Fig. 3.

Intracellular localization of AhR in mouse LC-NA neurons at P5, P7, and P14. a Immunostained LC-NA neurons double-positive for TH and AhR. The cellular boundary of TH-stained cells was clearly observed, which enabled the measurement of the nuclear and soma areas of individual LC-NA neurons. Scale bar = 10 μm. b–f Quantitative analyses of AhR-expressing LC-NA neurons. The nuclear area was significantly larger at P5 than at P7 and P14 (b), whereas the soma area was significantly larger at P14 compared to P5 and P7 (c), indicating that marked changes in cellular morphology occur between P5 and P14. The nuclear area percentage that represents the nuclear area normalized to the soma area in each neuron was significantly decreased during the period from P5 to P14 (d). The AhR immunostaining intensity per nucleus (AhR^Nuc intensity) was normalized to that of the soma in each neuron (AhR^Nuc intensity percentage). The AhR^Nuc intensity percentage at P5 was significantly higher than that at P7 and P14 (e). To normalize the developmental-stage-related changes in cellular morphology, the ratio calculated by dividing AhR^Nuc intensity percentage by nuclear area percentage (ratio^LC−NA) served as an index to evaluate the nuclear AhR. No significant difference in ratio^LC−NA was found across developmental periods (f). Values are shown as the mean ± SD. Circles represent individual cell data (215, 228, and 200 cells from 3 mice each at P5, P7, and P14, respectively). Asterisks (** and ***) denote statistical significance at p < 0.01 and 0.001, respectively, by one-way ANOVA with the Tukey–Kramer post hoc test