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. 2022 Jul 12;119(29):e2113545119. doi: 10.1073/pnas.2113545119

Fig. 1.

Fig. 1.

Levamisole induces different behavioral responses in wild type and lev-10 but elicits same current on muscle cells. (A) Schematic of L-AChR clustering machinery. MADD-4/Ce-Punctin is secreted by motor neurons and localizes the heteromeric AChRs composed of UNC-38, UNC-63, LEV-1, LEV-8, and UNC-29. LEV-10, OIG-4, and LEV-9 are part of the clustering machinery. MOLO-1 is an L-AChR auxiliary subunit. (B) Percentage of paralyzed animals at 4 h and after overnight exposure to levamisole 1 mM. Mean values ± SD were plotted (wild type, n = 6; unc-29, n = 8; lev-10, n = 8; lev-9, n = 6; and madd-4, n = 3, where n is the number of independent experiments; to total number of animals tested per genotype: 60 to 160). (C) Representative traces and mean amplitude of currents evoked by 0.1 mM pressure-ejected levamisole on muscle cells hold at −60 mV. In this figure and all the figures, bar represents median, square represents mean, boxes represent quartiles, and whiskers represent SD. Student t test P = 0.36 (wild type, n = 19; lev-10, n = 13). (D) Morphologic data for wild type, lev-10, and unc-29 before and during exposure to 1 mM levamisole (n = 8 for the 3 genotypes except at 120 min for wild type and unc-29, n = 7). The ratio was compared using 1-way ANOVA at 0, 3, and 120 min for wild type (P < 0.0001), lev-10 (P < 0.0001), and unc-29 (P = 0.07) followed by Holm-Sidak posttests: ns, nonsignificant, *P < 0.05. (E) Representative pictures of wild type and lev-10 mutants expressing GCaMP3 in body muscles before (0) or after 3 and 120 min-exposure to 1 mM levamisole (16-color lookup table). Scale bar is 0.1 mm. (F) Calcium imaging data for wild type, lev-10, and unc-29 mutants during levamisole exposure. Young adults expressing GCaMP3 in body muscle were imaged on regular NGM (Nematode Growth Medium) plates at t = 0 min and were then put on 1 mM levamisole plates. Different cohorts of animals have been imaged at 0, 1, 3, 10, 15, 30, 60, 120, and 240 min of levamisole exposure, n > 15 for each genotype at each time point except at 10 min for lev-10 and unc-29, n = 9 and n = 10, respectively (see also SI Appendix, Table S1). The calcium index was calculated as indicated in Material and Methods. Mean currents ± SD were plotted.