A single serine residue confers tetrodotoxin insensitivity on the rat sensory-neuron-specific sodium channel SNS

AbstractSensory neurons express a sodium channel (SNS) that is highly resistant to block by tetrodotoxin (IC50=60 μM). SNS is 65% homologous to the cardiac sodium channel, in which a single hydrophilic residue in the SS2 segment is critical for tetrodotoxin resistance. By site-directed mutagenesis, we have substituted phenylalanine for serine at the equivalent position in SNS: this mutated (S356F) SNS channel is functionally similar to wild-type SNS when expressed in Xenopus oocytes, but is potently blocked by tetrodotoxin and saxitoxin with IC50s of 2.8 nM and 8.2 nM, respectively. These data provide clues to the rational design of selective blockers of SNS with potential as analgesic drugs

Proteomic profiling of neuromas reveals alterations in protein composition and local protein synthesis in hyper-excitable nerves

Neuropathic pain may arise following peripheral nerve injury though the molecular mechanisms associated with this are unclear. We used proteomic profiling to examine changes in protein expression associated with the formation of hyper-excitable neuromas derived from rodent saphenous nerves. A two-dimensional difference gel electrophoresis ( 2D-DIGE) profiling strategy was employed to examine protein expression changes between developing neuromas and normal nerves in whole tissue lysates. We found around 200 proteins which displayed a > 1.75-fold change in expression between neuroma and normal nerve and identified 55 of these proteins using mass spectrometry. We also used immunoblotting to examine the expression of low-abundance ion channels Nav1.3, Nav1.8 and calcium channel alpha 2 delta-1 subunit in this model, since they have previously been implicated in neuronal hyperexcitability associated with neuropathic pain. Finally, S(35)methionine in vitro labelling of neuroma and control samples was used to demonstrate local protein synthesis of neuron-specific genes. A number of cytoskeletal proteins, enzymes and proteins associated with oxidative stress were up-regulated in neuromas, whilst overall levels of voltage-gated ion channel proteins were unaffected. We conclude that altered mRNA levels reported in the somata of damaged DRG neurons do not necessarily reflect levels of altered proteins in hyper-excitable damaged nerve endings. An altered repertoire of protein expression, local protein synthesis and topological re-arrangements of ion channels may all play important roles in neuroma hyper-excitability

Neutrino oscillation parameters from MINOS, ICARUS and OPERA combined

We perform a detailed analysis of the capabilities of the MINOS, ICARUS and OPERA experiments to measure neutrino oscillation parameters at the atmospheric scale with their data taken separately and in combination. MINOS will determine $\Delta m^2_{32}$ and $\sin^2 2\theta_{23}$ to within 10% at the 99% C.L. with 10 kton-years of data. While no one experiment will determine $\sin^2 2\theta_{13}$ with much precision, if its value lies in the combined sensitivity region of the three experiments, it will be possible to place a lower bound of O(0.01) at the 95% C.L. on this parameter by combining the data from the three experiments. The same bound can be placed with a combination of MINOS and ICARUS data alone.Comment: Version to appear in PR

Na+ current properties in islet α- and β-cells reflect cell-specific Scn3a and Scn9a expression

Key points α‐ and β‐cells express both Nav1.3 and Nav1.7 Na+ channels but in different relative amounts. The differential expression explains the different properties of Na+ currents in α‐ and β‐cells. Nav1.3 is the functionally important Na+ channel α subunit in both α‐ and β‐cells. Islet Nav1.7 channels are locked in an inactive state due to an islet cell‐specific factor. Mouse pancreatic β‐ and α‐cells are equipped with voltage‐gated Na+ currents that inactivate over widely different membrane potentials (half‐maximal inactivation (V0.5) at −100 mV and −50 mV in β‐ and α‐cells, respectively). Single‐cell PCR analyses show that both α‐ and β‐cells have Nav1.3 (Scn3) and Nav1.7 (Scn9a) α subunits, but their relative proportions differ: β‐cells principally express Nav1.7 and α‐cells Nav1.3. In α‐cells, genetically ablating Scn3a reduces the Na+ current by 80%. In β‐cells, knockout of Scn9a lowers the Na+ current by >85%, unveiling a small Scn3a‐dependent component. Glucagon and insulin secretion are inhibited in Scn3a−/− islets but unaffected in Scn9a‐deficient islets. Thus, Nav1.3 is the functionally important Na+ channel α subunit in both α‐ and β‐cells because Nav1.7 is largely inactive at physiological membrane potentials due to its unusually negative voltage dependence of inactivation. Interestingly, the Nav1.7 sequence in brain and islets is identical and yet the V0.5 for inactivation is >30 mV more negative in β‐cells. This may indicate the presence of an intracellular factor that modulates the voltage dependence of inactivation

Inhibition of somatosensory mechanotransduction by annexin A6

Mechanically activated, slowly adapting currents in sensory neurons have been linked to noxious mechanosensation. The conotoxin NMB-1 (noxious mechanosensation blocker-1) blocks such currents and inhibits mechanical pain. Using a biotinylated form of NMB-1 in mass spectrometry analysis, we identified 67 binding proteins in sensory neurons and a sensory neuron-derived cell line, of which the top candidate was annexin A6, a membrane-associated calcium-binding protein. Annexin A6-deficient mice showed increased sensitivity to mechanical stimuli. Sensory neurons from these mice showed increased activity of the cation channel Piezo2, which mediates a rapidly adapting mechano-gated current linked to proprioception and touch, and a decrease in mechanically activated, slowly adapting currents. Conversely, overexpression of annexin A6 in sensory neurons inhibited rapidly adapting currents that were partially mediated by Piezo2. Furthermore, overexpression of annexin A6 in sensory neurons attenuated mechanical pain in a mouse model of osteoarthritis, a disease in which mechanically evoked pain is particularly problematic. These data suggest that annexin A6 can be exploited to inhibit chronic mechanical pain

High Involvement Management, High Performance Work Systems and Well-being

Studies on the impact of high-performance work systems on employees' well-being are emerging but the underlying theory remains weak. This paper attempts to develop theory of the effects on well-being of four dimensions of high-performance work systems: enriched jobs, high involvement management, employee voice, and motivational supports. Hypothesized associations are tested using multilevel models and data from Britain's Workplace Employment Relations Survey of 2004 (WERS2004). Results show that enriched jobs are positively associated with both measures of well-being: job satisfaction and anxiety–contentment. Voice is positively associated with job satisfaction, and motivational supports with neither measure. The results for high involvement management are not as predicted because it increases anxiety and is independent of job satisfaction

The Large Magellanic Cloud and the Distance Scale

The Magellanic Clouds, especially the Large Magellanic Cloud, are places where multiple distance indicators can be compared with each other in a straight-forward manner at considerable precision. We here review the distances derived from Cepheids, Red Variables, RR Lyraes, Red Clump Stars and Eclipsing Binaries, and show that the results from these distance indicators generally agree to within their errors, and the distance modulus to the Large Magellanic Cloud appears to be defined to 3% with a mean value of 18.48 mag, corresponding to 49.7 Kpc. The utility of the Magellanic Clouds in constructing and testing the distance scale will remain as we move into the era of Gaia.Comment: 23 pages, accepted for publication in Astrophysics and Space Science. From a presentation at the conference The Fundamental Cosmic Distance Scale: State of the Art and the Gaia Perspective, Naples, May 201