Fig. 4: Glutamate release from ASER is reduced in pkc-1(nj3) and unc-64(S65A).

a A schematic illustration of the fluorescent reporter of glutamate release, eat-4::pHluorin, used in B and C²². Release of glutamatergic synaptic vesicles causes increased fluorescence. b Glutamate release is reduced in pkc-1(nj3) mutants. n = 20 independent animals. c Glutamate release is reduced in unc-64(S65A) mutants. n = 20 independent animals. p values were calculated by the two sided Welch’s test (single comparison) or Tukey’s test (multiple comparisons). The gray vertical shade in b, c shows the decrease in NaCl concentration applied to the worm nose (50–25 mM). In b, c, left, the lines with the shade show the average and SEM, respectively, of changes in pHluorin fluorescence, and in b, c, right, the boxes extends from 25th to 75th percentile, median is marked by the line, and whiskers indicate the minimum and the maximum values in 1.5× the inter-quartile range. d A schematic illustration of the FRAP of ASERp::eat-4::pHluorin. EAT-4::pHluorin was photobleached using high-intensity 488 nm Argon laser until the fluorescence decreased to ~50% of the original level. Once fluorescent EAT-4::pHluorin at the surface membrane is photobleached, the fluorescence is recovered according to the release of synaptic vesicle containing EAT-4::pHluorin. Since worms do not experience any change in NaCl concentration during image acquisition, this recovery in fluorescence reflects the baseline release of glutamate. e The result of FRAP in wild-type animals, pkc-1(nj3) and unc-64(S65A) mutants (n = 8 independent animals). Error bars indicate SEM. The time course of fractional recovery was linearly fitted. p value were calculated by t values of interaction between time course and strains in generalized linear model with Bonferroni’s correction. Source data are provided as a Source Data file.