Journal: Molecular Neurodegeneration

Article Title: Activation of Wnt signaling promotes hippocampal neurogenesis in experimental autoimmune encephalomyelitis

doi: 10.1186/s13024-016-0117-0

Figure Lengend Snippet: Hippocampal neurogenesis is increased in the acute phase of EAE. a Induction of pro-neuronal transcription factors in the hippocampus of mice with passive EAE. qPCR analysis of the Wnt -dependent genes ( NeuroD1 ( ND1 ), Prox1 , Lef1) and Dlx2 , a marker of transient amplifying neuroblasts. Histogram represents mean + SEM of fold-changes in gene expression of EAE mice relative to control (PBS) group set as 1. Gapdh was used as an endogenous reference. Day 20 (EAE, n = 4–8; control, n = 4–8); day 50 (EAE, n = 4–10; control, n = 4–11). Two-tailed, unpaired Student’s t -test, * p < 0.05, ** p < 0.01 and *** p < 0.001. b Disease course of C57/B6 EAE mice immunized with encephalitogenic MOG 35–55 . Control group (CFA) were injected with an adjuvant cocktail without antigen. Data are shown as a mean clinical score ± SEM. In acute phase of disease (days 15–20; highlighted by green line) EAE and respective control mice received daily i.p. injections of BrdU (50 mg/kg body weight). After a 2-weeks chase period without administration of BrdU mice were sacrificed for histological assessment (day 30). c Histological analysis of proliferating cells in the DG of EAE mice. Immunostaining for BrdU (green) and Dcx (red) in hippocampal sections. Arrow heads indicate BrdU + /Dcx + co-labeled neuronal progenitors. Nuclei were counterstained with Hoechst (grey). Scale bar, 50 μm. d The frequency of BrdU label-retaining cells in the DG is increased in mice with EAE (day 30; n = 3 mice; 18–28 sections per mouse) as compared to control mice ( n = 3 mice; 17–28 sections per mouse). Data is shown as mean + SEM of positive cells per mm of the DG. Two-tailed, unpaired Student’s t -test,* p < 0.05 and ** p < 0.01

Article Snippet: Sections were incubated with primary rat anti-BrdU antibody (1:400; Upstate ) and goat anti-Dcx antibody (1:500; Santa Cruz Biotechnology) overnight at 4 °C, followed by incubation with appropriate Cy2/Cy3-conjugated secondary antibodies (Millipore) for 1 h at RT.

Techniques: Marker, Expressing, Two Tailed Test, Injection, Immunostaining, Labeling

Journal: Molecular Psychiatry

Article Title: Revisiting adult neurogenesis and the role of erythropoietin for neuronal and oligodendroglial differentiation in the hippocampus

doi: 10.1038/mp.2015.212

Figure Lengend Snippet: Effects of erythropoietin (EPO) on neuronal differentiation and maturation, determined by mean active area of synapses in E17-HCC. Neurons (d8) were quadruply stained for differentiation markers Dcx and MAP2 and activity markers Synaptotagmin I (SytI) and Synapsin I (SynI) ( n =8). Staining was analyzed by confocal microscopy. ( a ) The protocol includes three sequential scans with fixed emission windows (orange) with different excitation wave length. Excitation (dashed line) and emission (solid line) spectra are shown for each fluorescent dye, Alexa Fluor 488 (MAP2, green), Alexa Fluor 546 (SynI, blue), Alexa Fluor 594 (SytI, red) and Alexa Fluor 633 (Dcx, black). 4,6-Diamidino-2-phenylindole (DAPI) fluorescence was determined using an additional excitation at 406 nm. ( b ) Unmixed confocal picture showing differentiation markers Dcx (red), MAP2 (green) and DAPI (blue). ( c ) Quantification of the integrated density of Dcx and MAP2 presented as ratio ( n =8 per group, paired one-tailed t -test). ( d ) Unmixed confocal picture showing SytI (green) and SynI (red). ( e ) Higher magnification of the square mark of picture d , showing single stained dots for SytI (green, arrowhead), single stained dots for SynI (red, star) and colocalized dots (yellow, arrow). ( f ) Masked images for analyzing the number and area of SytI (green, arrowhead), SynI (red, star) and colocalized (white, arrow) dots. ( g ) Quantification of the mean overlapping (colocalized) area as mean active area (white spots in f , n =8 per group, paired two-tailed t -test). All n-numbers given are derived from biological replicates, that is, independent cell preparations. All bar graphs are shown as mean±s.e.m.; * P <0.05.

Article Snippet: Sections were then washed and incubated with the direct-labeled mouse anti-BrdU Alexa Fluor 594 (1:500, Invitrogen) in 3% NHS, 0.5% Triton-X in PBS for 48 h at 4 °C.

Techniques: Staining, Activity Assay, Confocal Microscopy, Fluorescence, One-tailed Test, Two Tailed Test, Derivative Assay

Journal: Parasites & Vectors

Article Title: Differential alterations in the small intestine epithelial cell turnover during acute and chronic infection with Echinostoma caproni (Trematoda)

doi: 10.1186/s13071-015-0948-5

Figure Lengend Snippet: a Immunohistochemistry anti-BrdU on small intestine tissue sections from control and infected mice and rats 1 h after BrdU intraperitoneal administration (wpi: week post infection). Cell nuclei were stained with DAPI (blue) and BrdU-positive nuclei are pink. Scale bar: 10 μm. b Number of cells per crypt of Lieberkühn and c percentage of BrdU-labeled cells per crypt 1 h after BrdU administration in mice (white bars) and rats (black bars) at different wpi. Results are presented as mean ± standard deviation. Asterisks indicate significant differences with respect to uninfected controls (0 wpi) for each host species (* p < 0.001). Horizontal bars indicate differences between infected animals at different wpi (a: p < 0.05)

Article Snippet: Immunostaining of BrdU was performed on 4 μm paraffin sections, using a monoclonal anti-BrdU antibody (mouse anti-BrdU MoBU-1 clone, Life Technologies™) labeled with Alexa® Fluor 555dye (excitation/emission maxima: 555/580 nm).

Techniques: Immunohistochemistry, Infection, Staining, Labeling, Standard Deviation