Figure 4.

Figure 4.

Casper zebrafish larvae were first pretreated at 60 hr post-fertilization (hpf) with either vehicle control (A and B), 0.03 mM dopamine (C), or 0.03 mM L-mimosine (D). Larvae were then rinsed at 72 hpf and treated with either vehicle control (A) or 0.01 mM cisplatin B-D). Two days later (48 hr cisplatin treatment), larvae were fixed, permeabilized, then stained with AlexaFluor488-Phalloidin, then imaged with a Zeiss LSM510 confocal microscope. Larvae mounted laterally with ventral side at the top. Scale bar = 20 µM. N=6/treatment. White line dashed represents the average posterior macula placode of three control larvae, red dashed line represents this average area superimposed onto other treatments. Note that these images were taken with the exact same settings to ensure accurate representation. (E) To perform topological analysis of the hair cells, the original Zeiss LSM files of 2D images ranging from 27 to 42 μm in depth were imported to Imaris v.X64 9.1.2 software for surface reconstruction. Ellipticity (prolate, or elongation around the long axis) was used as a measure of the status of the hair cells with healthier hair cells exhibiting a higher prolate ellipticity. **=p<0.01, ****=p<0.001, as per Kruskal-Wallis testing with a Dunn’s multiple comparison test. (F) Hair cell numbers were counted using the Cell Counter plugin in ImageJ using the same maximum projection images used in E). Individual points represent the number of hair cells/larvae. *=p<0.05, **=p<0.01, and ****=p<0.001, as per one-way ANOVA with a Tukey post-test. N = 6 larvae/treatment with 16 hair cells measured/larvae.