Figure 6.

Ca²⁺ Responses and Glutamate Inputs in eat-4 Mutants

(A) Averaged glutamate input (blue) and Ca²⁺ response (red) to odor. The shaded region indicates IAA stimulation and the shadows around the solid lines indicate the SEM (N = 12).

(B) Average glutamate intensity before (pre-addition) and after (post-addition) odor stimulation onset (left), and average intensity before (pre-removal) and after (post-removal) odor stimulation removal (right). Error bars indicate the SEM. The statistical metrics are as follows: left, p = 0.41; right, p = 0.055; paired t test (N = 12).

(C) Ca²⁺ spike probability following odor addition. Comparison between N2 animals and glc-3 mutants: p = 0.13; comparison between N2 animals and eat-4 mutants: p = 0.014. Fisher's exact test with Holm correction (N2, N = 15; glc-3, N = 12; eat-4, N = 12); *p < 0.05.

(D) Ca²⁺ spike frequency in N2 (filled), glc-3 (striped), and eat-4 mutants (hollow) with and without odor. Box plots indicate the median (center line), quartiles (boxes), and range (whiskers). The statistical metrics are as follows: N2 data comparison between no odor and later odor, p = 0.0045; glc-3 data comparison between no odor and later odor, p = 0.34; eat-4 data comparison between no odor and later odor, p = 0.34; Wilcoxon signed rank test. No odor data comparison between N2 and glc-3, p = 0.024; no odor data comparison between N2 and eat-4, p = 0.00044; later odor data comparison between N2 and glc-3, p = 0.014; later odor data comparison between N2 and eat-4, p = 0.000015; Wilcoxon rank-sum test. The Holm method was used for the correction (N2, N = 15; glc-3, N = 12; eat-4, N = 12). *p < 0.05, **p < 0.01, ***p < 0.001.

(E) Averaged glutamate responses coincident with Ca²⁺ spikes. Time 0 indicates the onset of the Ca²⁺ spike. All glutamate responses were normalized to zero at time 0. The purple, blue, and green lines indicate the responses under the no odor, early odor, and later odor conditions, respectively. The bars indicate the SEM (N = 12, no odor: n = 9; early odor, n = 3; later odor, n = 9).

(F) Glutamate input before Ca²⁺ spikes and at Ca²⁺ spike onset. “Pre” indicates the average intensity from −3 to −2 s in (E), and “onset” indicates the average intensity from −0.5 to +0.5 s. The statistical metrics are as follows: no odor: p = 0.60; early odor, p = 0.53; later odor, p = 0.50; paired t test (N = 12; no odor, n = 9; early odor, n = 3; later odor, n = 9).

(G) Averaged glutamate decreases (filled boxes) with and without (hollow boxes) Ca²⁺ spikes. Box plots indicate the median (center line), quartiles (boxes), and range (whiskers). The statistical metrics are as follows: no odor, p = 0.88; later odor, p = 0.48; Welch's t test (N = 12, no odor, spike, n = 9; no odor, no spike, n = 108; later odor, spike, n = 9; later odor, no spike, n = 100).