Background: Neuropathic pain is characterized by pain hypersensitivity to innocuous stimuli (tactile allodynia) that is nearly always resistant to known treatments such as non-steroidal antiinflammatory drugs or even opioids. It has been reported that some antidepressants are effective for treating neuropathic pain. However, the underlying molecular mechanisms are not well understood. We have recently demonstrated that blocking P2X 4 receptors in the spinal cord reverses tactile allodynia after peripheral nerve injury in rats, implying that P2X 4 receptors are a key molecule in neuropathic pain. We investigated a possible role of antidepressants as inhibitors of P2X 4 receptors and analysed their analgesic mechanism using an animal model of neuropathic pain.

Accumulating evidence indicates that purinergic P2X4 receptors (P2X4R: cation channels activated by extracellular ATP) expressed in spinal microglia are crucial for pathological chronic pain caused by nerve damage, suggesting a potential target for drug discovery. We identified NP-1815-PX (5-[3-(5-thioxo-4H-[1,2,4]oxadiazol-3-yl)phenyl]-1H-naphtho[1, 2-b][1,4]diazepine-2,4(3H,5H)-dione) as a novel antagonist selective for P2X4R with high potency and selectivity compared with other P2XR subtypes. In in vivo assay for acute and chronic pain, intrathecal administration of NP-1815-PX produced an anti-allodynic effect in mice with traumatic nerve damage without affecting acute nociceptive pain and motor function (although its oral administration did not produce the effect). Furthermore, in a mouse model of herpetic pain, P2X4R upregulation in the spinal cord exclusively occurred in microglia, and intrathecal NP-1815-PX suppressed induction of mechanical allodynia. This model also showed K+/Cl− cotransporter 2 (KCC2) downregulation, which is implicated in dorsal horn neuron hyperexcitability; this downregulation was restored by intrathecal treatment with NP-1815-PX or by interfering with brain-derived neurotrophic factor (BDNF) signaling, a P2X4R-activated microglial factor implicated in KCC2 downregulation. Taken together, the newly developed P2X4R antagonist NP-1815-PX produces anti-allodynic effects in chronic pain models without altering acute pain sensitivity, suggesting that microglial P2X4R could be an attractive target for treating chronic pain.

The University Medical Center Utrecht institutional review board approved this study, and informed consent was obtained from all subjects. The purpose of this study was to introduce and assess diffusion-weighted (DW) magnetic resonance (MR) neurography for imaging of the brachial plexus. DW MR neurographic images were displayed with a maximum intensity projection technique. DW MR neurography was evaluated in five healthy volunteers and five patients. DW MR neurography showed a long trajectory of the brachial plexus in all healthy volunteers. In all patients, DW MR neurography clearly showed the location of the disease. The proposed DW MR neurography technique can be used to obtain an overview image of the brachial plexus, with excellent conspicuity of the nerves and surrounding structures.

P2X4 receptors (P2X4Rs), a subtype of the purinergic P2X family, play important roles in regulating neuronal and glial functions in the nervous system. We have previously shown that the expression of P2X4Rs is upregulated in activated microglia after peripheral nerve injury and that activation of the receptors by extracellular ATP is crucial for maintaining nerve injury-induced pain hypersensitivity. However, the regulation of P2X4R expression on the cell surface of microglia is poorly understood. Here, we identify the CC chemokine receptor CCR2 as a regulator of P2X4R trafficking to the cell surface of microglia. In a quantitative cell surface biotinylation assay, we found that applying CCL2 or CCL12, endogenous ligands for CCR2, to primary cultured microglial cells, increased the levels of P2X4R protein on the cell surface without changing total cellular expression. This effect of CCL2 was prevented by an antagonist of CCR2. Time-lapse imaging of green fluorescent protein (GFP)-tagged P2X4R in living microglial cells showed that CCL2 stimulation increased the movement of P2X4R-GFP particles. The subcellular localization of P2X4R immunofluorescence was restricted to lysosomes around the perinuclear region. Notably, CCL2 changed the distribution of lysosomes with P2X4R immunofluorescence within microglial cells and induced release of the lysosomal enzyme β-hexosaminidase, indicating lysosomal exocytosis. Moreover, CCL2-stimulated microglia enhanced Akt phosphorylation by ATP applied extracellularly, a P2X4R-mediated response. These results indicate that CCL2 promotes expression of P2X4R protein on the cell surface of microglia through exocytosis of P2X4R-containing lysosomes, which may be a possible mechanism for pain hypersensitivity after nerve injury.

The purpose of this study was to assess the value of diffusion-weighted magnetic resonance imaging (DWI) in detecting esophageal cancer and assessing lymph-node status, compared with histopathological results. DWI was prospectively performed in 24 consecutive patients with esophageal cancer, using the diffusion-weighted whole-body imaging with background body signal suppression (DWIBS) sequence. DWIBS images were fused with T2-weighted images, and independently and blindly evaluated by three board-certified radiologists, regarding primary tumor detectability and lymph-node status. Apparent diffusion coefficients (ADCs) of the primary tumor and lymph nodes were also measured. Average primary tumor detection rate was 49.4%, average patient-based sensitivity and specificity for the detection of lymph-node metastasis were 77.8 and 55.6%, and average lymph-node group-based sensitivity and specificity were 39.4 and 92.6%. There were no interobserver differences among the three readers (P < 0.0001). Mean ADC of detected primary tumors was 1.26 +/- 0.29x10(-3) mm(2)/s. Mean ADC of metastatic lymph nodes (1.46 +/- 0.35x10(-3) mm(2)/s) was significantly higher (P < 0.0001) than that of nonmetastatic lymph nodes (1.15 +/- 0.24 mm(2)/s), but ADCs of both groups overlapped. In conclusion, this study suggests that DWI only has a limited role in detecting esophageal cancer and nodal staging.