Fig. 4.

Endogenously produced betaine is required to promote global search and food leaving behaviors. Behavioral responses of N2 (wild-type), alh-11(lj118), alh-11(lj118); alh-11p::alh-11, and alh-11(lj118); gcy-13p(RIM)::alh-11 worms. (A) Schematic representation of the experimental design and (B) Tukey’s boxplot of dispersal assay, in which the zone density ratio of the central zone vs. the whole plate is calculated and plotted. N = 3–29 plates per genotype. (C) Representative images of threshold-filtered plates used to calculated dispersal index. (D) Schematic representation of the experimental design and (E) Tukey’s box plot of food leaving probability 6 h after being placed on the food patch. N = 16–21 plates per genotype. (F) Schematic representation of the experiment (G) Chemotaxis index toward 0.5% diacetyl. N = 3–5 plates per genotype presented in a box and whisker plot. (H) Schematic representation of the experimental design (I) Amount of reorientation events off food at three different time points, as measured per worm per minute. N = 80–157 per genotype. Bars show median value with 95% CI and significance test using the Kruskal–Wallis test *P < 0.05, **P < 0.005, ***P < 0.0005. (J) Overlaid trajectories of individual alh-11 mutant animals at 0–5 min and 40–45 min after removal from a food patch. 0–5 min: N = 69, 40–45 min: N = 109 individual worms. (K) Schematic outlining workflow for transporter assay in Xenopus oocytes. (L) ³H-betaine uptake during 10 min evaluated in the C. elegans genes unc-17, cat-1, T28F3.4, ZK54.1a, and ZK682.2 as well as the human betaine/GABA transporter BGT1 (SLC6A12). Error bars represent SEM of 6–10 samples per genotype, **P < 0.01, ****P < 0.0001, calculated by a one-way ANOVA using Sidak’s test for multiple comparisons. (B, C, E, and G) Tukey’s boxplots showing the median value and a one-way ANOVA with Bonferroni (B and G) or Tukey’s (C and E) correction for multiple comparisons, **P < 0.005, ***P < 0.001, ****P < 0.0001.