DEV012237 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. eya4 expression during otic placode and otic vesicle development. (A-C) At 14, 16 and 18 hpf, wild-type embryos were hybridized to DIG-labeled eya4 antisense RNA probes. Lateral views of the otic placode region are shown. (D,E) Sections from wild-type embryos were hybridized with a DIG-labeled eya4 antisense RNA probe. eya4 signal (arrows) at 24 hpf (D) demarcated the developing sensory epithelia (SE) and neuroblast (NB). A section from 72 hpf (E) demonstrated eya4 expression in the anterior maculae (AM), anterior cristae (AC) and posterior cristae (PC) of the sensory epithelia. D, dorsal; V, ventral; A, anterior; P, posterior.

• Supplemental Figure S2 -

  Fig. S2. Acridine Orange staining of otic vesicles from 24 and 48 hpf wild-type, eya4 and atp1b2b morphant fish. Positive signals (indicated by white arrows) were observed in a small number of embryos. Those embryos with maximal staining from each group at both time points are presented. In general, only background levels of Acridine Orange staining were observed in wild type, and eya4 and atp1b2b morphants.

• Movie 1 -

  Movie 1. Three-dimensional reconstruction of the otic vesicle surface in wild-type zebrafish. This and other movies are also available at http://genepath.med.harvard.edu/~seidman/movies/20070713-atp1b2b/index.html

• Movie 2 -

  Movie 2. Three-dimensional reconstruction of the otic vesicle surface in eya4 morphant fish.

• Movie 3 -

  Movie 3. Representative startle responses in wild-type fish (72 hpf). Startle responses were elicited by tapping the 100-mm petri dish. Responses of individual zebrafish (one embryos per dish) were assessed.

• Movie 4 -

  Movie 4. Representative startle responses in eya4 morphant fish (72 hpf). Note circular and uncoordinated swimming patterns.

• Movie 5 -

  Movie 5. Representative startle responses in atp1b2b morphant fish (72 hpf). Note that most fish remain stationary, although some had circular and uncoordinated swimming patterns.

• Movie 6 -

  Movie 6. Representative startle responses in atp1b2b mRNA-treated eya4 morphant fish (72 hpf). Coordinate swimming in response to stimuli is evident.