daf-18/PTEN mutants are hypersensitive to cholinergic drugs.
A. Schematic representation of paralysis induced by aldicarb and levamisole. Aldicarb acts by inhibiting acetylcholinesterase, leading to an accumulation of acetylcholine at the neuromuscular junction, resulting in continuous stimulation of muscles and worm paralysis. Levamisole functions as an agonist at nicotinic acetylcholine receptors, causing prolonged depolarization and, also, muscle paralysis. B-E. Quantification of paralysis induced by aldicarb (Top) and levamisole (Bottom). The assays were performed in NGM plates containing 2 mM aldicarb or 0.5 mM levamisole. Strains tested: N2 (wild-type) (B-E), CB1375 daf-18(e1375) (B), OAR144 daf-18(ok480) (B-C), GR1310 akt-1(mg144) (D), TJ1052 age-1(hx546) (D), VC204 akt-2(ok393) (D), VC222 raga-1(ok386) (E) and KQ1366 rict-1(ft7) (E). All of these strains carry loss-of-function mutations. Furthermore, the strains denoted as "pdk-1 (lf)" and "(gf)" correspond to JT9609 pdk-1(sa680), which possesses a loss-of-function mutation, and GR1318 pdk-1(mg142), which harbors a gain-of-function mutation in the pdk-1 gene, respectively. The strain CB156 unc-25(e156) was included as a strong GABA-deficient control (B-D). At least four independent trials for each condition were performed (n= 25-30 animals per trial). One-way ANOVA was used to test statistical differences in the area under the curve (AUC) among different strains. Post-hoc analysis after One-Way ANOVA was performed using Tukey’s multiple comparisons test (B and C) and Dunnet’s to compare against the wild-type strain (D and E) (ns p > 0.05; * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001).