Figure 6. Intact and zip-code-less TH mRNAs are locally translated, enhance axonal levels of TH proteins and facilitate presynaptic NE synthesis. A, Schematic representation of the Campenot culture chambers, the sites of mRNA transfection are indicated with arrows. B, Intra-axonal and soma TH and NE levels were measured 1 d after transfection, using immunocytochemistry in soma and axons, in which axons were lipofected with intact or zip-code-less TH mRNA or sham transfected. Increased levels of TH and NE are detected in axons locally overexpressing TH, whereas the soma levels of TH, and NE remained at control levels. C, D, Fluorescence intensity as a measure of TH (C) and NE (D) was quantified in the axons and soma of SCG neurons using ImageJ. Fluorescence levels are provided as relative fluorescence intensity. Data are mean ± SEM from the measurement of 16–22 axons and 16–18 SCG ganglia from three independent experiments. One-way ANOVA, ***p ≤ 0.0001. E, Schematic representation of the Campenot culture chambers, the site of mRNA transfection in the center compartment is indicated with an arrow. F, Intra-axonal and soma TH and NE levels were measured 1 d after transfection, using immunocytochemistry in axons and cognate soma, in which soma were lipofected with intact or zip-code-less TH mRNA. Sham-transfected neurons served as negative controls. Increased levels of TH and NE are detected in soma and proximal axons locally overexpressing TH, whereas the distal axon levels of TH, and NE remained at control levels. G, H, Fluorescence intensity as a measure of TH (G) and NE (H) levels was quantified in the axons and soma of SCG neurons using ImageJ, and fluorescence levels are provided as relative fluorescence intensity. Data are the mean ± SEM from the measurement of 16–22 axons and 16–18 SCG ganglia from three independent experiments. One-way ANOVA, ***p ≤ 0.0001.