Journal: Scientific Reports

Article Title: Cortical diurnal rhythms remain intact with microglial depletion

doi: 10.1038/s41598-021-04079-w

Figure Lengend Snippet: Perineuronal nets increase in a region-dependent manner upon microglial depletion. ( A ) Representative slide-scanned images of WFA (PNN) staining, parvalbumin, and IBA1 immunofluorescence reveal no diurnal changes in PNNs across 24 h and ( B ) a significant increase in PNNs with microglial depletion. ( C ) 10-day PLX5622 (1200 ppm) treatment is sufficient to eliminate >90% of microglia at all timepoints. ( D – G ) Quantification for WFA integrated density for the ( D ) rostral SS Ctx, ( E ) barrel field of the SS Ctx, ( F ) MO Ctx, and ( G ) AUD Ctx show no diurnal PNN differences in control mice and a significant increase in the PLX5622 treated mice. ( H , I ) Quantification of ( H ) WFA+ and ( I ) ACAN+ PNN intensity sum from 20× hippocampal CA1 confocal images show no diurnal differences in PNN intensity nor an increase in PNN intensity with microglial depletion. ( J ) Quantification of ACAN integrated density shows no difference between ZT2 and ZT14, but in the absence of microglia, ACAN integrated density significantly increases. ( K ) DAB staining with ACAN in WT control mice shows no drastic diurnal differences in PFC PNNs, but in ( L ) mice treated with PLX5622, ACAN+ PNN integrated density significantly increases. ( M , N ) Quantification of WFA integrated density in both the ( M ) infralimbic and ( N ) prelimbic area show no diurnal differences in PNNs between ZT6 (mid-light phase) and ZT18 (mid-dark phase). ( O ) Representative slide-scanned images of WFA (PNN) labeling in the PFC. Statistical analysis used a two-tailed unpaired t-test for ILA and PLA WFA Integrated Density quantifications and a two-way ANOVA with Tukey’s multiple comparisons correction the quantifications with PLX5622-treated groups. Significance indicated as *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Primary antibodies used and dilutions are as follows: Iba1 (1:1000, 019–19741, Wako; and 1:500, ab5076, Abcam), Wisteria floribunda agglutinin (WFA) lectin (1:1000, B-1355, Vector Labs), Aggrecan (1:200, AB1031, Millipore), and Parvalbumin (1:500, MAB1572, Millipore).

Techniques: Staining, Immunofluorescence, Labeling, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Cognitively impaired aged Octodon degus recapitulate major neuropathological features of sporadic Alzheimer’s disease

doi: 10.1186/s40478-022-01481-x

Figure Lengend Snippet: Methods use for immunofluorescence marker quantification and analysis

Article Snippet: Perineuronal nets (PNN) were immunostained using biotinylated Wisteria floribunda agglutinin (WFA; Vector Laboratories, cat# B-1355-2).

Techniques: Immunofluorescence, Marker, Activity Assay

Journal: Acta Neuropathologica Communications

Article Title: Cognitively impaired aged Octodon degus recapitulate major neuropathological features of sporadic Alzheimer’s disease

doi: 10.1186/s40478-022-01481-x

Figure Lengend Snippet: Increased perineuronal nets signal coupled with an increase of PV-positive interneurons in the AD-like aged degu brain. Confocal microscopic views of degu coronal brain sections are aligned as indicated and show focused views of perineuronal nets (PNN) (green), parvalbumin (PV) interneurons (red), and neuronal nuclei (NeuN, blue) in entorhinal cortex (EC, a and d ), hippocampal CA2 and CA3 (Hippo, b and e ), and thalamic reticular nucleus (TRN, c and f ). g – l Quantification shows significantly increased counts of PNN(+)-neurons per mm 2 ( g ) and PV(+)-neurons per mm 2 ( h ) in EC ( p = 0.0022 and p 0.0260, respectively) in AD-like degu brains compared to Non-AD controls, along with similar trends for hippocampal CA3/CA2 ( p = 0.0931 for PNN; p = 0.3636 for PV) and TRN ( p = 0.3939 for PNN; p = 0.5887 for PV). Error bars represent SEM; * p < 0.05, ** p < 0.01, n = 6 degus in AD-like and Non-AD groups, n = 3 for male AD-like and female AD-like groups

Article Snippet: Perineuronal nets (PNN) were immunostained using biotinylated Wisteria floribunda agglutinin (WFA; Vector Laboratories, cat# B-1355-2).

Techniques:

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 YA: ∼60-70%.

Techniques: Plasmid Preparation