Skip to main content
. 2015 May 20;35(20):8021–8034. doi: 10.1523/JNEUROSCI.0214-15.2015

Figure 8.

Figure 8.

EGFP expression in discrete regions of the CNS of Scn10a-EGFP mice. A, IHC of various brain regions. First row panels represent low-magnification (20×/0.30 NA objective) fluorescence images obtained from the EGFP channel. Scale bar, 50 μm. Remaining images are higher-magnification (63×/1.2 NA objective) fluorescent images from the EGFP channel (second row), red channel (third row), or merged images from both fluorescence channels pseudo-colored green and purple (fourth row). Scale bar, 10 μm for images obtained at higher magnification. B, Conventional RT-PCR analysis of mRNA encoding Scn10a and β-actin in multiple brain regions and peripheral ganglia. Based on the primer sets used, the predicted sizes of the Scn10a and β-actin amplicons are 742 and 500 bp, respectively. The resultant PCR products were separated and visualized on a 1.5% agarose gel. A, Amygdala; B, brainstem; CM, primary motor cortex; CS, somatosensory cortex; G, globus pallidus; Hi, hippocampus; Hy, hypothalamus; O, olfactory tubercle; S, striatum; T, thalamus; D, DRG; SCG, superior cervical ganglia; ND, nodose ganglia; W, water; L, ladder. C, Expression of Scn10a transcript across various brain regions and peripheral ganglia as assessed by qPCR. Relative expression of Scn10a mRNA to DRG tissue was obtained from the formula 2−ΔΔCt (detailed in Materials and Methods). Dotted line indicates mean normalized expression level in DRG neurons (2−ΔΔCt = 1). Each point indicates a biological replicate. Means are represented as bars. Error bars indicate SEM. The differences between mean 2−ΔΔCt values for all regions with DRGs were statistically significant, except for nodose (p < 0.01, one-way ANOVA with Holm-Sidak's multiple-comparisons test). D, Acutely prepared coronal brain slice under fluorescence optics. Hypothalamic region magnified to the right. Ei, Acutely dissociated hypothalamic neurons under phase-contrast (above) or EGFP fluorescence (below). Eii, Sample electrophysiological recording from an EGFP-positive dissociated hypothalamic neuron. Black trace represents INa recorded in the presence of 3 μm TTX. Red trace represents INa recorded in the presence of 3 μm TTX and 3 μm A-803467 drug. Four consecutive current traces for each condition were averaged and represented.