Fig. 1: LGC-46 is a key component of a postsynaptic acetylcholine (ACh) receptor in D-MNs.

a Diagram depicting major components of the forward and backward circuits, and their synaptic connections. A- and B-type cholinergic motor neurons are labeled A and B, respectively, whereas GABAergic motor neurons are labeled D. AVA and AVB are premotor command interneurons for backward and forward movements, respectively. b Spontaneous postsynaptic currents (sPSCs) in VD5 (held at −60 mV) depended on LGC-46 and ACh release. unc-17 and lgc-46 were knocked down in cholinergic neurons using Punc-17 and GABAergic neurons using Punc-47, respectively. c Whole-cell current caused by exogenous ACh (1 mM) in VD5 (held at −60 mV) depended on LGC-46. Compared with wild type (wt), p = 0.000 lgc-46 RNAi, 0.000 lgc-46(ok2949), 1.000 ok2949 rescue, 0.000 lgc-46(ok2900), 0.951 ok2949 rescue. In b and c, GABAergic neuron (D-MN)-targeted rescue was achieved by using Punc-47. d Optogenetically evoked PSCs in VD5 depended on LGC-46. The experiments were performed with strains expressing channelrhodopsin-2 in cholinergic neurons (using Punc-17) either in the presence or absence of all-trans retinal. Compared with wt, the p values for evoked PSC amplitude and evoked PSC charge transfer are 0.000 for both RNAi and wt retinal (−). e Localization of LGC-46::GFP expressed in D-MNs and TagRFP::ELKS-1 (a presynaptic marker) expressed in cholinergic MNs in the dorsal nerve cord. The displayed images represent >20 transgenic worms. Arrows indicate co-localization of the two fusion proteins. Scale bar = 10 µm. f Comparison of locomotor kinematics between wt and the lgc-46 RNAi strain. p = 0.015 Forward distance, 0.015 backward distance, 0.017 backward/forward ratio, 0.568 forward speed, and 0.844 backward speed. The asterisks indicate statistically significant differences compared with wt (*p < 0.05, **p < 0.01, ***p < 0.001) based on either one-way ANOVA with Tukey’s post hoc test (b–d) or unpaired two-sided t-test (f). The numbers inside brackets indicate sample size (n). n = numbers of independently recorded cells in b–d, but numbers of individual worms in f. Data are presented as mean values ± SEM. Source data are provided as a Source data file.