Substantial evidence shows that activation of glial cells in the spinal cord may promote central sensitization and enhancement of pain. Descending facilitation from the rostroventromedial medulla (RVM) is also recognized as a critical component in the maintenance of chronic pain states, although the precise mechanisms driving this activity are unclear. Here, we investigated the possibility that glial activation in the RVM could promote descending facilitation from the RVM in states of enhanced pain. Peripheral inflammation was induced with carrageenan injected into the plantar aspect of the hindpaw of male Sprague-Dawley rats and behavioral responses to noxious thermal and light tactile stimuli were determined. Microinjection of the glial inhibitors minocycline or fluorocitrate, or of SB 203580, a p38 MAPK inhibitor, produced a significant and time-related reversal of behavioral hypersensitivity resulting from hindpaw inflammation. Moreover, carrageenan-induced inflammation appeared to produce an increase in immunolabeling for activated microglia and astrocytes in the RVM, as well as for phosphorylated p38 MAPK; the latter was localized to both microglia and neurons of the RVM. Microinjection of the glial inhibitors into the RVM appeared to diminish immunofluorescent labeling for activated RVM microglia and astrocytes. Carrageenan-induced inflammation also increased RVM protein levels of Iba1 and GFAP and administration of minocycline or fluorocitrate into the RVM attenuated this effect. To examine a possible mechanism of glial activation, α, β-methylene-ATP was microinjected into the RVM, inducing thermal hyperalgesia, and pre-treatment with the P2X antagonists, PPADS and TNP-ATP, delayed the initiation of ATP-induced hyperalgesia. Post-treatment with the antagonists had no effect on established ATP-induced or carrageenan-induced hypersensitivity. The activation of P2X receptors initiates a signaling cascade leading to the production and release of nociceptive mediators, including BDNF. The RVM microinjection of an anti- BDNF antibody reversed carrageenan-induced thermal hyperalgesia. A model of morphine-induced paradoxical pain was also used to examine the role of glial activation in the RVM. Sustained morphine administration induced tactile allodynia and RVM microinjection of minocycline, but not fluorocitrate, attenuated the behavioral hypersensitivity. Sustained morphine also induced morphological changes in microglia of the RVM, suggesting microglial activation. A third model of enhanced pain used to study medullary glial activation was the spinal nerve ligation (SNL) model of neuropathic pain. The SNL injury induced astrocyte activation within the RVM and microinjection of the astrocyte inhibitor fluorocitrate attenuated the nerve injury-induced tactile allodynia. Minocycline administered to the RVM did not attenuate the behavioral hypersensitivity, suggesting a role for astrocytes, not microglia, in nerve injury-induced enhanced pain. The data show that inflammatory, opioid-induced and neuropathic pain is associated with glial activation in the RVM which likely participates in driving descending pain facilitation via glial-neuronal communication. These findings reveal a novel site of glial modulation of pain.

Le traumatisme crânien léger (TCL) est l'un des troubles neurologiques les plus courants affectant la santé publique. Aussi, les troubles du sommeil sont fréquents chez les patients atteints de TCL. Les études chez les rongeurs montrent que certains marqueurs de plasticité synaptique diminuent après le TCL, ce qui pourrait nuire à la plasticité du cerveau. Nous suggérons que la perte de sommeil intensifie l'effet négatif de TCL, qui peut refléter les changements des marqueurs de plasticité synaptique ou des changements des voies physiologiques qui régulent le sommeil. En utilisant un modèle de traumatisme crânien sur crâne fermé (closed head injury), nous avons étudié la relation bidirectionnelle entre le TCL et le sommeil en évaluant les effets de TCL sur l’activité électrique du cerveau par électroencéphalographie (EEG), et ceux de la privation de sommeil (PS) sur l'expression génique post-TCL. Premièrement, l'activité EEG a été enregistrée pour voir si l'architecture du sommeil est altérée suite au TCL. Nous avons ensuite voulu tester si la PS suite TCL induit des changements dans l'expression des gènes : Arc, Homer1a, Hif1a, Bdnf, Fos et éphrines, qui ont été liés à la plasticité synaptique et à la régulation du sommeil. Nous avons également étudié l'effet de la PS post-TCL sur le génome complet dans les régions cibles (cortex et l'hippocampe). Les principaux résultats obtenus dans cette étude confirment que TCL modifie de manière significative l'activité spectrale pendant l'éveil, le sommeil Rapid Eye Movement (REM) et le sommeil non-REM dans le deuxième 24 heures post-TCL. Fait intéressant, la capacité de maintenir de longues périodes d'éveil a été altérée immédiatement après TCL (première 24h post-TCL). La dynamique de l'activité delta pendant l'éveil a été modifié par le TCL. Parallèlement à ces modifications, des changements dans l'expression des gènes ont été observés dans le cortex et l'hippocampe. Seulement Arc et EfnA3 ont montré une interaction TCL / PS et ce dans l’hippocampe, tandis que l'expression de tous les autres gènes semblait être affectée par la PS ou TCL indépendamment. Nos résultats montrent pour la première fois que le TCL induit l'expression de deux chimiokines (Ccl3 et Cxcl5) à la fois dans le cortex cérébral et l'hippocampe 2,5 jours post-TCL. Également, nous avons observé que le TCL induit une diminution de l'expression de Lgals3 et S100A8 dans le cortex, et une augmentation d’Olig2 dans l'hippocampe. Les résultats concernant les effets de la PS sur le génome complet du cortex et de l'hippocampe montrent des changements significatifs dans les gènes impliqués dans diverses fonctions physiologiques, telles que les rythmes circadiens, la réponse inflammatoire, ainsi que de l'activation des cellules gliales. En général, nos résultats précisent les changements dans la qualité de l’éveil ainsi que dans l'expression de divers gènes après TCL. / Mild traumatic brain injury (mTBI) is one of the most common neurological disorders affecting public health. Sleep disorders are common in patients with mTBI. Studies in rodents show that some synaptic plasticity markers decreased after mTBI which could impair brain plasticity. We suggest that sleep loss intensifies the negative effect of mTBI, which may reflect changes of synaptic plasticity markers or changes of different physiological pathway that regulates the sleep process. Using a "closed head injury" model, we have studied the bidirectional relationship between mTBI and sleep by investigating the effects of mTBI on sleep structure, and that of sleep deprivation (SD) on gene expression post-mTBI. First, EEG activity was monitored to investigate if sleep architecture is altered following mTBI. We then tested if SD, following mTBI, induces changes in gene expression of plasticity markers (Arc, Homer1a, Hif1a, Bdnf, Fos, and Ephrins), which have also been linked to sleep regulation. We also investigated the effect of SD post-mTBI on genome wide gene expression in target regions. The main results obtained in this study confirm that mTBI affects wakefulness, and significantly changes spectral activity during wakefulness, rapid eye movement (REM) sleep, and non-REM sleep on the second 24 hours post-TCL. Interestingly, the capacity to sustain long bouts of wakefulness was impaired immediately after mTBI. In addition, delta activity time course was altered by mTBI during wakefulness. In parallel to these alterations, changes in gene expression were observed. Only Arc and EfnA3 showed a mTBI/SD interaction in the hippocampus specifically, whereas expression of all other genes seemed to be affected by SD or mTBI independently. Our results indicate for the first time that the TCL induced the expression of two chemokines (Ccl3 and Cxcl5) in the cerebral cortex and hippocampus 2.5 days post-TCL. Also, we observed that the TCL induces a decrease in the expression of Lgals3 and S100A8 in the cortex, and an increase of Olig2 in the hippocampus.Results of SD effects on genome wide gene expression in the cortex and hippocampus show significant changes in genes involved in various physiological functions, such as circadian rhythms, inflammation, and also glial cell activation. In general, our results precise changes in wakefulness as well as in expression of various genes after mTBI.