title: Nociceptive neuropeptide increases and periorbital allodynia in a model of traumatic brain injury.
abstract: OBJECTIVE
  This study tests the hypothesis that injury to the somatosensory cortex is associated with periorbital allodynia and increases in nociceptive neuropeptides in the brainstem in a mouse model of controlled cortical impact (CCI) injury.
  
  
  METHODS
  Male C57BL/6 mice received either CCI or craniotomy-only followed by weekly periorbital von Frey (mechanical) sensory testing for up to 28 days post-injury. Mice receiving an incision only and naïve mice were included as control groups. Changes in calcitonin gene-related peptide (CGRP) and substance P (SP) within the brainstem were determined using enzyme-linked immunosorbent assay and immunohistochemistry, respectively. Activation of ionized calcium-binding adaptor molecule-1-labeled macrophages/microglia and glial fibrillary acidic protein (GFAP)-positive astrocytes were evaluated using immunohistochemistry because of their potential involvement in nociceptor sensitization.
  
  
  RESULTS
  Incision-only control mice showed no changes from baseline periorbital von Frey mechanical thresholds. CCI significantly reduced mean periorbital von Frey thresholds (periorbital allodynia) compared with baseline and craniotomy-only at each endpoint, analysis of variance P < .0001. Craniotomy significantly reduced periorbital threshold at 14 days but not 7, 21, or 28 days compared with baseline threshold, P < .01. CCI significantly increased SP immunoreactivity in the brainstem at 7 and 14 days but not 28 days compared with craniotomy-only and controls, P < .001. CGRP levels in brainstem tissues were significantly increased in CCI groups compared with controls (incision-only and naïve mice) or craniotomy-only mice at each endpoint examined, P < .0001. There was a significant correlation between CGRP and periorbital allodynia (P < .0001, r = -0.65) but not for SP (r = 0.20). CCI significantly increased the number of macrophage/microglia in the injured cortex at each endpoint up to 28 days, although cell numbers declined over weeks post-injury, P < .001. GFAP(+) immunoreactivity was significantly increased at 7 but not 14 or 28 days after CCI, P < .001. Craniotomy resulted in transient periorbital allodynia accompanied by transient increases in SP, CGRP, and GFAP immunoreactivity compared with control mice. There was no increase in the number of macrophage/microglia cells compared with controls after craniotomy.
  
  
  CONCLUSIONS
  Injury to the somatosensory cortex results in persistent periorbital allodynia and increases in brainstem nociceptive neuropeptides. Findings suggest that persistent allodynia and increased neuropeptides are maintained by mechanisms other than activation of macrophage/microglia or astrocyte in the injured somatosensory cortex.
authors.affiliation: Department of Neurological Surgery, Thomas Jefferson University, 1025 Walnut Street, Philadelphia, PA 19107, USA. Melanie.elliott@jefferson.edu
authors.forename: Melanie B
authors.initials: MB
authors.lastname: Elliott
authors.forename: Michael L
authors.initials: ML
authors.lastname: Oshinsky
authors.forename: Peter S
authors.initials: PS
authors.lastname: Amenta
authors.forename: Olatilewa O
authors.initials: OO
authors.lastname: Awe
authors.forename: Jack I
authors.initials: JI
authors.lastname: Jallo
mesh_terms: D000818:Animals; D001930:Brain Injuries / Q000150:complications* / Q000378:metabolism; D001933:Brain Stem / Q000737:chemistry / Q000378:metabolism; D015740:Calcitonin Gene-Related Peptide / Q000096:biosynthesis; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D006261:Headache / Q000209:etiology* / Q000378:metabolism; D006930:Hyperalgesia / Q000209:etiology* / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009479:Neuropeptides / Q000096:biosynthesis*; D059225:Nociception / Q000502:physiology; D013003:Somatosensory Cortex / Q000293:injuries; D013373:Substance P / Q000096:biosynthesis
chemical_list: D009479:Neuropeptides; D013373:Substance P; D015740:Calcitonin Gene-Related Peptide