Ethanol withdrawal (EW) decreases GluN2A mRNA levels.

<p>Levels of mRNA transcripts (measured by quantitative PCR) for the N-methyl-D-aspartate (NMDA) receptor subunits, CB1 receptors, and FAAH enzyme in cultured cortical neurons in control conditions (neurons not exposed to ethanol) or during ethanol withdrawal (EW group). Values are normalized to β-actin gene and expressed as percentages over control data (n = 3–4 samples/group; means ± SEM). Data were assessed by the Student's t-test (* <i>P</i><0.05 vs. control; a, p = 0.097 vs. control).</p

The acute manipulation of the endocannabinoid system influences neuronal viability during ethanol withdrawal.

<p>Effects of the acute activation or blockade of the endocannabinoid signaling on neuronal viability in control (neurons not exposed to ethanol) or ethanol–withdrawn neurons (EW). (A) Representative scheme of the experimental protocol. (B) The administration of the non-specific cannabinoid agonist HU-210 (1 µM) during ethanol withdrawal decreases NMDA-induced neuronal death in ethanol-withdrawn neurons. Interestingly, the neuroprotective effect of HU-210 is specific of alcohol withdrawal situation, since it has no effect on control neurons. (C) The acute administration of the CB1 antagonist rimonabant (1 µM) increases NMDA-induced neuronal death in alcohol-withdrawn neurons. Again, this effect is specific of the withdrawal situation and rimonabant, when administered acutely, has no effect on control condition. Values are normalized to NMDA and are means ± SEM (n = 14–18 wells/condition; N = 4 plates). Data were analyzed by the ANOVA test followed by the Fisher's PLSD test (* <i>P</i><0.05).</p

Sequences of primers used for quantitative PCR.

<p>Sequences of primers used for quantitative PCR.</p

Ethanol withdrawal increases by 40% the sensitivity of neurons to excitotoxic injuries induced by NMDA (10 µM, 24 h) in cultured cortical neurons.

<p>(A) Phase-contrast photomicrographs show representative fields in the indicated conditions (Control, neurons not exposed to ethanol; EW, ethanol-withdrawn neurons). (B) Quantification of neuronal death. Values are means ± SEM (n = 17–20 wells/condition; N = 5 plates). Data were assessed by ANOVA test followed by Fisher's PLSD test (* <i>P</i><0.05; *** <i>P</i><0.001).</p

The stimulation of the endocannabinoid system reduces NMDA-stimulated calcium influx in ethanol-withdrawn neurons.

<p>(A, B, C) Representative recordings of single-cell calcium videomicroscopy for each experiment. A 30 sec exposure to NMDA (50 µM) produced a rapid calcium influx, which recovered over the following 2 min (<i>before</i>). (E) <i>After</i> the incubation (10 min) with the non-specific CB1 agonist HU-210 (1 µM), Ca²⁺ influx (induced by NMDA application) is significantly decreased in comparison to the stimulation <i>before</i>. Incubation (10 min) with DMSO (D) or rimonabant (1 µM) (F) does not modify NMDA-stimulated calcium influx. Values are expressed as % vs. <i>before</i> NMDA-stimulation ± SEM (N = 3–4; n (cell number) >60 cells). Data were analyzed by the t Student's test. (***<i>P</i><0.001).</p

The chronic manipulation of the endocannabinoid system influences neuronal viability during ethanol withdrawal.

<p>Effects of the chronic activation or blockade of the endocannabinoid signaling on neuronal viability in control (neurons not exposed to ethanol) or ethanol–withdrawn neurons (EW). (A) Representative scheme of the experimental protocol. (B) The chronic administration of the non-specific cannabinoid agonist HU-210 (1 µM) decreases NMDA-induced neuronal death during ethanol withdrawal. The neuroprotective effect of HU-210 is specific of ethanol withdrawal situation, since it has no effect on the control condition. (C) The chronic administration of the CB1 antagonist rimonabant (1 µM) increases neuronal death. Moreover, the long-term administration of rimonabant tends to be neurotoxic not only for ethanol-withdrawn neurons but also for control neurons. Values are normalized to NMDA and are means ± SEM (n = 6–16 wells/condition; N = 3–4 plates). Data were analyzed by the ANOVA test followed by the Fisher's PLSD test. (*<i>P</i><0.05; ** <i>P</i><0.01; a, p = 0.0551).</p

Therapeutic Benefits from Nanoparticles: The Potential Significance of Nanoscience in Diseases with Compromise to the Blood Brain Barrier

Therapeutic Benefits from Nanoparticles: The Potential Significance of Nanoscience in Diseases with Compromise to the Blood Brain Barrie

Additional file 3: Figure S2. of Tissue-type plasminogen activator exerts EGF-like chemokinetic effects on oligodendrocytes in white matter (re)myelination

EGFR is expressed in early oligodendrocyte precursors. Photomicrographs from embryonic spinal cord (ventral ventricular zone, E13) and perilesional adult corpus callosum (3dpi) WT mice tissue sections show representative confocal images of EGFR (green), Sox2 or PDGFRα (red) and DAPI (blue) immunoreactivities. Asterisks indicate PDGFRα+/EGFR− cells and arrowheads indicate PDGFRα+/EGFR+ cells (Representative images from n = 3). Scale bars: 10 μm. (TIF 585 kb

Additional file 1: Table S1. of Tissue-type plasminogen activator exerts EGF-like chemokinetic effects on oligodendrocytes in white matter (re)myelination

Antibodies used in the study. Each antibody used in this study is listed together with its supplier, species, type (monoclonal/polyclonal), dilution and reference. (DOCX 16 kb

Additional file 4: Figure S3. of Tissue-type plasminogen activator exerts EGF-like chemokinetic effects on oligodendrocytes in white matter (re)myelination

tPA is not detected in PDFGR-β + pericytes. (A) Photomicrographs from adult WT mice tissue sections show representative confocal images of PDGFR-β (green), tPA (red) and DAPI (blue) immunoreactivities in the lesion 3 days after injection. (Representative images from n = 3). Inlets show side view reconstructions of the z-stack. (B) Representative fluorescent intensity/distance graph measured from confocal image in (A) showing that tPA (Red) is not found in colocalization with PDGFR-β staining. (TIF 328 kb