Ester to amide substitution improves selectivity, efficacy and kinetic behavior of a benzodiazepine positive modulator of GABA(A) receptors containing the alpha 5 subunit

We have synthesized and characterized MP-III-022 ((R)-8-ethynyl 6 (2 fluorophenyl)-N,4-dimethyl4H-benzo[f]imidazo[1,5-alpha][1,4]diazepine-3-carboxamide) in vitro and in vivo as a binding- and efficacy selective positive allosteric modulator of GABA(A) receptors containing the alpha 5 subunit (alpha 5GABA(A)Rs). By approximation of the electrophysiological responses which the estimated free rat brain concentrations can induce, we demonstrated that convenient systemic administration of MP-III-022 in the dose range 110 mg/kg may result in a selective potentiation, over a wide range from mild to moderate to strong, of alpha 5 beta gamma 2 GABA(A) receptors. For eliciting a comparable range of potentiation, the widely studied parent ligand SH-053-2'F-R-CH3 containing an ester moiety needs to be administered over a much wider dose range (10-200 mg/kg), but at the price of activating non-alpha 5 GABA(A)Rs as well as the desired alpha 5GABA(A)Rs at the highest dose. At the dose of 10 mg/kg, which elicits a strong positive modulation of alpha 5GABA(A)Rs, MP-III-022 caused mild, but significant muscle relaxation, while at doses 1-10 mg/kg was devoid of ataxia, sedation or an influence on the anxiety level, characteristic for non-selective benzodiazepines. As an amide compound with improved stability and kinetic properties, MP-III-022 may represent an optimized tool to study the influence of alpha 5GABA(A)Rs on the neuronal pathways related to CNS disorders such as schizophrenia, Alzheimer's disease, Down syndrome or autism

LIBER Open Science Roadmap

Embracing Open Science is critical if we are to make science more collaborative, reproducible, transparent and impactful. Open Science undoubtedly has the power to positively influence society, but its implementation is not yet universal. A revolution is required: one which opens up research processes and changes mindsets in favour of a world where policies, tools and infrastructures universally support the growth and sharing of knowledge. Research libraries are well placed to make that revolution happen, and LIBER\u27s Open Science Roadmap outlines the specific actions libraries can take to champion Open Science, both within and beyond their own institutions. As we explain in detail throughout this document, libraries need to advocate for Open Science locally and internationally, to support Open Science through tools and services and to expand the impact of their work through collaboration and partnerships. LIBER has shaped its 2018-2022 Strategy to support and enable Open Science and it is our hope that this Roadmap will help Europe’s research libraries to do the same. This document was written during spring 2018, when the Open Science Policy Platform (OSPP) produced integrated advice for the EC and key stakeholders. People from across the LIBER community translated the OSPP recommendations for libraries and combined them with suggestions drawn from their own expertise and experiences

LIBER Open Science Roadmap

Embracing Open Science is critical if we are to make science more collaborative, reproducible, transparent and impactful. Open Science undoubtedly has the power to positively influence society, but its implementation is not yet universal. A revolution is required: one which opens up research processes and changes mindsets in favour of a world where policies, tools and infrastructures universally support the growth and sharing of knowledge. Research libraries are well placed to make that revolution happen, and LIBER\u27s Open Science Roadmap outlines the specific actions libraries can take to champion Open Science, both within and beyond their own institutions. As we explain in detail throughout this document, libraries need to advocate for Open Science locally and internationally, to support Open Science through tools and services and to expand the impact of their work through collaboration and partnerships. LIBER has shaped its 2018-2022 Strategy to support and enable Open Science and it is our hope that this Roadmap will help Europe’s research libraries to do the same. This document was written during spring 2018, when the Open Science Policy Platform (OSPP) produced integrated advice for the EC and key stakeholders. People from across the LIBER community translated the OSPP recommendations for libraries and combined them with suggestions drawn from their own expertise and experiences

Kif1b is essential for mRNA localization in oligodendrocytes and development of myelinated axons

The kinesin motor protein Kif1b has previously been implicated in the axonal transport of mitochondria and synaptic vesicles1,2. More recently kif1b has been linked with susceptibility to Multiple Sclerosis (MS) 3. Here we show that Kif1b is required for the localization of myelin basic protein mRNA to processes of myelinating oligodendrocytes in zebrafish. We observe the ectopic appearance of myelin-like membrane in kif1b mutants, coincident with the ectopic localization of myelin proteins in kif1b mutant oligodendrocyte cell bodies. These observations suggest the hypothesis that oligodendrocytes localize certain mRNA molecules, namely those encoding small basic proteins such as mbp, to prevent aberrant effects of these proteins elsewhere in the cell. We also find that Kif1b is required for outgrowth of some of the longest axons in the peripheral and central nervous systems. Our data demonstrate new functions of kif1b in vivo and provide insights into its possible roles in Multiple Sclerosis

BMP signaling in astrocytes downregulates EGFR to modulate survival and maturation.

Astrocytes constitute a major cell population in the brain with a myriad of essential functions, yet we know remarkably little about the signaling pathways and mechanisms that direct astrocyte maturation. To explore the signals regulating astrocyte development, we prospectively purified and cultured immature postnatal rodent astrocytes. We identified fibroblast growth factors (FGFs) and bone morphogenetic proteins (BMPs) as robust trophic factors for immature astrocytes. We showed that astrocytes respond directly to BMPs via phosphorylation of the smad1/5/8 pathway. In vitro, BMP signaling promoted immature astrocytes to adopt multiple characteristics of mature astrocytes, including a more process-bearing morphology, aquaporin-4 (AQP4) and S100β immunoreactivity, limited proliferation, and strong downregulation of epidermal growth factor receptor (EGFR). In vivo, activation of the smad1/5/8 pathway in astrocytes was seen during early postnatal development, but inhibition of astrocyte proliferation was not observed. These insights can aid in the further dissection of the mechanisms and pathways controlling astrocyte biology and development

BMP and FGF are robust trophic factors for purified astrocytes.

<p>A–D. Live/Dead viability images at 3DIV of purified astrocyte cultures with control base media (A) containing either HbEGF (B), BMP5 (C), or FGF2 (D). Green cells are alive, red cells are dead. Scale bars are 100 µm. E–I. Quantification of purified astrocyte survival at 3DIV in base media with the addition of various cytokines. 5 ng/ml HbEGF (positive control), as well as 100 ng/ml BMP5, 100 ng/ml IGF2 and 10 ng/ml FGF2 each significantly increase astrocyte viability above control (E). Within the TGF-β superfamily multiple BMP subfamily members also promoted astrocyte survival at 100 ng/ml (F). Additional FGF family members were examined, but only FGF1 also had a modest trophic effect at 10 ng/ml (G). BMP5 survival was dose dependent and trophic effects plateau at 50 ng/ml (H). No additive effects on astrocyte survival above HbEGF alone were observed with any combination of HbEGF, FGF2, and BMP5 (I). N ≥3 for each condition. Significance determined using one-way ANOVA with Dunnett correction. Error bars represent SEM.</p

BMP signaling modulates astrocyte maturation <i>in</i><i>vitro</i>.

<p>A–C. Purified astrocytes were cultured for 3DIV in HbEGF (A), BMP5 (B), or FGF2 and GFAP expression examined by immunostaining. CellMask HCS staining was used to visualize all astrocytes. Images show individual channels from the same field. Scale bars are 100 µm. The percentage of astrocytes expressing GFAP at 3DIV was quantified using ImageJ (C). D–G. Purified astrocytes were cultured for 3DIV in HbEGF or BMP5 and the expression of AQP4 and S100ß examined by immunostaining. All cultures were also stained with GFAP. Scale bars are 50 µm. H–J. The proliferative capabilities of purified astrocytes in HbEGF, BMP5 and FGF2 were quantified <i>in</i><i>vitro</i> over 7 days using clonal analysis. Average clone size was calculated for both individual growth factors (H) and combinations of factors (I). Clonal analysis was also performed on astrocytes pre-treated with BMP5 for 7DIV to examine reversibility (J). K–L. Western blots for EGFR. Protein samples collected from acutely purified astrocytes at different postnatal time points confirm strong developmental downregulation of astrocyte EGFR (K). EGFR is also strongly downregulated in astrocytes cultured with BMP5 for 6DIV, even in the presence of other growth factors (L). Actin bands confirm equal protein loading. N ≥3 for each condition. Significance determined using one-way ANOVA with Tukey correction. Error bars represent SEM.</p