Journal: Frontiers in Cellular Neuroscience

Article Title: Chronic Stress Modulates Interneuronal Plasticity: Effects on PSA-NCAM and Perineuronal Nets in Cortical and Extracortical Regions

doi: 10.3389/fncel.2019.00197

Figure Lengend Snippet: List of primary and secondary antibodies used in the study.

Article Snippet: WFA lectin biotin-conjugated , 1:200 , Sigma.

Techniques: Avidin-Biotin Assay

Journal: bioRxiv

Article Title: Hyperactive MEK1 signaling in cortical GABAergic neurons causes embryonic parvalbumin-neuron death and defects in behavioral inhibition

doi: 10.1101/748087

Figure Lengend Snippet: (A-F) Representative high-resolution confocal Airyscan images of triple immunolabeled cortical sections for Ai3/EYFP, VGAT, and NEUN. Excitatory neuron perisomatic domains were outlined and quantification of VGAT-labeled pixels revealed that mutants (D-F) have a significant reduction in the amount of perisomatic VGAT-labeling (Scale bar = 3 µm) in comparison to controls (A-C) (quantification in M ; n = 48 control, 53 mutant neurons; mean ± SEM, * = p < 0.05). (G-L) P60 representative coronal sections of Slc32A1:Cre Ai3 (G-I) and caMek1 Slc32A1:Cre Ai3 (J-L) cortices immunolabeled for GFP, WFA, and PV. The WFA channel was imaged using the same acquisition settings for all samples. A significant increase in WFA-labeled area per neuron was detected in mutant cortices when compared to controls (quantification in N ; n = 63 control, 54 mutant neurons, mean ± SEM, * = p < 0.001). Analysis of WFA-labeling intensity yielded a significant increase in integrated density in mutant CINs (quantification in O ; n = 63 control, 54 mutant neurons, mean ± SEM, * = p < 0.001). (Scale bar = 100 µm).

Article Snippet: The primary antibodies used in these experiments were: goat anti-parvalbumin (Swant, 1:1000), rabbit anti-somatostatin (Peninsula, 1:1000), biotin-conjugated WFA (Vector, 60ug/mL), chicken anti-GFP (Aves, 1:1000), chicken anti-RFP (Rockland, 1:1000), rabbit anti-P-ERK (Cell Signaling, 1:1000), rabbit anti-MEK1 (Abcam, 1:1000), rabbit anti-ERK2 (Abcam, 1:1000), rabbit anti P-ERK1/2 (Cell Signaling, 1:1000), mouse anti-NEUN (Millipore 1:1000), rabbit anti-cleaved caspase 3 (Cell Signaling, 1:1000), rabbit anti-GFAP (Abcam 1:1000), rabbit anti-VGAT (Synaptic Systems, 1:1000), and mouse anti-8-oxo-DG (R&D Systems, 1:1000) ( ).

Techniques: Immunolabeling, Labeling, Mutagenesis

Journal: Frontiers in Molecular Neuroscience

Article Title: The Perineuronal ‘Safety’ Net? Perineuronal Net Abnormalities in Neurological Disorders

doi: 10.3389/fnmol.2018.00270

Figure Lengend Snippet: Analysis of PV cells enwrapped with PNNs across sensory cortices.

Article Snippet: , C57BL/6J , PV: (1) monoclonal mouse α-PV (SWANT, 1:500); (2) polyclonal guinea pig α-PV PNN: biotin-conjugated WFA (Sigma, 1:200) , All layers P18: ∼55% P35: ∼55%.

Techniques: Plasmid Preparation

Journal: Journal of Neurophysiology

Article Title: Diaphragm electromyographic activity following unilateral midcervical contusion injury in rats

doi: 10.1152/jn.00727.2016

Figure Lengend Snippet: Extent of perineuronal net formation around injury epicenter at 7 days following a unilateral C4 contusion injury. A: representative 20 μm-thick transverse sections from an individual unilateral C4 contusion animal displaying Wisteria floribunda agglutinin (WFA) immunoreactivity (grayscale; black reflects greater immunoreactivity) in 5 sections (∼200 μm apart), spanning 0.4 mm rostral to 0.4 mm caudal to the injury epicenter. An outline of the spinal cord section and contralateral gray matter is presented for clarity. B: summary of integrated WFA immunoreactivity (fluorescence intensity), 2.4 mm around the injury epicenter in laminectomy (n = 4) and contusion (n = 4) groups. A significant increase in perineuronal net formation was evident at a distance 0.4 mm rostral to 0.6 mm caudal to the injury site (*P < 0.001). Original scale bar, 1 mm.

Article Snippet: One series of sections was used to label the perineuronal net using biotin-conjugated Wisteria floribunda agglutinin (WFA; Vector Laboratories, Burlingame, CA) by incubating serially in unlabeled streptavidin and biotin, biotin-WFA (1:200), and streptavidin Alexa Fluor 568 (1:50; Thermo Fisher Scientific).

Techniques: Fluorescence

Journal: Frontiers in Molecular Neuroscience

Article Title: The Perineuronal ‘Safety’ Net? Perineuronal Net Abnormalities in Neurological Disorders

doi: 10.3389/fnmol.2018.00270

Figure Lengend Snippet: Analysis of PV cells enwrapped with PNNs across sensory cortices.

Article Snippet: , C57BL/6J , PV: monoclonal mouse α-PV (Sigma, 1:1000) PNN: biotin-conjugated WFA (Vector, 1:200) , All layers 2–6 months: ∼50-60%.

Techniques: Plasmid Preparation