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. 2022 Feb 16;11(4):684. doi: 10.3390/cells11040684

Figure 5.

Figure 5

L-T4 treatment modulates cell proliferation and neurogenesis after UVN. (a) Study design used to assess cell proliferation and survival of the newly generated cells after unilateral vestibular neurectomy (UVN) and L-thyroxine (L-T4) treatment. UVN-T4 group received L-T4 injections (10 µg/kg, i.p.) and the UVN-NaCl group received 0.9% NaCl (vehicle, in equivalent volume, i.p.) at the end of the UVN and one injection/day during the first 3 days post-lesion. At day 3, animals received a BrdU injection (200 mg/kg, i.p.) and were perfused either 3 h later to evaluate cell proliferation, or on day 30, to investigate survival and differentiation of the newly generated cells. (b) Confocal immunostaining images of BrdU+ cells in the deafferented medial vestibular nucleus (MVN) before the lesion (pre-op), three days (D3) and thirty days (D30) after UVN for UVN-NaCl and UVN-T4 groups. Scale bar = 25 µm. (c) Quantitative assessment of the effect of UVN and L-T4 treatment on the number of BrdU+ cells in the deafferented MVN of UVN-NaCl (red, n = 4/delay) and UVN-T4 (blue, n = 4/delay) groups. (*** p < 0.001; one-way ANOVA and Tuckey’s post hoc tests). (d) Maximum intensity projection of Z-stack confocal images of cell differentiation evaluated in the deafferented MVN 30 days after UVN. The BrdU nuclei are in red, and the other markers of differentiation are: GFAP (astrocyte), IBA1 (microglia), Olig2 (oligodendrocytes), and NeuN (neuron) are in green. Scale bar = 10 µm. (e,f) Proportions of new neurons (green), astrocytes (blue), microglia (orange), and oligodendrocytes (purple) generated in the deafferented MVN of the UVN-NaCl (e) and UVN-T4 (f) groups 30 days after the lesion.