Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: In vivo quantitative proteomics of somatosensory cortical synapses shows which protein levels are modulated by sensory deprivation

doi: 10.1073/pnas.1300424110

Figure Lengend Snippet: SynGAP1 protein distribution differs in coronal sections from sensory-deprived (trimmed) and sensory-normal (brushed) barrel cortex. (A and B) Immunohistochemistry of whole-hemisphere brain sections through cortex from a representative trimmed (A) and brushed (B) barrel field stained for SynGAP1 alone (Left) and overlaid with VGlut2 staining to mark layer IV (red fluorescence) (Right). (C and D) SynGAP1 immunoreactivity in sections from A (trimmed, red) and B (brushed, blue) was graphed as a function of cortical depth through layer I (C) and layer IV (D). (E) The ratio of total layer I to layer IV SynGAP1 staining was reduced significantly in sensory-deprived (trimmed) vs. sensory-normal (brushed) barrel cortex (P = 0.012, two-tailed paired t test, n = 4 animals per condition). Total layer I to layer IV SynGAP1 immunoreactivity ratios were calculated by dividing the average immunoreactivity across layer I by the average immunoreactivity across layer IV for a given tissue section, and these values were averaged across four animals per condition. This representation of the staining ratio provides an internal normalization so that staining in different sections can be compared. SynGAP1 immunoreactivity across all cortical layers for all animals represented here is graphed in Fig. S1.

Article Snippet: Sections were imaged on a fluorescence scanner (NanoZoomer; Hamamatsu) with transmitted light and Texas Red filter sets to image 3,3'-Diaminobenzidine and Alexa Fluor 594, respectively.

Techniques: Immunohistochemistry, Staining, Fluorescence, Two Tailed Test