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      Neurochemistry Study of Spinal Cord in Non-Human Primate ( Sapajus Spp.)

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          Abstract

          The spinal cord is involved in local, ascending and descending neural pathways. Few studies analyzed the distribution of neuromediators in the laminae of non-human primates along all segments. The present study described the classic neuromediators in the spinal cord of the non-human primate Sapajus spp. through histochemical and immunohistochemical methods. Nicotinamide adenine dinucleotide hydrogen phosphate-diaphorase (NADPH-d) method showed neuronal somata in the intermediolateral column (IML), central cervical nucleus (CCN), laminae I, II, III, IV, V, VI, VII, VIII and X, besides dense presence of nerve fibers in laminae II and IX. Acetylcholinesterase (AChE) activity was evident in the neuronal somata in laminae V, VI, VII, VIII, IX, CCN, IML and in the Clarke’s column (CC). Immunohistochemistry data revealed neuronal nitric oxide synthase (nNOS) immunoreactivity in neuronal somata and in fibers of laminae I, II, III, VII, VIII, X and IML; choline acetyltransferase (ChAT) in neuronal somata and in fibers of laminae VII, VIII and IX; calcitonin gene-related peptide (CGRP) was noticed in neuronal somata of lamina IX and in nerve fibers of laminae I, II, III, IV, V, VI and VII; substance P (SP) in nerve fibers of laminae I, II, III, IV, V, VI, VII, VIII, IX, X, CCN, CC and IML; serotonin (5-HT) and vesicular glutamate transporter-1 (VGLUT1) was noticed in nerve fibers of all laminae; somatostatin (SOM) in neuronal somata of laminae III, IV, V, VI, VII, VIII and IX and nerve fibers in laminae I, II, V, VI, VII, X and IML; calbindin (Cb) in neuronal somata of laminae I, II, VI, VII, IX and X; parvalbumin (PV) was found in neuronal somata and in nerve fibers of laminae III, IV, V, VI, VII, VIII, IX and CC; finally, gamma-amino butyric acid (GABA) was present in neuronal somata of laminae V, VI, VII, VIII, IX and X. This study revealed interesting results concerning the chemoarchitecture of the Sapajus spp . spinal cord with a distribution pattern mostly similar to other mammals. The data corroborate the result described in literature, except for some differences in CGRP, SP, Cb, PV and GABA immunoreactivities present in neuronal somata and in nerve fibers. This could suggest certain specificity for the neurochemistry distribution in this non-human primate species, besides adding relevant data to support further studies related to processes involving spinal cord components.

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          Structure and function of the brain serotonin system.

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            The cytoarchitectonic organization of the spinal cord in the cat.

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              The expression of vesicular glutamate transporters VGLUT1 and VGLUT2 in neurochemically defined axonal populations in the rat spinal cord with emphasis on the dorsal horn.

              Two vesicular glutamate transporters, VGLUT1 and VGLUT2, have recently been identified, and it has been reported that they are expressed by largely nonoverlapping populations of glutamatergic neurons in the brain. We have used immunocytochemistry with antibodies against both transporters, together with markers for various populations of spinal neurons, in an attempt to identify glutamatergic interneurons in the dorsal horn of the mid-lumbar spinal cord of the rat. The great majority (94-100%) of nonprimary axonal boutons that contained somatostatin, substance P or neurotensin, as well as 85% of those that contained enkephalin, were VGLUT2-immunoreactive, which suggests that most dorsal horn neurons that synthesize these peptides are glutamatergic. In support of this, we found that most somatostatin- and enkephalin-containing boutons (including somatostatin-immunoreactive boutons that lacked calcitonin gene-related peptide and were therefore probably derived from local interneurons) formed synapses at which AMPA receptors were present. We also investigated VGLUT expression in central terminals of primary afferents. Myelinated afferents were identified with cholera toxin B subunit; most of those in lamina I were VGLUT2-immunoreactive, whereas all those in deeper laminae were VGLUT1-immunoreactive, and some (in laminae III-VI) appeared to contain both transporters. However, peptidergic primary afferents that contained substance P or somatostatin (most of which are unmyelinated), as well as nonpeptidergic C fibres (identified with Bandeiraea simplicifolia isolectin B4) showed low levels of VGLUT2-immunoreactivity, or were not immunoreactive with either VGLUT antibody. As all primary afferents are thought to be glutamatergic, this raises the possibility that unmyelinated afferents, most of which are nociceptors, express a different vesicular glutamate transporter.
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                Author and article information

                Journal
                Eur J Histochem
                Eur J Histochem
                EJH
                European Journal of Histochemistry : EJH
                PAGEPress Publications, Pavia, Italy
                1121-760X
                2038-8306
                15 September 2016
                10 August 2016
                : 60
                : 3
                : 2623
                Affiliations
                [1 ]School of Dentistry of Araçatuba, Basic Sciences Department, São Paulo State University (UNESP) , Araçatuba
                [2 ]Biosciences Institute of Botucatu, Anatomy Department, São Paulo State University (UNESP) , Botucatu
                [3 ]School of Medicine, Federal University of Mato Grosso do Sul (UFMS), Três Lagoas , Brazil
                Author notes
                School of Medicine, UFMS Federal University of Mato Grosso do Sul, Três Lagoas. Av. Ranulpho Marques Leal 3484, 79620-080, MS, Brazil. +55.67.33453439. andre.valerio@ 123456ufms.br

                Contributions: KRTS, AVS, study design, experimental procedures, histochemical and immunohistochemical processing, microscopic analysis, data collection, figures preparation, manuscript drafting; NOB, GWLT, manuscript drafting; JACHJ, EE, JAO, CAC, contribution to microscopic analysis, interpretation of data and manuscript drafting. All authors reviewed and contributed to the different draft versions, read and approved the final manuscript.

                Conflict of interest: the authors declare no conflict of interest.

                Article
                10.4081/ejh.2016.2623
                5062631
                27734991
                acba18de-2e68-43b3-88e1-ff1e27a6e3cf
                ©Copyright K.R. Torres-da-Silva et al.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 23 January 2016
                : 17 August 2016
                Page count
                Figures: 3, Tables: 1, Equations: 0, References: 71, Pages: 8
                Categories
                Original Paper

                Clinical chemistry
                spinal cord,immunohistochemistry,non-human primate,neuromediators
                Clinical chemistry
                spinal cord, immunohistochemistry, non-human primate, neuromediators

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