Structure and function of an RNase H domain at the heart of the spliceosome.

Precursor-messenger RNA (pre-mRNA) splicing encompasses two sequential transesterification reactions in distinct active sites of the spliceosome that are transiently established by the interplay of small nuclear (sn) RNAs and spliceosomal proteins. Protein Prp8 is an active site component but the molecular mechanisms, by which it might facilitate splicing catalysis, are unknown. We have determined crystal structures of corresponding portions of yeast and human Prp8 that interact with functional regions of the pre-mRNA, revealing a phylogenetically conserved RNase H fold, augmented by Prp8-specific elements. Comparisons to RNase H–substrate complexes suggested how an RNA encompassing a 5′-splice site (SS) could bind relative to Prp8 residues, which on mutation, suppress splice defects in pre-mRNAs and snRNAs. A truncated RNase H-like active centre lies next to a known contact region of the 5′SS and directed mutagenesis confirmed that this centre is a functional hotspot. These data suggest that Prp8 employs an RNase H domain to help assemble and stabilize the spliceosomal catalytic core, coordinate the activities of other splicing factors and possibly participate in chemical catalysis of splicing

DANSSino: a pilot version of the DANSS neutrino detector

DANSSino is a reduced pilot version of a solid-state detector of reactor antineutrinos (to be created within the DANSS project and installed under the industrial 3 GW(th) reactor of the Kalinin Nuclear Power Plant -- KNPP). Numerous tests performed at a distance of 11 m from the reactor core demonstrate operability of the chosen design and reveal the main sources of the background. In spite of its small size (20x20x100 ccm), the pilot detector turned out to be quite sensitive to reactor antineutrinos, detecting about 70 IBD events per day with the signal-to-background ratio about unity.Comment: 16 pages, 11 figures, 3 tables. arXiv admin note: substantial text overlap with arXiv:1304.369

Developmental changes in electrophysiological properties and a transition from electrical to chemical coupling between excitatory layer 4 neurons in the rat barrel cortex

© 2016 Valiullina, Akhmetshina, Nasretdinov, Mukhtarov, Valeeva, Khazipov and Rozov.During development, sensory systems switch from an immature to an adult mode of function along with the emergence of the active cortical states. Here, we used patch-clamp recordings from neocortical slices in vitro to characterize the developmental changes in the basic electrophysiological properties of excitatory L4 neurons and their connectivity before and after the developmental switch, which occurs in the rat barrel cortex in vivo at postnatal day P8. Prior to the switch, L4 neurons had higher resting membrane potentials, higher input resistance, lower membrane capacity, as well as action potentials (APs) with smaller amplitudes, longer durations and higher AP thresholds compared to the neurons after the switch. A sustained firing pattern also emerged around the switch. Dual patch-clamp recordings from L4 neurons revealed that recurrent connections between L4 excitatory cells do not exist before and develop rapidly across the switch. In contrast, electrical coupling between these neurons waned around the switch. We suggest that maturation of electrophysiological features, particularly acquisition of a sustained firing pattern, and a transition from the immature electrical to mature chemical synaptic coupling between excitatory L4 neurons, contributes to the developmental switch in the cortical mode of function

Genetically altered AMPA-type glutamate receptor kinetics in interneurons disrupt long-range synchrony of gamma oscillation

Gamma oscillations synchronized between distant neuronal populations may be critical for binding together brain regions devoted to common processing tasks. Network modeling predicts that such synchrony depends in part on the fast time course of excitatory postsynaptic potentials (EPSPs) in interneurons, and that even moderate slowing of this time course will disrupt synchrony. We generated mice with slowed interneuron EPSPs by gene targeting, in which the gene encoding the 67-kDa form of glutamic acid decarboxylase (GAD67) was altered to drive expression of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate receptor subunit GluR-B. GluR-B is a determinant of the relatively slow EPSPs in excitatory neurons and is normally expressed at low levels in γ-aminobutyric acid (GABA)ergic interneurons, but at high levels in the GAD-GluR-B mice. In both wild-type and GAD-GluR-B mice, tetanic stimuli evoked gamma oscillations that were indistinguishable in local field potential recordings. Remarkably, however, oscillation synchrony between spatially separated sites was severely disrupted in the mutant, in association with changes in interneuron firing patterns. The congruence between mouse and model suggests that the rapid time course of AMPA receptor-mediated EPSPs in interneurons might serve to allow gamma oscillations to synchronize over distance

Causal interrogation of neuronal networks and behavior through virally transduced ivermectin receptors

© 2016 Obenhaus, Rozov, Bertocchi, Tang, Kirsch, Betz and Sprengel.The causal interrogation of neuronal networks involved in specific behaviors requires the spatially and temporally controlled modulation of neuronal activity. For long-term manipulation of neuronal activity, chemogenetic tools provide a reasonable alternative to short-term optogenetic approaches. Here we show that virus mediated gene transfer of the ivermectin (IVM) activated glycine receptor mutant GlyRα1AG can be used for the selective and reversible silencing of specific neuronal networks in mice. In the striatum, dorsal hippocampus, and olfactory bulb, GlyRα1AG promoted IVM dependent effects in representative behavioral assays. Moreover, GlyRα1AG mediated silencing had a strong and reversible impact on neuronal ensemble activity and c-Fos activation in the olfactory bulb. Together our results demonstrate that long-term, reversible and re- inducible neuronal silencing via GlyRα1AG is a promising tool for the interrogation of network mechanisms underlying the control of behavior and memory formation

Final results of the EDELWEISS-II WIMP search using a 4-kg array of cryogenic germanium detectors with interleaved electrodes

The EDELWEISS-II collaboration has completed a direct search for WIMP dark matter with an array of ten 400-g cryogenic germanium detectors in operation at the Laboratoire Souterrain de Modane. The combined use of thermal phonon sensors and charge collection electrodes with an interleaved geometry enables the efficient rejection of gamma-induced radioactivity as well as near-surface interactions. A total effective exposure of 384 kg.d has been achieved, mostly coming from fourteen months of continuous operation. Five nuclear recoil candidates are observed above 20 keV, while the estimated background is 3.0 events. The result is interpreted in terms of limits on the cross-section of spin-independent interactions of WIMPs and nucleons. A cross-section of 4.4x10^-8 pb is excluded at 90%CL for a WIMP mass of 85 GeV. New constraints are also set on models where the WIMP-nucleon scattering is inelastic.Comment: 23 pages, 5 figures; matches published versio

A search for low-mass WIMPs with EDELWEISS-II heat-and-ionization detectors

We report on a search for low-energy (E < 20 keV) WIMP-induced nuclear recoils using data collected in 2009 - 2010 by EDELWEISS from four germanium detectors equipped with thermal sensors and an electrode design (ID) which allows to efficiently reject several sources of background. The data indicate no evidence for an exponential distribution of low-energy nuclear recoils that could be attributed to WIMP elastic scattering after an exposure of 113 kg.d. For WIMPs of mass 10 GeV, the observation of one event in the WIMP search region results in a 90% CL limit of 1.0x10^-5 pb on the spin-independent WIMP-nucleon scattering cross-section, which constrains the parameter space associated with the findings reported by the CoGeNT, DAMA and CRESST experiments.Comment: PRD rapid communication accepte

Muon-induced background in the EDELWEISS dark matter search

A dedicated analysis of the muon-induced background in the EDELWEISS dark matter search has been performed on a data set acquired in 2009 and 2010. The total muon flux underground in the Laboratoire Souterrain de Modane (LSM) was measured to be $\Phi_{\mu}=(5.4\pm 0.2 ^{+0.5}_{-0.9})$\,muons/m$^2$/d. The modular design of the muon-veto system allows the reconstruction of the muon trajectory and hence the determination of the angular dependent muon flux in LSM. The results are in good agreement with both MC simulations and earlier measurements. Synchronization of the muon-veto system with the phonon and ionization signals of the Ge detector array allowed identification of muon-induced events. Rates for all muon-induced events $\Gamma^{\mu}=(0.172 \pm 0.012)\, \rm{evts}/(\rm{kg \cdot d})$ and of WIMP-like events $\Gamma^{\mu-n} = 0.008^{+0.005}_{-0.004}\, \rm{evts}/(\rm{kg \cdot d})$ were extracted. After vetoing, the remaining rate of accepted muon-induced neutrons in the EDELWEISS-II dark matter search was determined to be $\Gamma^{\mu-n}_{\rm irred} < 6\cdot 10^{-4} \, \rm{evts}/(\rm{kg \cdot d})$ at 90%\,C.L. Based on these results, the muon-induced background expectation for an anticipated exposure of 3000\,\kgd\ for EDELWEISS-3 is $N^{\mu-n}_{3000 kg\cdot d} < 0.6$ events.Comment: 21 pages, 16 figures, Accepted for publication in Astropart. Phy

The Large Enriched Germanium Experiment for Neutrinoless Double Beta Decay (LEGEND)

The observation of neutrinoless double-beta decay (0${\nu}{\beta}{\beta}$) would show that lepton number is violated, reveal that neutrinos are Majorana particles, and provide information on neutrino mass. A discovery-capable experiment covering the inverted ordering region, with effective Majorana neutrino masses of 15 - 50 meV, will require a tonne-scale experiment with excellent energy resolution and extremely low backgrounds, at the level of $\sim$0.1 count /(FWHM$\cdot$t$\cdot$yr) in the region of the signal. The current generation $^{76}$Ge experiments GERDA and the MAJORANA DEMONSTRATOR utilizing high purity Germanium detectors with an intrinsic energy resolution of 0.12%, have achieved the lowest backgrounds by over an order of magnitude in the 0${\nu}{\beta}{\beta}$ signal region of all 0${\nu}{\beta}{\beta}$ experiments. Building on this success, the LEGEND collaboration has been formed to pursue a tonne-scale $^{76}$Ge experiment. The collaboration aims to develop a phased 0${\nu}{\beta}{\beta}$ experimental program with discovery potential at a half-life approaching or at $10^{28}$ years, using existing resources as appropriate to expedite physics results.Comment: Proceedings of the MEDEX'17 meeting (Prague, May 29 - June 2, 2017