Inhibition of the PKCγ-ε Pathway Relieves from Meningeal Nociception in an Animal Model: An Innovative Perspective for Migraine Therapy?

There is convincing evidence that nitric oxide (NO) may be a causative factor in the pathogenesis of migraine. We investigated the consequences of NO donors’ administration on meningeal processes related to the development of migraine pain in an animal model of meningeal nociception. The administration in mice of the NO donors nitroglycerin (GTN) and sodium nitroprusside (SNP) produced a delayed meningeal upregulation of interleukin-1ß and inducible NO synthase. A thermal allodynia and hyperalgesia devoid of side effects was produced 1 to 4 h after administration. To clarify the cellular pathways modulated by GTN and SNP, we examined the expression of cellular factors involved in pain modulation, such as protein kinase C (PKC) and its downstream effectors. Western blotting experiments showed an upregulation and increased phosphorylation of PKCγ and PKCε within dura mater after NO donors’ administration. A dramatic PKC-dependent increase of the phosphorylation of cyclic AMP response element binding protein (CREB) and signal transducer and activator of transcription (STAT)-1 was observed, along with an activation of the nuclear factor-κB (NF-κB) pathway, as reflected by a reduction of the inhibitory protein-κ-Bα (IκBα). Furthermore, the PKC blocker, Calphostin C, prevented the GTN and SNP-induced pain hypersensitivity. These results suggest the relevance of the PKC-mediated pathway in the induction of meningeal nociception and might help clarify the etiopathology of migraines. We can suggest PKC as a new target for migraine pain. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s13311-012-0151-8) contains supplementary material, which is available to authorized users

PKC-Dependent Signaling Pathways within PAG and Thalamus Contribute to the Nitric Oxide-Induced Nociceptive Behavior

Nitric oxide (NO) is an important molecule involved in nociceptive processing in the central nervous system. The release of NO within the spinal cord has long been implicated in the mechanisms underlying exaggerated pain sensitivity, and administration of NO donors can induce hyperalgesia. To elucidate the supraspinal mechanism responsible for NO-induced nociceptive hypersensitivity, we investigated the modulation of protein kinase C (PKC) and downstream effectors following treatment with the NO donors nitroglycerin and sodium nitroprusside. Both compounds induced a prolonged cold allodynia and heat hyperalgesia, increased levels of c-Fos and IL-1β, and activated NF-κB within periaqueductal grey matter and thalamus. Simultaneously, an increased expression and phosphorylation of PKC γ and ε were detected. To clarify the cellular mechanism involved in the NO-induced hypernociception, we examined the expression of transcription factors that act as PKC downstream effectors. A dramatic hyperphosphorylation of CREB and STAT1 was observed. The i.c.v. administration of the PKC blocker calphostin C prevented the NO-induced hypernociception, the hyperphosphorylation of CREB and STAT1, and partially reduced NF-κB activation. Conversely, the increase of IL-1β was unmodified by calphostin C. These results suggest the relevance of cerebral PKC-mediated CREB and STAT1 activation in the NO donor-induced nociceptive behavior

Regionally selective activation and differential regulation of ERK, JNK and p38 MAP kinase signalling pathway by protein kinase C in mood modulation.

A growing body of evidence indicates that the extracellular signal-regulated kinase (ERK) pathway may participate in the neuronal modulation of depression. p38MAPK and c-Jun-N-terminal kinase/stress-activated protein kinase (JNK/SAPK) also belong to the MAPK family which mainly function as mediators of cellular stresses. Since increasing evidence implicates stress as an important factor in vulnerability to depressive illnesses, the involvement of ERK, JNK and p38MAPK pathways in the modulation of mood was investigated in the forced swim test (FST) and tail suspension test (TST). The effect produced by a single acute session of FST and TST on hippocampal and cortical MAPK expression and phosphorylation was investigated by immunoblotting experiments. In the hippocampus of animals exposed to FST and TST, an intensive, PKC-dependent, ERK1, ERK2, JNK, and p38MAPK phosphorylation was observed. In the frontal cortex, the FST and TST produced a PKC-dependent increase of ERK2 and p38MAPK phosphorylation, a PKC-independent activation of JNK and cAMP response element-binding protein (CREB) whereas any involvement of ERK1 was detected. The PKC blocker calphostin C (0.05-0.1 μg i.c.v.), the MEK inhibitor U0126 (10-20 μg i.c.v.), the p38MAPK inhibitor SB203580 (5-20 μg i.c.v.) and the JNK inhibitor II (0.5-5 μg i.c.v.), produced antidepressant-like behaviour without altering locomotor activity. These results illustrate a differentially mediated activation of MAPK in hippocampus and frontal cortex of animals exposed to behavioural despair paradigms. An antidepressant-like phenotype produced by acute blockade of MAPK signalling was also demonstrated