Picture from Culetto E et al. (2004) J Biol Chem "The Caenorhabditis elegans unc-63 gene encodes a levamisole-sensitive ...."

Expression pattern of the LEV-1::GFP construct. G, the lev-1::GFP construct was injected into the lev-1 mutant (allele e211). GFP expression is observed from embryo to the adult stage, where signal is found in a subset of neurons in the ventral nerve cord, indicated by arrows. H, LEV-1::GFP expression in body wall muscles. Scale bar = 50 μm.

Picture from Culetto E et al. (2004) J Biol Chem "The Caenorhabditis elegans unc-63 gene encodes a levamisole-sensitive ...."

Figure 4. Expression pattern of the UNC-63::GFP and the LEV-1::GFP constructs. A, structure of the injected GFP constructs. B, composite image of L3/L4 transgenic larvae stably transformed with UNC-63::GFP. The GFP signal is observed in a large number of neurons in the lumbar ganglia (lg), pre-anal ganglia (pag), head ganglia (hg), and ventral nerve cord (arrows indicate motor neuron cell bodies). Scale bar = 50 μm. C, UNC-63::GFP signal is shown in a subset of neurons in the anal ganglia. No expression is observed either in the sphincter muscle cell or in the anal depressor muscle. Scale bar = 5 μm. D, dorsal view of a young adult animal. All the body wall muscle cells, visible as large trapezoidal cells (example indicated by arrow), express UNC-63::GFP. A dark strip separates the left and right quadrants. Scale bar = 50 μm. E, four neurons (see arrows) of the posterior lateral ganglion express UNC-63::GFP. Scale bar = 5 μm. F, the UNC-63::GFP signal is expressed in some of the vulval muscle cells indicated by arrows. Scale bar = 10 μm. G, the lev-1::GFP construct was injected into the lev-1 mutant (allele e211). GFP expression is observed from embryo to the adult stage, where signal is found in a subset of neurons in the ventral nerve cord, indicated by arrows. H, LEV-1::GFP expression in body wall muscles. Scale bar = 50 μm.

Picture from Sato K et al. (2009) Proc Natl Acad Sci U S A "Differential requirements for clathrin in receptor-mediated endocytosis and ...."

Figure 3. Clathrin is required for neuromuscular function. (A-B'') CHC is concentrated presynaptically but is excluded from the active zone. (A-A'') Live worms co-expressing mRFP-CHC (A, A'') and the synaptic vesicle protein GFP-SNB-1 (A', A'') in GABA neurons were examined by fluorescence microscopy. (B-B'') Transgenic animals expressing GFP-CHC (B, B'') in the GABA neurons were stained with anti-GFP and anti-SYD-2 antibodies. Note that anti-SYD-2 antibody labels active zones in all synapses of the nerve cord (B'), whereas GFP-CHC labels only synapses in GABA motor neurons (B). (Scale bars, 10 um.) (C-F) Thrashing assays. (C) Wild-type L4 hermaphrodites (n = 8) and chc-1(b1024) (n = 8) were incubated at 20C for 24 h and then transferred to M9 buffer. Head thrashes were counted. (D) Wild-type L4 hermaphrodites (n = 8) and chc-1(b1025ts) mutants (n = 8) were incubated at 15C for 24 h and examined. (E) Wild-type L4 hermaphrodites (n = 5), chc-1(b1025ts) (n = 5), and chc-1(b1025ts) mutant animals expressing GFP-CHC (dkEx6 [Prgef-1::GFP::chc-1]) under the control of a neuron-specific promoter (n = 5) were incubated at 25C for 24 h before examination. (F) Wild-type young adult worms (n = 4) or chc-1(b1025ts) mutants (n = 8) were incubated at 30C for 10 min and examined. (G) Aldicarb resistance. Thirty young adult hermaphrodites of the wild type, chc-1(b1025ts), and lev-1(e211) were placed on aldicarb plates (0.5 mM) at the semi-permissive temperature of 22C and checked for their ability to move at the indicated time-points in response to prodding. lev-1(e211) mutants that lack a subunit of the muscle inotropic ACh receptor were resistant to both aldicarb and levamisole and used as a control. (H) Levamisole sensitivity of chc-1(b1025ts). Thirty young adult hermaphrodites of the wild type, chc-1(b1025ts), and lev-1(e211) were placed on levamisole plates (60 uM) and assayed for movement in response to prodding.

Picture from Tu, Shasha et al. (2023) MicroPubl Biol "Characterizing Three Azides for Their Potential Use as C. elegans ...."

Figure 1. KN3 as an alternative anesthetic for C elegans-related experiments:(A) Levamisole (Lev) and 1-Phenoxy-2-propanol (1P2P, 10%) are ineffective at paralyzing worms in chemotaxis assays to IAA. 1Lev, 1 mM levamisole; 10Lev, 10 mM levamisole. (B) All tested azides do not evoke chemotactic behavior. (C) IAA chemotaxis doesn't change using KN3 as an anesthetic, while it changes significantly using either TMSA or DPPA as an anesthetic. (D) 2-NN chemo-avoidance doesn't change using KN3 as an anesthetic, while it changes significantly using either TMSA or DPPA as an anesthetic. (E) Time courses showing the recovery of the worms in 18 minutes after the chemotaxis assays, statistical analysis was performed by comparing the area under the curve. (F) NaN3, KN3, TMSA, and DPPA all showed a certain extent of paralyzing effect in the buffer-based assay. Data in (A)-(D) and (F) are presented as box plots with all data points in magenta; the box plot displays the median, interquartile range, and minimal and maximal values. Data in (E) is presented as a line graph showing the median and interquartile range for the data at the indicated time point. Numbers above the x-axis denote the sample scale of each group; if the number is colored blue, the data is not normally distributed. vs NaN3 in (A) and (C)-(F); vs control in (B); *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, not significant.

Picture from Simon DJ et al. (2008) Cell "The microRNA miR-1 regulates a MEF-2-dependent retrograde signal at ...."

Figure 1. miR-1 Affects Muscle Sensitivity to ACh. (A) A transcriptional reporter containing a 3.7 kb mir-1 promoter driving expression of GFP showed expression in body-wall and pharyngeal muscles.(B) Body-wall muscle expresses two classes of nAChR: the ACR-16/alpha 7 homo-pentamers and the levamisole sensitive hetero-pentamer containing UNC-29. The ACR-16 receptor is activated by ACh while the UNC-29 receptor is activated by both ACh and levamisole (Lev).(C) The time course of levamisole (0.2 mM) induced paralysis of WT and mir-1(gk276) was compared.(D and E) Levamisole (100 μM, 0.5 s) evoked currents in body muscles were compared in WT (n = 18), unc-29(x29) (n = 3), and mir-1(gk276) (n = 9). Averaged traces (D) and peak amplitudes (E) are shown.(F and G) Acetylcholine (500 μM, 0.5 s) evoked currents in body muscles were compared in unc-29 and mir-1 unc-29 mutants. Averaged traces (F) and peak amplitudes (G) are shown. (asterisk) indicates changes that are significantly different (p < 0.01, Mann-Whitney) from control strains.The mean endogenous EPSC amplitudes (H) and cumulative probability distributions of endogenous EPSC amplitudes (I) and decay taus (J) for mir-1 and WT controls are compared.(K) Average endogenous EPSCs are shown for WT (black), mir-1(red), and a scaled version of mir-1 (blue).The error bars in (C), (E), (G), and (H) indicate standard error of the mean (SEM).