FM 1-43 photo-oxidation suggests that synaptic vesicles recycle at the base of IHCs.


<p>(<b>A</b>)–(<b>D</b>) The electron micrographs show organelles containing the photo-oxidation product (DAB precipitate) at rest (<b>A</b>), during stimulation (<b>B</b>), or after a 5-minute (<b>C</b>) or 30-minute (<b>D</b>) recovery period. Some typical organelles are indicated by red arrowheads. Large tubular organelles are indicated by the dashed red traces. Mitochondria are indicated by white asterisks. Note that the basal region contains few labeled organelles at rest; their number increases after stimulation. Note also that the tubular organelles from the top and nuclear regions, found both at rest and during stimulation, disappear during the recovery periods. Scale bars, 200 nm. (<b>E</b>–<b>H</b>) Analysis of organelle labeling under the different experimental conditions. The percentage of the cell volume occupied by labeled organelles was measured in four different cellular regions: cuticular plate (<b>E</b>), top (<b>F</b>), nuclear (<b>G</b>) and basal (<b>H</b>). The graphs show averages performed for electron micrographs from 14 (resting), 12 (stimulation), 4 (5 minute recovery) and 4 (30 minute recovery) independent experiments. We used one-way ANOVA tests to check whether stimulation produced any changes in the different cellular regions. No significant differences were found for the cuticular plate region (<b>E</b>), for the top region (<b>F</b>), and for the nuclear region (<b>G</b>); all <i>P</i> values were higher than 0.05. In contrast, the ANOVA test revealed significant differences for the basal region (<b>H</b>); <i>P</i><0.05. A <i>post hoc</i> Bonferroni test indicated that stimulation increases the amount of label significantly in this region, compared to the resting control (<i>P</i><0.001).</p

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