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. 2019 Oct 10;118(4):967–979. doi: 10.1016/j.bpj.2019.10.006

Figure 7.

Figure 7

Fluorophore-assisted laser inactivation (FALI) of ribbons with a FITC-conjugated RIBEYE-binding peptide indicates that multiquantal release occurs preferentially at ribbon release sites. (A) Confocal z-stack image of a rod after introducing the peptide through a patch pipette is shown. Weak fluorescence from cytoplasmic dye is visible in the outer segment (OS) and inner segment (IS). Two small bright spots in the terminal (arrow) show dye bound to ribbons before laser bleaching. (B) Damaging ribbons by FALI reduced evoked IA(glu) events evoked by a brief depolarizing stimulus (2 ms, −25 mV). The insets show examples of responses evoked in a rod before and after 60-s bleach with 488-nm laser light. (C) FALI with the RIBEYE-binding peptide significantly reduced the frequency of multiquantal events (n = 7 rods; p = 0.0127; two-way ANOVA/Tukey’s multiple comparisons), but FALI with a scrambled control peptide did not (n = 7, p = 0.91; two-way ANOVA/Tukey’s multiple comparisons). (D) FALI did not reduce the frequency of uniquantal events with either the RIBEYE-binding (p = 0.3; 2-way ANOVA/Tukey’s multiple comparisons) or scrambled control peptide (p = 0.84; 2-way ANOVA/Tukey’s multiple comparisons). Data points show mean ± SEM.