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Figure 11
Shh regulates cerebellar proliferation and differentiation following injury. (A) Expression of Shh pathway genes by qRT-PCR reveals that the shha and shhb mRNAs are upregulated by 4 hpi in the top 1/3rd of the cerebellum, while gli and smo expression increased by 12 hpi (5 pooled cerebellums/group, n = 3 groups). (B?G) Coronal cerebellar sections of the CC of undamaged (B?D) and sTBI fish (E?G) that were IP-injected with EdU and either vehicle (C,F), the Smo agonist purmorphamine (D), or the Smo antagonist cyclopamine (G). (H,H??I,I?) Coronal cerebellar sections of undamaged fish that were either untreated (H?H??) or purmorphamine-treated (I?I??) that were IP-injected with EdU and collected at 7 dpi. Coronal cerebellar sections of sTBI fish that were either untreated (J?J??) or cyclopamine-treated (K?K??) that were IP-injected with EdU and collected at 7 dpi. (L) Quantification of the number of EdU-positive cells at the CC in undamaged and sTBI fish with Shh modulation (n = 9). (M,N) Quantification of the number of EdU/HuCD double-positive cells in the granule cell layer of the cerebellum in undamaged and sTBI fish with Shh modulation (n = 10). Solid lines in (B?K?) denote tissue boundary, while dotted lines denote internal anatomical boundaries. Cerebellar crest, CC, granule cell layer, GL, molecular layer, ML. All scale bars = 100 µm. Mean ± SEM is depicted in (L?N). Statistical analyses were performed with either a One-way ANOVA or Two-way ANOVA followed by a Tukey?s post-hoc test. ## p < 0.01.