Journal: Molecular Psychiatry

Article Title: The bacterial peptidoglycan-sensing molecule Pglyrp2 modulates brain development and behavior

doi: 10.1038/mp.2016.182

Figure Lengend Snippet: Expression and cellular localization of PGRPs in the developing brain. Changes in striatal Pglyrp2 ( a ) and Pglyrp3 ( b ) protein levels during postnatal development. Protein levels were normalized to the Hsp90 protein levels and expressed relative to the adult group (that is, P60; n =4 per group). All data ( a and b ) are presented as means (±s.e.m.). * P <0.05, ** P <0.01 and *** P <0.001 when compared with their respective adult groups (that is, P60). Distribution of Pglyrp2 in the cerebellum ( c ), frontal cortex ( d – f ) and hippocampus ( g – i ) at P3. Double immunofluorescence staining shows staining of Pglyrp2 (green) combined with either NeuN (a neuronal marker; red), Iba1 (a microglial marker; red), GFAP (an astrocyte marker; red) or calbindin (a marker for purkinje cells; red). The merged images show the colocalization of Pglyrp2 (green) and NeuN (red) in the frontal cortex ( d ) and hippocampus ( g ) and with calbindin (red) in the cerebellum ( c ). Note that Pglyrp2 is moderately colocalization with astrocytes ( f and i ), and with microglia to a lesser extend ( e and h ). Arrows indicate Pglyrp2-positive Purkinje cells (c), Pglyrp2-positive microglia ( e and h ) and Pglyrp2-positive astrocytes ( f and i ). The scale bars represent 50 μm. Hsp, heat shock protein; P, postnatal day; PGRP, peptidoglycan-recognition protein.

Article Snippet: For double labelling, the following antisera were used: mouse monoclonal antibody against NeuN to label mature neurons (cat no. MAB377, 1:250; Merck Millipore, Billerica, MA, USA), mouse monoclonal antibody against Calbindin to label Purkinje cells in the cerebellum (cat no. AB82812, 1:500; Abcam; Cambridge, UK), goat polyclonal antibody against GFAP to label astrocytes (cat no. AB53554, 1:500; Abcam), and goat polyclonal antibody against Iba1 to label microglial cells (cat no. AB5076, 1:800; Abcam), and were detected using either Donkey Anti-Mouse Cy5 antiserum or Donkey Anti-Goat Cy5 antiserum (1:250; Jackson ImmunoResearch Europe, Suffolk, UK).

Techniques: Expressing, Double Immunofluorescence Staining, Staining, Marker

Journal: PLoS ONE

Article Title: Short-term succinic acid treatment mitigates cerebellar mitochondrial OXPHOS dysfunction, neurodegeneration and ataxia in a Purkinje-specific spinocerebellar ataxia type 1 (SCA1) mouse model

doi: 10.1371/journal.pone.0188425

Figure Lengend Snippet: (A) Representative calbindin-stained cerebellar primary fissure images from untreated wild type, untreated B05 and treated B05 mice. (B) Molecular layer thickness, a measure of calbindin-positive Purkinje dendritic length, in treated and untreated wild type and B05 cerebellum. (C) Representative ATXN1-stained cerebellar primary fissure images from untreated control transgenic (A02), untreated B05 and treated B05 mice. (D) Purkinje nuclei counts in treated and untreated A02 and B05 cerebellum. Error bars represent means ± SEM. Two-way ANOVA,* P < 0.05, ** P < 0.01.

Article Snippet: Following an established protocol [ ], epitopes were unmasked by boiling three times for 15 seconds each in 0.01 M urea, blocked for 1 hour in 2% normal donkey serum and 0.3% Triton X-100 in 1X PBS, and incubated for 48 hours at 4°C in blocking solution containing goat calbindin antibody (SC-7691, Santa Cruz) at 1:500.

Techniques: Staining, Control, Transgenic Assay