Journal: Frontiers in Molecular Neuroscience

Article Title: A localized tracing technique to explore intra-amygdala functional and structural correlates of individual variability in behavioral response

doi: 10.3389/fnmol.2025.1347539

Figure Lengend Snippet: Structural and functional connectivity patterns within the amygdala complex do not mediate individual variability in stress response. (A) Viral tracing and fluorescent labeling scheme show colors associated with each additional fluorescent marker (red = TVA-mCherry-G, green = oG-rabies-GFP, magenta = cFos). (B) Example triple-channel image of the amygdala region (red = mCherry, green = GFP, magenta = cFos). Top inset (pink box and outline) shows examples of a structural projection neuron (open pink arrow, GFP + cFos − ) and a functional projection neuron (closed pink arrow, GFP + cFos + ). Bottom inset (yellow box and outline) shows examples of a starter neuron (closed yellow arrow, GFP + mCherry + ) and a projection neuron (open yellow arrow GFP + mCherry − ). (C) Map of all identified starter cells (GFP + mCherry + ). (D) Correlations between starter cell count in each region and SI ratio with FDR-corrected and non-FDR-corrected significance threshold indicated (solid horizontal line, Pearson R = 0.835, p = 0.00263; dashed horizontal line, Pearson R = 0.632, p = 0.05; respectively). Starter cell counts did not predict behavioral phenotype in any identified brain region. (E) Map of all identified structural (GFP + cFos − ) and functional (stress-activated) projection (GFP + cFos + ) neurons. (F) Correlations between structural projection neuron (GFP + cFos − ) count (striped) and functional projection neuron (GFP + cFos + ) count (solid) in each region and SI ratio with FDR-corrected and non-FDR-corrected significance threshold indicated (solid horizontal line, Pearson R = 0.749, p = 0.0125; dashed horizontal line, Pearson R = 0.632, p = 0.05; respectively). No significant correlations identified. (G) Correlations between overall number of activated cells (cFos + ) per brain region and SI ratios.

Article Snippet: Sections were serially stained with rabbit anti-cFos (9F6) primary monoclonal antibody (Cell Signaling, catalog number: 2250S, lot number: 12, 1:2,000 in 0.3% PBST + 5% Normal Goat Serum + 1% BSA + 0.2% Cold Water Fish Skin Gelatin block solution) and Alexa Fluor-conjugated AffiniPure goat anti-rabbit secondary antibody (Alexa Fluor-647, Sigma, code number: 111-605-033, lot number: 157045, 1:500 in 0.3% PBS-T).

Techniques: Functional Assay, Labeling, Marker, Cell Counting