Gonadectomy induces site-specific differences in nociception in rats.

High prevalence of chronic orofacial pain in women and its relationship with ovarian states suggest that ovarian hormones may be involved in the control of orofacial nociception. Since the interaction between ovarian hormones and nociception seems more evident in the orofacial area than in many other parts of the body, a possible site specificity of an ovarian hormone effect on nociception was tested in rats. Two nociceptive tests were applied to three groups of male rats (n=46) and three groups of female rats (n=46), that were gonadectomised (n=17), sham-operated (n=15) or intact (n=14). Each rat in each group received a local subcutaneous injection of formalin in the upper lip and in the hindpaw. Upper lip injection resulted in an increased occurrence of upper lip rubbing for more than 45 min and hindpaw injection resulted in an increased occurrence of hindpaw licking for about 1h. The duration of the nociceptive behaviours was measured at 3 months after surgery. No significant difference was found between intact and sham-operated animals. A significant increase (54%) in the upper lip rubbing but not the hindpaw licking was observed in gonadectomised females. No difference was observed in castrated males for upper lip rubbing, but a tendency towards an increased duration (102%) of hindpaw licking was noted. The depletion in gonadal hormones was confirmed 3 months after gonadectomy and after the sacrifice of the animals, by the observed decline in the bone mineral density measured on the femur of 40 rats belonging to the six groups. A role of ovarian hormones was also suggested after immunostaining of oestrogen receptors in the lamina II of Caudalis subnucleus of the trigeminal sensory complex and cervical (C1-C2) spinal dorsal horn. The number of cells expressing oestrogen receptors displayed a small (13.6%) but significant (P=0.037) increase in ovariectomised compared with sham-operated rats. These results suggest that the lack of ovarian hormones induces a site-specific increase in the sensitivity to orofacial nociceptive stimulation, and that an up-regulation of oestrogen receptors in the Caudalis subnucleus and C1-C2 dorsal horn may be one of the factors involved in this effect.

Strychnine alters response properties of trigeminal nociceptive neurons in the rat.

The purpose of this study was to examine the role of glycine in sensory processes in the spinal trigeminal nucleus oralis (Sp5O). We evaluated the effect of intravenous administration of strychnine, a glycine receptor antagonist, on the responses of Sp5O convergent neurons evoked by innocuous peripheral electrical and mechanical stimuli in halothane-anesthetized rats. Strychnine significantly increased the Abeta-fiber-evoked activities of Sp5O neurons to electrical stimulation in a dose-dependent (0.2-0.8 mg/kg) fashion. The response to air-jet stimulation was also significantly enhanced at the highest dose of strychnine. These findings indicate that glycinergic neurons participate in the control of the flow of information conveyed to Sp5O nociceptive neurons by myelinated low-threshold mechanoreceptive afferents. Thus, alteration of trigeminal glycinergic modulation may contribute to the dynamic mechanical allodynia that occurs in trigeminal neuropathies.

Connexin channels in Schwann cells and the development of the X-linked form of Charcot-Marie-Tooth disease.

Charcot-Marie-Tooth disease comprises a group of genetically heterogenous disorders of the peripheral nervous system. The X-linked form of Charcot-Marie-Tooth (CMTX) is associated with mutations in the gene encoding the gap junction protein connexin32 (Cx32), which is expressed in Schwann cells. Immunocytochemical evidence suggests that Cx32 is localized to the incisures of Schmidt-Lanterman and the paranodes of myelinating Schwann cells, where it appears to form reflexive gap junctions. It is currently thought that this cytoplasmic continuity provides a much shorter diffusion pathway for the transport of ions, metabolites and second messenger molecules through intracellular channels between the adaxonal and peri-nuclear regions of Schwann cells, across the myelin sheath. This review summarizes our current understanding of the role of connexins in Schwann cells and focuses on the lessons for channel function and disease pathophysiology derived from the functional analysis of Cx32 mutations. One of the most intriguing aspects emerging from this work is that several mutations retain functional competence, although the mutated channels exhibit altered gating properties. This suggests that partial and/or selective disruption of the radial communication pathway formed by Cx32 is sufficient to cause a functional deficit and lead to the development of CMTX. The next challenge will be to define, at the molecular level, the sequence of events involved in the disease process. The presence of a group of functional mutations should help understand the cellular basis of CMTX, by allowing the identification of the specific molecules that need to be exchanged through Cx32 channels, but are excluded from the mutated ones.

Connexin32 mutations associated with X-linked Charcot-Marie-Tooth disease show two distinct behaviors: loss of function and altered gating properties.

The X-linked form of Charcot-Marie-Tooth disease (CMTX) is associated with mutations in the gene encoding connexin32 (Cx32), which is expressed in Schwann cells. We have compared the functional properties of 11 Cx32 mutations with those of the wild-type protein by testing their ability to form intercellular channels in the paired oocyte expression system. Although seven mutations were functionally incompetent, four others were able to generate intercellular currents of the same order of magnitude as those induced by wild-type Cx32 (Cx32wt). In homotypic oocyte pairs, CMTX mutations retaining functional activity induced the development of junctional currents that exhibited changes in the sensitivity and kinetics of voltage dependence with respect to that of Cx32wt. The four mutations were also capable of interacting in heterotypic configuration with the wild-type protein, and in one case the result was a marked rectification of junctional currents in response to voltage steps of opposite polarity. In addition, the functional CMTX mutations displayed the same selective pattern of compatibility as Cx32wt, interacting with Cx26, Cx46, and Cx50 but failing to do so with Cx40. Although the functional mutations exhibited sensitivity to cytoplasmic acidification, which induced a >/=80% decrease in junctional currents, both the rate and extent of channel closure were enhanced markedly for two of them. Together, these results indicate that the functional consequences of CMTX mutations of Cx32 are of two drastically distinct kinds. The presence of a functional group of mutations suggests that a selective deficit of Cx32 channels may be sufficient to impair the homeostasis of Schwann cells and lead to the development of CMTX.

New mutations in the X-linked form of Charcot-Marie-Tooth disease.

Mutations in the gene for connexin 32 are associated with a chromosome X-linked form of Charcot-Marie-Tooth disease. The prevalence of this form is probably underestimated. We screened 12 candidate families and found 7 missense mutations of which 4 are new. These mutations are located in intra- and extramembraneous parts of the protein. Some mutations are probably present with a higher frequency. This study further confirms variation of connexin 32 mutations with scarcity in the second transmembrane domain and, so far, absence in the fourth transmembrane domain and in the carboxy-terminal region.

Connexins, gap junctions and cell-cell signalling in the nervous system.

Connexins form a multigene family of polytopic membrane proteins that, in vertebrates, are the constitutive subunits of intercellular channels and provide the structural basis for electrical coupling. The appearance of electrical coupling in the nervous system is developmentally regulated and restricted to distinct cell types. Electrical coupling between neurons persists after the establishment of chemical transmission, thus suggesting that this form of cell-cell signalling may be functionally interrelated with, rather than alternative to chemical transmission. Furthermore, evidence for the possible role of gap junctions in human neurological diseases is also mounting, following the discovery that the X-linked form of Charcot-Marie-Tooth syndrome, a demyelinating neuropathy of the peripheral nervous system, is associated with mutations in a connexin gene. These findings raise new questions on the significance of connexin diversity and on their functional role in the nervous system.

X-linked dominant Charcot-Marie-Tooth neuropathy (CMTX): new mutations in the connexin32 gene.

X-linked dominant Charcot-Marie-Tooth (CMTX) neuropathy has been mapped to the Xq13 region. Subsequently, several mutations that could account for CMTX have been detected in the coding part of the connexin32 (Cx32) gene, which is located within this region. In order to develop more specific diagnostic tools, we have begun a systematic screening of families with dominant CMTX for mutations in the coding region of the Cx32 gene. This report describes a study of ten families and different mutations segregating with the disease were detected in five of them. In addition to the previously reported Arg22stop and Arg215Trp substitutions, three novel mutations are described, including two different missense mutations at codon Arg22 (Arg22Pro and Arg22Gly), and a nonsense mutation at codon Trp133. The identification of new CMTX-causing mutations is a critical step for carrier detection and presymptomatic diagnosis, and should provide essential information on the structure-function relationship of Cx32 in vitro as well as in vivo.