Figure 5.

Vital imaging can define areas of neuronal OE regeneration after lesioning

(A) Whole mount staining of the nasal cavity from a methimazole/dichlobenil lesioned mouse using PGP 9.5/TUBB4 identifies areas on both the lateral wall and septum where respiratory metaplasia (arrowheads) in previous neuronal OE. D = dorsal, V = ventral, A = anterior, p = posterior, OB = olfactory bulb. Scale bars = 2 mm.

(B) A coronal section of the nasal cavity from a methimazole/dichlobenil lesioned mouse demonstrates respiratory metaplasia in the dorsal area of OE (arrows). The arrowhead indicates intracavitary excrescences from the injured OE. Scale bar = 1 mm.

(C) Regenerated ventral OE from the boxed area labeled “C” in Figure 5B contains a thick layer of TUJ1 labeled OSNs without TUBB4+ cells. Scale bar = 25 μm.

(D) A dorsal region of OE from the boxed area labeled “D” in Figure 5B is without neurons and is labeled for respiratory cells with TUBB4 antibodies. Scale bar = 25 μm.

(E) A cartoon illustrates wholemount staining in a methimazole/dichlobenil-injected mouse with respiratory metaplasia in the dorsal recess of the nasal cavity.

(F) Fluorescent imaging of whole mounts incubated with 10 mM coumarin captured before and 30 min after probe application reveals signal in the ventral area of OE but not in dorsal areas of what was formerly OE (arrowheads).

(G) Relative fluorescence intensity of dorsal vs. ventral mucosa measured every 5 min after application of coumarin. The data are shown as the mean ± SEM (n = 5).

(H) Fluorescent imaging of whole mounts incubated with 10 μM Glu-HMRG captured before and 30 min after probe application again reveals signal in the ventral area of OE but not the dorsal area (arrowheads).

(I) Relative fluorescence intensity of dorsal vs. ventral mucosa measured every 5 min after application of gGlu-HMRG. The data are shown as the mean ± SEM (n = 5). D = dorsal, V = ventral, A = anterior, p = posterior, OB = olfactory bulb, scale bars = 2 mm.