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Fig. 3.
Localization of TRPN1 in Xenopus vestibular hair cells. (A) Saccular hair cells in profile. Triple staining for TRPN1 (green), actin (red), and tubulin (blue); pseudocolor code also applies to B, E, F, K, and O. Arrows indicate TRPN1 at the kinocilial bulb. (B) Preincubation of the TRPN1 antibody with the epitope peptide prevents TRPN1 staining at the kinocilia tip. (C and D) Cdh23 (green), actin (red), and tubulin (blue) in hair cell under control conditions. Cdh23 is present in the two to six tallest stereocilia in a stripe adjacent to the kinocilial bulb. (E) TRPN1 and actin in hair cell under control conditions. Note TRPN1 in kinocilia bulb (arrow). (F) After EGTA treatment (5 mM, 30 min), the TRPN1 signal appears at the base of the kinocilium (arrow). (G) Graphic illustration of the optical section in the confocal sections of H-O. (H-O) Whole-mount view of hair bundles, in a slightly angular optical section as illustrated in G. (H-K) Control conditions. (H) Actin. (I) TRPN1. (J) Tubulin. (K) Merged signals. (L-O) After EGTA treatment. (L) Actin. (M) TRPN1. (N) Tubulin. (O) Merged signals. Full panel widths: 24 μm, A and B; 8 μm, C; 25 μm, D; 13 μm, E and F; 45 μm, H-O. |
