Fragile X mental retardation protein controls ion channel expression and activity

Fragile X-associated disorders are a family of genetic conditions resulting from the partial or complete loss of fragile X mental retardation protein (FMRP). Among these disorders is fragile X syndrome, the most common cause of inherited intellectual disability and autism. FMRP is an RNA-binding protein involved in the control of local translation, which has pleiotropic effects, in particular on synaptic function. Analysis of the brain FMRP transcriptome has revealed hundreds of potential mRNA targets encoding postsynaptic and presynaptic proteins, including a number of ion channels. FMRP has been confirmed to bind voltage-gated potassium channels (Kv3.1 & Kv4.2) mRNAs and regulates their expression in somatodendritic compartments of neurons. Recent studies have uncovered a number of additional roles for FMRP besides RNA-regulation. FMRP was shown to directly interact with, and modulate, a number of ion channel complexes. The sodium-activated potassium (Slack) channel was the first ion channel shown to directly interact with FMRP; this interaction alters the single-channel properties of Slack channel. FMRP was also shown to interact with the auxiliary 4 subunit of the calcium-activated potassium (BK) channel; this interaction increases calcium-dependent activation of the BK channel. More recently, FMRP was shown to directly interact with the voltage-gated calcium channel, Cav2.2, and reduce its trafficking to the plasma membrane. Studies performed on animal models of fragile X syndrome have revealed links between modifications of ion channel activity and changes in neuronal excitability, suggesting that these modifications could contribute to the phenotypes observed in patients with fragile X-associated disorders

Effect of knockout of α2δ-1 on action potentials in mouse sensory neurons

Gene deletion of the voltage-gated calcium channel auxiliary subunit α2δ-1 has been shown previously to have a cardiovascular phenotype, and a reduction in mechano- and cold sensitivity, coupled with delayed development of neuropathic allodynia. We have also previously shown that dorsal root ganglion (DRG) neuron calcium channel currents were significantly reduced in α2δ-1 knockout mice. To extend our findings in these sensory neurons, we have examined here the properties of action potentials (APs) in DRG neurons from α2δ-1 knockout mice in comparison to their wild-type (WT) littermates, in order to dissect how the calcium channels that are affected by α2δ-1 knockout are involved in setting the duration of individual APs and their firing frequency. Our main findings are that there is reduced Ca(2+) entry on single AP stimulation, particularly in the axon proximal segment, reduced AP duration and reduced firing frequency to a 400 ms stimulation in α2δ-1 knockout neurons, consistent with the expected role of voltage-gated calcium channels in these events. Furthermore, lower intracellular Ca(2+) buffering also resulted in reduced AP duration, and a lower frequency of AP firing in WT neurons, mimicking the effect of α2δ-1 knockout. By contrast, we did not obtain any consistent evidence for the involvement of Ca(2+)-activation of large conductance calcium-activated potassium (BK) and small conductance calcium-activated potassium (SK) channels in these events. In conclusion, the reduced Ca(2+) elevation as a result of single AP stimulation is likely to result from the reduced duration of the AP in α2δ-1 knockout sensory neurons.This article is part of the themed issue 'Evolution brings Ca(2+) and ATP together to control life and death'

Analysis of Alfven Eigenmode destabilization in DIII-D high poloidal β\beta discharges using a Landau closure model

Alfven Eigenmodes are destabilized at the DIII-D pedestal during transient beta drops in high poloidal beta discharges with internal transport barriers (ITBs), driven by n=1 external kink modes, leading to energetic particle losses. There are two different scenarios in the thermal beta recovery phase: with bifurcation (two instability branches with different frequencies) or without bifurcation (single instability branch). We use the reduced MHD equations in a full 3D system, coupled with equations of density and parallel velocity moments for the energetic particles as well as the geodesic acoustic wave dynamics, to study the properties of the instabilities observed in the DIII-D high poloidal beta discharges and identify the conditions to trigger the bifurcation. The simulations suggest that instabilities with lower frequency in the bifurcation case are ballooning modes driven at the plasma pedestal, while the instability branch with higher frequencies are low n (n<4) Toroidal Alfven Eigenmodes nearby the pedestal. The reverse shear region between the middle and plasma periphery in the non-bifurcated case avoids the excitation of ballooning modes at the pedestal, although Toroidal Alfven Eigenmodes and Reverse Shear Alfven Eigenmodes are unstable in the reverse shear region. The n=1 and n=2 Alfven Eigenmode activity can be suppressed or minimized if the neutral beam injector (NBI) intensity is lower than the experimental value (betaf < 0.03). In addition, if the beam energy or neutral beam injector voltage is lower than in the experiment (Vth,f / VA0 < 0.2), the resonance between beam and thermal plasma is weaker. The n=3,4,5 and 6 AE activity can't be fully suppressed, although the growth rate and frequency is smaller for an optimized neutral beam injector operation regime. In conclusion, AE activity in high poloidal beta discharges can be minimized for optimized NBI operation regimes

Proteolytic maturation of α 2 δ represents a checkpoint for activation and neuronal trafficking of latent calcium channels

The auxiliary α2δ subunits of voltage-gated calcium channels are extracellular membrane-associated proteins, which are post-translationally cleaved into disulfide-linked polypeptides α2 and δ. We now show, using α2δ constructs containing artificial cleavage sites, that this processing is an essential step permitting voltage-dependent activation of plasma membrane N-type (CaV2.2) calcium channels. Indeed, uncleaved α2δ inhibits native calcium currents in mammalian neurons. By inducing acute cell-surface proteolytic cleavage of α2δ, voltage-dependent activation of channels is promoted, independent from the trafficking role of α2δ. Uncleaved α2δ does not support trafficking of CaV2.2 channel complexes into neuronal processes, and inhibits Ca2+ entry into synaptic boutons, and we can reverse this by controlled intracellular proteolytic cleavage. We propose a model whereby uncleaved α2δ subunits maintain immature calcium channels in an inhibited state. Proteolytic processing of α2δ then permits voltage-dependent activation of the channels, acting as a checkpoint allowing trafficking only of mature calcium channel complexes into neuronal processes

Comparative Analysis of the Frequency and Distribution of Stem and Progenitor Cells in the Adult Mouse Brain

cells (NSCs) and progenitor cells, but it cannot discriminate between these two populations. Given two assays have purported to overcome this shortfall, we performed a comparative analysis of the distribution and frequency of NSCs and progenitor cells detected in 400 m coronal segments along the ventricular neuraxis of the adult mouse brain using the neurosphere assay, the neural colony forming cell assay (N-CFCA), and label-retaining cell (LRC) approach. We observed a large variation in the number of progenitor/stem cells detected in serial sections along the neuraxis, with the number of neurosphereforming cells detected in individual 400 m sections varying from a minimum of eight to a maximum of 891 depending upon the rostral-caudal coordinate assayed. Moreover, the greatest variability occurred in the rostral portion of the lateral ventricles, thereby explaining the large variation in neurosphere frequency previously reported. Whereas the overall number of neurospheres (3730 276) or colonies (4275 124) we detected along the neuraxis did not differ significantly, LRC numbers were significantly reduced (1186 188, 7 month chase) in comparison to both total colonies and neurospheres. Moreover, approximately two orders of magnitude fewer NSC-derived colonies (50 10) were detected using the N-CFCA as compared to LRCs. Given only 5% of the LRCs are cycling (BrdU/Ki-67) or competent to divide (BrdU/Mcm-2), and proliferate upon transfer to culture, it is unclear whether this technique selectively detects endogenous NSCs. Overall, caution should be taken with the interpretation and employment of all these techniques

Forward pi^0 Production and Associated Transverse Energy Flow in Deep-Inelastic Scattering at HERA

Deep-inelastic positron-proton interactions at low values of Bjorken-x down to x \approx 4.10^-5 which give rise to high transverse momentum pi^0 mesons are studied with the H1 experiment at HERA. The inclusive cross section for pi^0 mesons produced at small angles with respect to the proton remnant (the forward region) is presented as a function of the transverse momentum and energy of the pi^0 and of the four-momentum transfer Q^2 and Bjorken-x. Measurements are also presented of the transverse energy flow in events containing a forward pi^0 meson. Hadronic final state calculations based on QCD models implementing different parton evolution schemes are confronted with the data.Comment: 27 pages, 8 figures and 3 table

The role of lubricant feeding conditions on the performance improvement and friction reduction of journal bearings

Most conventional hydrodynamic journal bearing performance tools can not suitably assess the effect of lubricant feeding conditions on bearing performance, even though these conditions are known to affect important performance parameters such as eccentricity and powerloss. A thermohydrodynamic analysis suitable to deal with realistic feeding conditions has been proposed. Special attention was given to the treatment of phenomena taking place within grooves and their vicinity,as well as to the ruptured film region. The effec to flubricant feeding pressure and temperature, groove length ratio,width ratio and number (single/twin) on bearing performance has been analyzed for a broad range of conditions.It was found that a careful tuning of the feeding conditions may indeed improve bearing performance.FCT - POCTI/EME/39202/200

The Site of the Immune Reaction Against \u3ci\u3eEimeria bovis\u3c/i\u3e in Calves and the Demonstration of Antibodies by Immunofluorescence

In 1959 Senger et al. reported that calves which had received a small oral inoculation of Eimeria bovis oocysts were later resistant to larger challenge inoculations. In an investigation on the mechanism of this immune reaction, Hammond et al. (1959) compared the number and size of E. bovis schizonts in immunized and nonimmunized calves, but found no significant difference in the results. They conducted 3 experiments using 28 young male Holstein-Friesian calves, and killed the majority of these calves for examination 17 to 20 days after the challenge inoculation