Journal: Mediators of Inflammation

Article Title: The Astrocytic S100B Protein with Its Receptor RAGE Is Aberrantly Expressed in SOD1 G93A Models, and Its Inhibition Decreases the Expression of Proinflammatory Genes

doi: 10.1155/2017/1626204

Figure Lengend Snippet: S100B increase in GFAP-positive cells in the lumbar spinal cord from SOD1 G93A rats. Double immunofluorescence labelling was performed with anti-S100B (green) and anti-GFAP (red) in the lumbar spinal cord from wild-type (WT), presymptomatic (ps-SOD1 G93A ), and end-stage (es-SOD1 G93A ) rats. The images were acquired from the grey matter (GM) and from the white matter (WM) at two magnifications (scale bars: 50 μ m left columns and 20 μ m right column). Merged panels also show To-Pro-3 nuclear staining (TP3, blue). In WM from WT rats, insets show a double immunolabelling with anti-S100B (green) and anti-CNPase (red) and merged inset also with To-Pro-3 staining (scale bar: 10 μ m).

Article Snippet: The following primary antibodies were used for immunofluorescence: anti-rabbit S100B (1 : 7500, Novus Biological), anti-mouse S100B (1 : 1000, Sigma Aldrich), anti-mouse RAGE (1 : 200, Millipore), anti-rabbit GFAP (1 : 1000, Dako), anti-mouse GFAP (1 : 1000, Novus Biologicals), anti-mouse NeuN (1 : 500, Millipore), anti-rabbit Iba1 (1 : 200, Wako), anti-mouse CNPase (1 : 500, Novus Biologicals), and anti-rabbit ChAT (1 : 200, Millipore).

Techniques: Immunofluorescence, Staining

Journal: Neural Regeneration Research

Article Title: Trophic factors are essential for the survival of grafted oligodendrocyte progenitors and for neuroprotection after perinatal excitotoxicity

doi: 10.4103/1673-5374.266066

Figure Lengend Snippet: Representative views of the CC 35 days after treatment. In the presence of NMDA (N) alone there was considerable nestin-expressing cell loss and extreme reaction of blood vessels that auto-fluoresced. Very few cells, most likely pericytes, displayed intense GFP-nestin fluorescence. (B) View of the same field where blood vessels were conspicuously auto-fluorescent, but no cells in the brain parenchyma displayed CNPase labeling. (C) The merged image allows for the visualization of auto-fluorescent blood vessels as well as very few nestin-positive/CNPase-negative cells (open arrows). (D) In animals that received N + TSC1 the CC displayed very few GFP-expressing cells besides two or three of them (open arrows). (E) Blood vessels were not auto-fluorescent and the tissue presented its cytoarchitecture undisturbed, yet only few single cells expressed CNPase in their soma (arrows). Most tissue in the CC showed a diffuse-positive CNPase label. (F). The merged image allows for the visualization of auto-fluorescent blood vessels as well as the nestin-positive/CNPase-negative cells (arrows). (G) When mice received NMDA + CG4 grafts, very few nestin expressing cells were seen in the SVZ and virtually none in the brain parenchyma. (H) The tissue displayed no CNPase immunoreactivity, yet there were significantly less blood vessels revealed by auto-fluorescence. (I) The merged image shows these cells (FB shown in magenta pseudo-color). (J) Not many FB-labeled CG4-OLPs were found in these mice but those present were either small round cells (arrowheads) or flat and more fibrous, reactive-like ones. (K) When mice received NMDA (N) + TSC1 + CG4-OLPs, the parenchyma was better preserved and two subpopulations of nestin expressing cells were present. Flat fibrous reactive-like (arrows) and single small round nestin-GFP (arrowheads). (L) The combined treatment of OLPs and TSC1 showed immunoreactivity for CNPase and the cytoarchitecture of the parenchyma was very well preserved. (M) Numerous grafted FB-labeled CG4 cells survived. From the small round cell subpopulation some expressed CNPase (an OL marker and the FB-label seen in pseudo-color magenta), these cells were nestin negative. (N) Numerous grafted OLPs, the cytoplasmic distribution of CNPase is shown by arrows. Rows of grafted cells are shown by arrowheads. Insets show higher magnification of bipolar or multipolar grafted cells and their cell processes in more detail. (O) Diagramatic representation of a sagittal view of mouse brain indicating a single site of injection and the regions from which pictures were taken. CC: Corpus callosum; CG4: central glial 4 cells; CNPase: cyclic nucleotide 3′-phosphohydrolase; FB: Fast Blue; GFP: green fluorescent protein; NMDA: N-methyl-D-aspartate; OLPs: oligodendrocyte progenitors; SVZ: subventricular zone; TSC1: the combination of transferrin and insulin growth factor 1.

Article Snippet: HSP90-alpha mouse monoclonal antibody (MA5-32057, 1:200 dilution, ThermoFisher Scientific, Waltham, MA USA); CNPase rabbit polyclonal antibody (100-401-D21, 1:50 dilution, Rockland Immunochemicals Inc., Limerick, PA, USA).

Techniques: Expressing, Fluorescence, Labeling, Marker, Injection

Journal: Neural Regeneration Research

Article Title: Trophic factors are essential for the survival of grafted oligodendrocyte progenitors and for neuroprotection after perinatal excitotoxicity

doi: 10.4103/1673-5374.266066

Figure Lengend Snippet: Representative views of the CC and striatum 35 days after treatment. (A) In the presence of NMDA alone there was considerable nestin-expression loss in the brain parenchyma, just a few nestin-expressing cells were present in the SVZ. Nonetheless, pericytes were nestin-GFP labeled. The blood vessels were auto-fluorescent and the tissue presented a spongy aspect in the striatum, the white outline shows tissue loss. (B) CNPase expression was absent from the white matter regions. (C) The merged image illustrates better the presence of few nestin-expressing cells in the apical portion of the SVZ and the auto-fluorescence of capillaries. The absence of CNPase is clearly appreciated. (D) The injection of NMDA followed by TSC1 3 days after the excitotoxic insult resulted in auto-fluorescence quenching, loss of nestin-expressing cells and some tissue loss in this region. (E) Two kinds of cells expressed CNPase small cells organized as rows in the CC and large flat and fibrous-like cells. (F) The merged image confirms the presence of small nestin-positive cells and some tissue loss has been outlined. (G) When central glial 4 cells-oligodendrocyte progenitors (CG4-OLPs) were injected with NMDA nestin expression was absent in the striatum, and just a few blood vessels were auto-fluorescent. (H) The tissue appears spongy and there is not CNPase expression. (I) The merged picture shows the presence of a few grafted cells and autofluorescent blood vesels. (J) A few FB-labeled grafted cells seen in pseudo-color magenta (arrows) distributed in just one region of the tissue. (K) View of the effects of NMDA with the combinatorial treatment of TSC1 + CG4-OLP grafts where we can appreciate nestin-GFP-expressing cells in the SVZ and the wall of the ventricle. (L) Some small cells expressed CNPase (arrows). (M) The merged picture shows grafted cells intermingled with nestin-labeled host cells (short arrows) or adjacent to the nestin-expressing host cells. This image allows for the visualization of FB + cells (seen in magenta) arranged in straight and long rows starting from the SVZ towards the striatum brain parenchyma (long arrows with short heads) point to some of these cell rows. At the level of the SVZ FB-grafted cells and nestin-GFP host cells appear to overlap showing spots of yellow fluorescence (medium size arrows). Inset shows in detail FB-labeled cells and host nestin-expressing cells. (N) The striatum was populated by CG4-OLPs FB-labeled (magenta, short headed long arrows). small arrow points to a grafted cell in the SVZ. (O) Diagramatic representation of a sagittal view of mouse brain indicating a single site of injection and the regions from which pictures were taken. CC: Corpus callosum; CG4: central glial 4 cells; CNPase: cyclic nucleotide 3′-phosphohydrolase; FB: Fast Blue; GFP: green fluorescent protein; NMDA: N-methyl-D-aspartate; OLPs: oligodendrocyte progenitors; SVZ: subventricular zone; TSC1: the combination of transferrin and insulin growth factor 1.

Article Snippet: HSP90-alpha mouse monoclonal antibody (MA5-32057, 1:200 dilution, ThermoFisher Scientific, Waltham, MA USA); CNPase rabbit polyclonal antibody (100-401-D21, 1:50 dilution, Rockland Immunochemicals Inc., Limerick, PA, USA).

Techniques: Expressing, Labeling, Fluorescence, Injection