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      Quantitative Study of NPY-Expressing GABAergic Neurons and Axons in Rat Spinal Dorsal Horn*

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          Abstract

          Between 25–40% of neurons in laminae I–III are GABAergic, and some of these express neuropeptide Y (NPY). We previously reported that NPY-immunoreactive axons form numerous synapses on lamina III projection neurons that possess the neurokinin 1 receptor (NK1r). The aims of this study were to determine the proportion of neurons and GABAergic boutons in this region that contain NPY, and to look for evidence that they selectively innervate different neuronal populations. We found that 4–6% of neurons in laminae I–III were NPY-immunoreactive and based on the proportions of neurons that are GABAergic, we estimate that NPY is expressed by 18% of inhibitory interneurons in laminae I–II and 9% of those in lamina III. GABAergic boutons were identified by the presence of the vesicular GABA transporter (VGAT) and NPY was found in 13–15% of VGAT-immunoreactive boutons in laminae I–II, and 5% of those in lamina III. For both the lamina III NK1r-immunoreactive projection neurons and protein kinase Cγ (PKCγ)-immunoreactive interneurons in lamina II, we found that around one-third of the VGAT boutons that contacted them were NPY-immunoreactive. However, based on differences in the sizes of these boutons and the strength of their NPY-immunoreactivity, we conclude that these originate from different populations of interneurons. Only 6% of VGAT boutons presynaptic to large lamina I projection neurons that lacked NK1rs contained NPY. These results show that NPY-containing neurons make up a considerable proportion of the inhibitory interneurons in laminae I–III, and that their axons preferentially target certain classes of dorsal horn neuron. J. Comp. Neurol. 519:1007–1023, 2011. © 2010 Wiley-Liss, Inc.

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          NeuN, a neuronal specific nuclear protein in vertebrates.

          A battery of monoclonal antibodies (mAbs) against brain cell nuclei has been generated by repeated immunizations. One of these, mAb A60, recognizes a vertebrate nervous system- and neuron-specific nuclear protein that we have named NeuN (Neuronal Nuclei). The expression of NeuN is observed in most neuronal cell types throughout the nervous system of adult mice. However, some major cell types appear devoid of immunoreactivity including cerebellar Purkinje cells, olfactory bulb mitral cells, and retinal photoreceptor cells. NeuN can also be detected in neurons in primary cerebellar cultures and in retinoic acid-stimulated P19 embryonal carcinoma cells. Immunohistochemically detectable NeuN protein first appears at developmental timepoints which correspond with the withdrawal of the neuron from the cell cycle and/or with the initiation of terminal differentiation of the neuron. NeuN is a soluble nuclear protein, appears as 3 bands (46-48 x 10(3) M(r)) on immunoblots, and binds to DNA in vitro. The mAb crossreacts immunohistochemically with nervous tissue from rats, chicks, humans, and salamanders. This mAb and the protein recognized by it serve as an excellent marker for neurons in the central and peripheral nervous systems in both the embryo and adult, and the protein may be important in the determination of neuronal phenotype.
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            GlyR alpha3: an essential target for spinal PGE2-mediated inflammatory pain sensitization.

            Prostaglandin E2 (PGE2) is a crucial mediator of inflammatory pain sensitization. Here, we demonstrate that inhibition of a specific glycine receptor subtype (GlyR alpha3) by PGE2-induced receptor phosphorylation underlies central inflammatory pain sensitization. We show that GlyR alpha3 is distinctly expressed in superficial layers of the spinal cord dorsal horn. Mice deficient in GlyR alpha3 not only lack the inhibition of glycinergic neurotransmission by PGE2 seen in wild-type mice but also show a reduction in pain sensitization induced by spinal PGE2 injection or peripheral inflammation. Thus, GlyR alpha3 may provide a previously unrecognized molecular target in pain therapy.
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              The cytoarchitectonic organization of the spinal cord in the cat.

              B REXED (1952)
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                Author and article information

                Journal
                J Comp Neurol
                cne
                The Journal of Comparative Neurology
                Wiley Subscription Services, Inc., A Wiley Company
                0021-9967
                1096-9861
                15 April 2011
                23 December 2010
                : 519
                : 6
                : 1007-1023
                Affiliations
                [1 ]simpleInstitute of Neuroscience and Psychology, University of Glasgow Glasgow, G12 8QQ, UK
                [2 ]simpleDepartment of Anatomy, Hokkaido University School of Medicine Sapporo 060-8638, Japan
                Author notes
                *CORRESPONDENCE TO: Andrew J. Todd, Spinal Cord Group, West Medical Building, University of Glasgow, Glasgow, G12 8QQ, UK. E-mail: a.todd@ 123456bio.gla.ac.uk

                Grant sponsor: Wellcome Trust; Grant number: 076976.

                Re-use of this article is permitted in accordance with the Terms and Conditions set out at http://wileyonlinelibrary.com/onlineopen#OnlineOpen_Terms

                Article
                10.1002/cne.22570
                3258544
                21344400
                017409b9-7835-46f9-affe-32d53a151950
                Copyright © 2011 Wiley-Liss, Inc., A Wiley Company

                Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.

                History
                : 16 June 2010
                : 08 October 2010
                : 08 December 2010
                Categories
                Research Article

                Neurology
                neurokinin 1 receptor,gephyrin,projection neuron,inhibitory interneuron,pkcγ,confocal microscopy

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