Journal: Frontiers in Cell and Developmental Biology
Article Title: Pharmacologic Inhibition of ADAM10 Attenuates Brain Tissue Loss, Axonal Injury and Pro-inflammatory Gene Expression Following Traumatic Brain Injury in Mice
doi: 10.3389/fcell.2021.661462
Figure Lengend Snippet: GI254023X reduces axonal injury and alters the gene expression of glutamate receptor subunits after TBI. (A) Representative western blot showing TBI-induced generation of spectrin breakdown products (SBDPs, 145–150 kDa) at 7 dpi in ipsilesional brain tissue samples. (B) Semi-quantitative densitometry of protein bands revealed reduced levels of SBDPs in GI254023X-treated animals compared to vehicle treatment ( n = 11 each). (C) Immunofluorescence images of cryosections for non-phosphorylated Neurofilament-H (SMI-32) at 7 dpi at perilesional sites indicate increased axonal injury at 7 dpi in CCI animals compared to sham. (D) Quantitative assessment of SMI-32 immunoreactive (IR) particles at 7 dpi showing decreased numbers of SMI-32-IR particles in GI254023X- compared to vehicle-treated animals ( n = 12 each). (E–G) Histograms showing relative mRNA expression of AMPAR1 ( Gria1 ), NMDAR2B ( Grin2b) , and CamK2A ( Camk2a ) at 7 dpi in ipsilesional brain tissue samples. Expression values were normalized to the reference Ppia (cyclophilin A) and data from CCI animals ( n = 12 each group) are shown relative to sham (red line = sham). Two outliers were removed after identification by ROUT’s test in (B) . Values from individual animals and mean ± SEM are shown. Differences between GI254023X- and vehicle-treated animals were calculated by Student’s t -test (* p < 0.05, ** p < 0.01, *** p < 0.001).
Article Snippet: Nitrocellulose membranes were washed in TBS containing 0.1% Tween, blocked with 2.5% non-fat milk and incubated with mouse anti-α-fodrin/αII-spectrin (Enzo Life Sciences, 1:750, RRID:AB_10554860 ), mouse anti-GAPDH (Acris Antibodies GmbH, 1:1,000, RRID:AB_1616730 ), and secondary infrared (IR) dye-conjugated antibodies (LI-COR; goat anti-mouse IgG IRDye 800, RRID:AB_10793856 ).
Techniques: Expressing, Western Blot, Immunofluorescence
