Partial block by riluzole of muscle sodium channels in myotubes from amyotrophic lateral sclerosis patients.

International audienceDenervated muscles undergo fibrillations due to spontaneous activation of voltage-gated sodium (Na(+)) channels generating action potentials. Fibrillations also occur in patients with amyotrophic lateral sclerosis (ALS). Riluzole, the only approved drug for ALS treatment, blocks voltage-gated Na(+) channels, but its effects on muscle Na(+) channels and fibrillations are yet poorly characterized. Using patch-clamp technique, we studied riluzole effect on Na(+) channels in cultured myotubes from ALS patients. Needle electromyography was used to study fibrillation potentials (Fibs) in ALS patients during riluzole treatment and after one week of suspension. Patients were clinically characterized in all recording sessions. In myotubes, riluzole (1 μM, a therapeutic concentration) reduced Na(+) current by 20%. The rate of rise and amplitude of spikes evoked by depolarizing stimuli were also reduced. Fibs were detected in all patients tested during riluzole treatment and riluzole washout had no univocal effect. Our study indicates that, in human myotubes, riluzole partially blocks Na(+) currents and affects action potentials but does not prevent firing. In line with this in vitro finding, muscle Fibs in ALS patients appear to be largely unaffected by riluzole

Enriched environment reduces glioma growth through immune and non-immune mechanisms in mice

Mice exposed to standard (SE) or enriched environment (EE) were transplanted with murine or human glioma cells and differences in tumour development were evaluated. We report that EE exposure affects: (i) tumour size, increasing mice survival; (ii) glioma establishment, proliferation and invasion; (iii) microglia/macrophage (M/Mφ) activation; (iv) natural killer (NK) cell infiltration and activation; and (v) cerebral levels of IL-15 and BDNF. Direct infusion of IL-15 or BDNF in the brain of mice transplanted with glioma significantly reduces tumour growth. We demonstrate that brain infusion of IL-15 increases the frequency of NK cell infiltrating the tumour and that NK cell depletion reduces the efficacy of EE and IL-15 on tumour size and of EE on mice survival. BDNF infusion reduces M/Mφ infiltration and CD68 immunoreactivity in tumour mass and reduces glioma migration inhibiting the small G protein RhoA through the truncated TrkB.T1 receptor. These results suggest alternative approaches for glioma treatment

Cytosolic serine hydroxymethyltransferase controls lung adenocarcinoma cells migratory ability by modulating AMP kinase activity

Nutrient utilization and reshaping of metabolism in cancer cells is a well-known driver of malignant transformation. Less clear is the influence of the local microenvironment on metastasis formation and choice of the final organ to invade. Here we show that the level of the amino acid serine in the cytosol affects the migratory properties of lung adenocarcinoma (LUAD) cells. Inhibition of serine or glycine uptake from the extracellular milieu, as well as knockdown of the cytosolic one-carbon metabolism enzyme serine hydroxymethyltransferase (SHMT1), abolishes migration. Using rescue experiments with a brain extracellular extract, and direct measurements, we demonstrate that cytosolic serine starvation controls cell movement by increasing reactive oxygen species formation and decreasing ATP levels, thereby promoting activation of the AMP sensor kinase (AMPK) by phosphorylation. Activation of AMPK induces remodeling of the cytoskeleton and finally controls cell motility. These results highlight that cytosolic serine metabolism plays a key role in controlling motility, suggesting that cells are able to dynamically exploit the compartmentalization of this metabolism to adapt their metabolic needs to different cell functions (movement vs. proliferation). We propose a model to explain the relevance of serine/glycine metabolism in the preferential colonization of the brain by LUAD cells and suggest that the inhibition of serine/glycine uptake and/or cytosolic SHMT1 might represent a successful strategy to limit the formation of brain metastasis from primary tumors, a major cause of death in these patients

Microglia modulates hippocampal synaptic transmission and sleep duration along the light/dark cycle

Microglia, the brain's resident macrophages, actively contributes to the homeostasis of cerebral parenchyma by sensing neuronal activity and supporting synaptic remodeling and plasticity. While several studies demonstrated different roles for astrocytes in sleep, the contribution of microglia in the regulation of sleep/wake cycle and in the modulation of synaptic activity in the different day phases has not been deeply investigated. Using light as a zeitgeber cue, we studied the effects of microglial depletion with the colony stimulating factor-1 receptor antagonist PLX5622 on the sleep/wake cycle and on hippocampal synaptic transmission in male mice. Our data demonstrate that almost complete microglial depletion increases the duration of NREM sleep and reduces the hippocampal excitatory neurotransmission. The fractalkine receptor CX3CR1 plays a relevant role in these effects, because cx3cr1GFP/GFP mice recapitulate what found in PLX5622-treated mice. Furthermore, during the light phase, microglia express lower levels of cx3cr1 and a reduction of cx3cr1 expression is also observed when cultured microglial cells are stimulated by ATP, a purinergic molecule released during sleep. Our findings suggest that microglia participate in the regulation of sleep, adapting their cx3cr1 expression in response to the light/dark phase, and modulating synaptic activity in a phase-dependent manner.Bordeaux Region Aquitaine Initiative for Neuroscienc

Rubin-Euclid Derived Data Products:Initial Recommendations

This report is the result of a joint discussion between the Rubin and Euclid scientific communities. The work presented in this report was focused on designing and recommending an initial set of Derived Data products (DDPs) that could realize the science goals enabled by joint processing. All interested Rubin and Euclid data rights holders were invited to contribute via an online discussion forum and a series of virtual meetings. Strong interest in enhancing science with joint DDPs emerged from across a wide range of astrophysical domains: Solar System, the Galaxy, the Local Volume, from the nearby to the primaeval Universe, and cosmology

Nicotinic cholinergic stimulation promotes survival and reduces motility of cultured rat cerebellar granule cells

Despite many studies on the functional expression of neuronal nicotinic acetylcholine receptors (nAChRs), an exhaustive description of the long-term effects of nicotine (Nic) stimulation in cerebellar granules is still far to be completed. For this reason, we addressed the experiments stimulating cultured cerebellar granule neurons (CGN) with Nic, focusing on the effects on cell motility and survival. Using electrophysiological and Ca2+-fluorescence techniques, we found a subset of rat CGN that responded to Nic by inward whole cell currents and by short-delay Ca2+ transients. These responses were mediated through both homomeric and heteromeric nAChRs, as assessed by their sensitivity to alpha-bungarotoxin (alpha-BTX), dihydro-beta-erythroidine (DHbetaE), methyllicaconitine (MLA) and 5-hydroxyindole (5OH-indole). Once established the expression of alpha-BTX-sensitive and insensitive nAChRs and their ability to trigger Ca2+ responses in CGN, we aimed at investigating their possible role on cell survival and motility. We demonstrate that Nic stimulation significantly increases the survival of CGN exposed to the apoptosis-promoting low K+ medium. This anti-apoptotic effect is likely mediated through alpha7* nAChRs since we found that it was mimicked by choline, was insensitive to DHbetaE and was fully inhibited by alpha-BTX. Furthermore, we report that Nic negatively modulates CGN motility, reducing the basal cell movement through a pored membrane by the activation of alpha-BTX-insensitive nAChRs. We conclude that CGN express various types of nAChRs, which are differently involved in regulating Nic-mediated modulation of cell survival and migration, and we suggest potential regulatory roles for cholinergic receptors during cerebellar development. (C) 2004 IBRO. Published by Elsevier Ltd. All rights reserved

Functional properties of neurons derived from fetal mouseneurospheres are compatible with those of neuronal precursors in vivo.

Neural stem cells can be propagated in culture as neurospheres, yielding neurons and glial cells upon differentiation. Although the neurosphere model is widely used, the functional properties of the neurosphere-derived neurons have been only partially characterized, and it is unclear whether repeated passaging alters their functional properties. In this study, we analyzed voltage- and transmitter-gated responses in neuron-like cells obtained by differentiating fetal mouse neurospheres at increasing passages in culture. We report that neurons fire overshooting action potentials in response to depolarizing currents up to passage 10 but loose this capability at later passages, as the density of voltage-gated Na+ and K+ currents decreases. In contrast, the immunoreactivity for the neuronal marker β-tubulin remains unaltered up to passage 21, indicating that this marker is not representative of cell function. In almost all neurons, γ-aminobutyric acid (GABA) evoked bicuculline-sensitive whole-cell currents, resulting from the activation of GABAA receptors, which appeared to be excitatory, insofar as the reversal potential of GABA-gated current was about −50 mV. Much smaller currents were elicited by the glutamatergic agonist AMPA, and only occasional responses to glycine were detected. In these functional aspects, neurosphere-derived neurons are similar to immature neurons differentiating in vivo. Therefore, at least for a limited number of passages in vitro, neurospheres provide an adequate model of in vivo neurogenesis

Ligand-independent CXCR2 dimerization

Homo- and hetero-oligomerization have been reported for several G protein-coupled receptors (GPCRs). The CXCR2 is a GPCR that is activated, among the others, by the chemokines CXCL8 (interleukin-8) and CXCL2 (growth-related gene product beta) to induce cell chemotaxis. We have investigated the oligomerization of CXCR2 receptors expressed in human embryonic kidney cells and generated a series of truncated mutants to determine whether they could negatively regulate the wild-type (wt) receptor functions. CXCR2 receptor oligomerization was also studied by coimmunoprecipitation of green fluorescent protein- and V5-tagged CXCR2. Truncated CXCR2 receptors retained their ability to form oligomers only if the region between the amino acids Ala-106 and Lys-163 was present. In contrast, all of the deletion mutants analyzed were able to form heterodimers with the wt CXCR2 receptor, albeit with different efficiency, competing for wt/wt dimer formation. The truncated CXCR2 mutants were not functional and, when coexpressed with wt CXCR2, interfered with receptor functions, impairing cell signaling and chemotaxis. When CXCR2 was expressed with the AMPA-type glutamate receptor GluR1, CXCR2 dimerization was again impaired in a dose-dependent way, and receptor functions were prejudiced. In contrast, CXCR1, a chemokine receptor that shares many similarities with CXCR2, did not dimerize alone or with CXCR2 and when coexpressed with CXCR2 did not impair receptor signaling and chemotaxis. The formation of CXCR2 dimers was also confirmed in cerebellar neuron cells. Taken together, we conclude from these studies that CXCR2 functions as a dimer and that truncated receptors negatively modulate receptor activities competing for the formation of wt/wt dimers