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. 2023 Jul 14;14:4218. doi: 10.1038/s41467-023-39955-8

Fig. 3. A PKA signaling cascade functions in hmc to facilitate hmc activation.

Fig. 3

a Quantification of the number of aBocs per cycle in adult animals of the indicated genotypes. “hmc PKA[DN]” denotes a transgene in which a PKA dominant negative variant is expressed in hmc using the nmur-3(Δ) promoter. RNAi denotes RNA interference-mediated knockdown of the indicated gene by feeding. Data are presented as mean values ± SEM. n = 6, 3, 9, 3, 4, 2 independent animals. ***P < 0.001 in one-way ANOVA with Dunnett’s correction for multiple comparisons; n.s. not significant. b Quantification of the number of hmc calcium spikes observed per cycle in adult animals of the indicated genotype. “hmc PKA[DN]” and “hmc PKA[CA]” denote transgenes expressing PKA dominant negative or PKA constitutively active variants in hmc under the control of the nmur-3(Δ) promoter. For (b) and (e), wild-type: 33 cycles in 8 animals, hmc PKA[DN]: 63 cycles in 21 animals, frpr-17: 56 cycles in 13 animals, frpr-17; hmc PKA[CA]: 27 cycles in 6 animals. ***P < 0.001 in two-sided chi-square test with Bonferroni’s correction for multiple comparisons; n.s. not significant. c Violin plots of calcium spike initiation time in hmc after the end of the intestinal calcium oscillation. Dashed lines refer to median and dotted lines refer to quartiles. ***P < 0.001 and *P < 0.05 in Kruskal–Wallis test with Dunn’s correction for multiple comparisons. d Left: average traces of calcium dynamics in hmc aligned to the calcium spike initiation time in the indicated mutants. “hmc PKA[DN]” and “hmc PKA[CA]” denote transgenes expressing PKA dominant negative or PKA constitutively active variants in hmc under the control of the nmur-3(Δ) promoter. The solid lines indicate average fold change in GCaMP intensity and the shades indicate SEM. Dotted line represents a possible critical threshold for aBoc. Right: quantification of the average peak amplitude and half-decay time. Data are presented as mean values ± SEM n = 10, 7, 5 cycles from different animals. ***P < 0.001 and **P < 0.01 in one-way ANOVA with Bonferroni’s correction for multiple comparisons and in two-tailed Student’s t-test. e Quantification of the number aBocs observed per cycle in the indicated mutants in the time lapse calcium images. “hmc PKA[DN]” and “hmc PKA[CA]” denote transgenes expressing PKA dominant negative or PKA constitutively active variants in hmc under the control of the nmur-3(Δ) promoter. ***P < 0.001 in two-sided chi-square test with Bonferroni’s correction for multiple comparisons; n.s. not significant.