Fig. 3. Ciliogenesis in POMC neurons during the early postnatal period is critical for adulthood energy balance.
a Cilia (AC3) staining in the developing hypothalamus of C57 mice (n = 4 for P0 and P28, n = 5 for E12.5, E15.5, P14, and P60, n = 6 for E18.5 and P7). The graphs depict the average length of about 100 ARH cilia and the ciliated cell percentage (the cilia numbers divided by the DAPI numbers) in each mouse. Scale bars: 20 μm. 3V third cerebroventricle. b IFT88 immunoblotting in the developing hypothalamus of C57 mice (n = 4). c AC3 and β-endorphin double staining in the ARH of C57 mice showing POMC neuron ciliogenesis during development (n = 4 for E18.5, P7, and P60, n = 5 for P1 and P14). Scale bars: 20 μm. d Experimental scheme in mice with POMC-specific IFT88 depletion that was induced by tamoxifen injections during the postnatal period (P1–P14). e Representative cilia images showing ciliary dysgenesis in POMC neurons (n = 4). Arrowheads indicate cilia of POMC neurons. Scale bars: 20 μm. f Body weights in mice with postnatal ciliary dysgenesis in POMC neurons and their IFT88f/f litters on a chow diet (n = 5 for males, n = 6 for females). g Lean mass and fat mass measured at 15 weeks (n = 5 for males, n = 6 for females). h Average values of weekly food intake during 10–15 weeks (n = 5 for males, n = 6 for females). i Energy expenditure measured at 12 weeks (n = 4). Data are presented as the mean ± SEM values. Statistics were performed using one-sided one-way ANOVA (a–c), one-sided two-way ANOVA (f) followed by post hoc LSD test and two-sided Student’s t test (e, g–i). *p < 0.05, **p < 0.01, ***p < 0.001 vs. IFT88f/f controls or between the indicated groups.