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Figure 8. Kv2.2 protein was detected in the developing otic vesicle and the hindbrain. Transverse cryostat (18 μm) sections were incubated with the Kv2.2 antibody or preblocked Kv2.2 antibody (inset). Sections were counterstained with Hoechst (blue). A: At St 41, Kv2.2 immunoreactivity was present in the ventral lateral hindbrain and the medial epithelium of the otic vesicle (asterisk), where the sacculus forms. B: A magnified image of the boxed area in A illustrates the localization of Kv2.2 protein to the medial side of the otic epithelium. Kv2.2 protein was also detected in the innervating nerve (arrowhead) and the spiral ganglion cells (sg). C: At St 44, the Kv2.2 immunolabeling in the hindbrain had expanded both dorsally and medially. Kv2.2 protein was again detected in the saccular portion of the otic epithelium. D: A magnified view of the boxed area in C shows Kv2.2 protein again at the medial side of the otic epithelium. Also, Kv2.2 immunolabeling persisted in the innervating nerve and the spiral ganglion cells. Inset: Preincubation of the Kv2.2 antibody with the Kv2.2 peptide block effectively eliminated Kv2.2 immunolabeling. E: At St 50, Kv2.2 protein expression had continued both dorsal and medial expansion in the hindbrain. The Kv2.2 immunolabeling in the otic epithelium was again present. F: A magnified view of the boxed area in D illustrates a strong Kv2.2 presence in the otic epithelium as well as the spiral ganglion cells and innervating nerve. In the inset, Kv2.2 immunolabeling is detected around the membrane of the developing hair cell, with large cytosolic portions of the protein evident toward the apical portion of the soma. nc, Notochord; ov, otic vesicle; rp, roof plate; sg, spiral ganglion. Scale bars = 200 μm in A,C,E; 20 μm in B; 20 μm in F (applies to D,F), 5 μm for inset. |
