgamma interneurons. Finally, we demonstrate that a continuous non-noxious input, generated by walking on a rotarod, induces Fos in the PKCgamma interneurons. These results establish that PKCgamma interneurons are activated by myelinated afferents that respond to innocuous stimuli, which suggests that injury-induced mechanical allodynia is transmitted through a circuit that involves PKCgamma interneurons and non-nociceptive, VGLUT1-expressing myelinated primary afferents.
authors.affiliation: Department of Anatomy and W M Keck Foundation Center for Integrative Neuroscience, University of California, San Francisco, San Francisco, California 94158, USA. neumann@phy.ucsf.edu
authors.forename: Simona
authors.initials: S
authors.lastname: Neumann
authors.forename: Joao M
authors.initials: JM
authors.lastname: Braz
authors.forename: Kate
authors.initials: K
authors.lastname: Skinner
authors.forename: Ida J
authors.initials: IJ
authors.lastname: Llewellyn-Smith
authors.forename: Allan I
authors.initials: AI
authors.lastname: Basbaum
mesh_terms: D000818:Animals; D002772:Cholera Toxin; D005557:Formaldehyde / Q000008:administration & dosage / Q000494:pharmacology; D006614:Hindlimb; D006941:Hyperesthesia / Q000503:physiopathology; D007267:Injections; D007395:Interneurons / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D009043:Motor Activity / Q000502:physiology; D009413:Nerve Fibers, Myelinated / Q000502:physiology*; D009475:Neurons, Afferent / Q000502:physiology*; D009619:Nociceptors / Q000187:drug effects / Q000502:physiology; D017729:Presynaptic Terminals / Q000378:metabolism / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism*; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D050598:Vesicular Glutamate Transport Protein 1 / Q000378:metabolism; D014909:Wheat Germ Agglutinins
chemical_list: D016760:Proto-Oncogene Proteins c-fos; D050598:Vesicular Glutamate Transport Protein 1; D014909:Wheat Germ Agglutinins; D005557:Formaldehyde; D002772:Cholera Toxin; C087857:protein kinase C gamma; D011493:Protein Kinase C