title: Spinal motoneuron synaptic plasticity after axotomy in the absence of inducible nitric oxide synthase.
abstract: BACKGROUND
  Astrocytes play a major role in preserving and restoring structural and physiological integrity following injury to the nervous system. After peripheral axotomy, reactive gliosis propagates within adjacent spinal segments, influenced by the local synthesis of nitric oxide (NO). The present work investigated the importance of inducible nitric oxide synthase (iNOS) activity in acute and late glial responses after injury and in major histocompatibility complex class I (MHC I) expression and synaptic plasticity of inputs to lesioned alpha motoneurons.
  
  
  METHODS
  In vivo analyses were carried out using C57BL/6J-iNOS knockout (iNOS(-/-)) and C57BL/6J mice. Glial response after axotomy, glial MHC I expression, and the effects of axotomy on synaptic contacts were measured using immunohistochemistry and transmission electron microscopy. For this purpose, 2-month-old animals were sacrificed and fixed one or two weeks after unilateral sciatic nerve transection, and spinal cord sections were incubated with antibodies against classical MHC I, GFAP (glial fibrillary acidic protein - an astroglial marker), Iba-1 (an ionized calcium binding adaptor protein and a microglial marker) or synaptophysin (a presynaptic terminal marker). Western blotting analysis of MHC I and nNOS expression one week after lesion were also performed. The data were analyzed using a two-tailed Student's t test for parametric data or a two-tailed Mann-Whitney U test for nonparametric data.
  
  
  RESULTS
  A statistical difference was shown with respect to astrogliosis between strains at the different time points studied. Also, MHC I expression by iNOS(-/-) microglial cells did not increase at one or two weeks after unilateral axotomy. There was a difference in synaptophysin expression reflecting synaptic elimination, in which iNOS(-/-) mice displayed a decreased number of the inputs to alpha motoneurons, in comparison to that of C57BL/6J.
  
  
  CONCLUSIONS
  The findings herein indicate that iNOS isoform activity influences MHC I expression by microglial cells one and two weeks after axotomy. This finding was associated with differences in astrogliosis, number of presynaptic terminals and synaptic covering of alpha motoneurons after lesioning in the mutant mice.
authors.affiliation: Department of Anatomy, Institute of Biology, University of Campinas (UNICAMP), CP 6109, CEP 13083-970, Campinas, SP, Brazil.
authors.forename: Amanda
authors.initials: A
authors.lastname: Emirandetti
authors.forename: Gustavo F
authors.initials: GF
authors.lastname: Simões
authors.forename: Renata G
authors.initials: RG
authors.lastname: Zanon
authors.forename: Alexandre L R
authors.initials: AL
authors.lastname: Oliveira
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000378:metabolism; D019771:Axotomy; D015415:Biomarkers / Q000378:metabolism; D005805:Genes, MHC Class I; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009046:Motor Neurons / Q000166:cytology / Q000502:physiology*; D009473:Neuronal Plasticity / Q000502:physiology*; D052248:Nitric Oxide Synthase Type I / Q000235:genetics / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism*; D017729:Presynaptic Terminals / Q000378:metabolism* / Q000648:ultrastructure; D012584:Sciatic Nerve / Q000293:injuries / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology*; D016708:Synaptophysin / Q000378:metabolism
chemical_list: D015415:Biomarkers; D007527:Isoenzymes; D016708:Synaptophysin; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; C496321:Nos1 protein, mouse; C496317:Nos2 protein, mouse