Fig. 6: Itsn1 is required for glutamate release and synaptic facilitation.
From: Intersectin and endophilin condensates prime synaptic vesicles for release site replenishment
a, Representative images of Itsn1 WT (top) or Itsn1 KO (bottom) neurons expressing iGluSnFR3 v857. Left images show neuronal processes at baseline. Middle images show neurons directly after a 1-ms e-pulse was applied to cells. Right shows the same regions transformed to show peak dF/F values. Synaptic regions of interest (ROIs) are shown as a dotted circle. Scale bar, 10 µm. b, Peak iGluSnFR dF/F values resulting from glutamate release from a single stimulation in either Itsn1 WT or Itsn1 KO neurons. Bars are the mean; error bars are s.e.m. Two-sided Mann–Whitney test. ****P < 0.0001. c, Peak iGluSnFR dF/F values resulting from glutamate release from the second stimulation in a paired-pulse experiment. Paired-pulse intervals range from 50 ms to 1,000 ms (1 s). Bars are the mean; error bars are s.e.m. Kruskal–Wallis test, with Dunn’s multiple comparisons test. ***P = 0.0003 and ****P < 0.0001. Comparisons were made between Itsn1 WT and KO cells at 50 ms, 200 ms and 1,000 ms, respectively. d, PPR plotted for paired-pulse experiments conducted at varying paired-pulse intervals, ranging from 50 ms to 1,000 ms (1 s). PPRs are quantified as the peak dF/F value from stimulation 2 (P2) divided by the peak dF/F value from stimulation 1 (P1). Dots are the mean; error bars are s.e.m. Kruskal–Wallis test, with Dunn’s multiple comparisons test. ****P < 0.0001. Inset, representative dF/F traces from either Itsn1 WT or Itsn1 KO cells taken from a paired-pulse experiment with a 50 ms time interval. Lines are an average of all release site responses; error bars are s.e.m. See Supplementary Table 1 for additional information.
