Picture from Hrus A et al. (2007) PLoS One "C. elegans agrin is expressed in pharynx, IL1 neurons and distal tip cells and does ...."

Agr-1::reporter expression in transgenic animals. B Expression starts in 2 cells in the anterior part of the embryo at around the 64 AB cell stage. C, Towards the end of gastrulation expression is seen in about 40 cells throughout the embryo including neuronal precursors, several ventral hypodermal cells and pharyngeal precursor cells (ventral view). D At the 1 1/2 to 2 fold stage expression is seen in IL1 neurons (identity determined postembryonically), embryonic motoneurons and a number of additional cells in the head, most likely arcade cells and epithelial buccal cells in the pharynx, and in few apoptotic cells (marked by +). E, In the 3fold stage embryos expression is seen in the IL1 neurons (6 neurons), most of the arcade cells (3 anterior arcade cells and the DL and DR posterior arcade cells) and the buccal epithelial cells in the pharynx. The 2 lateral IL1 neurons express GFP only weakly and only in early larval stages, wheras the remaining 4 IL1 neurons express GFP strongly throughout all larval stages. F (dorsal view) and I In L1 larvae expression is observed, in the buccal epithelial cells (dashed arrow), in 3 anterior arcade cells and the DL and DR posterior arcade cells (arrowheads), and in IL1v and IL1d neurons (arrows) and posterior gut cells (asterisk). In F, the worm was co-stained with DiI. G and H Head of a young adult worm; expression is visible in the buccal epithelial cells (dashed arrows) and in the IL1v and IL1d neurons (arrows); open arrowheads point at the IL1 processes in the nerve ring. J, L2 larva; expression in the migrating distal tip cells (arrows) and posterior gut (asterisk). Bars are 10um.

Picture from Hrus A et al. (2007) PLoS One "C. elegans agrin is expressed in pharynx, IL1 neurons and distal tip cells and does ...."

Figure 5. Agr-1::reporter expression in transgenic animals. A, Reporter genes were fused to different portions of agrin non-coding and coding sequences as shown in the schematic representation of the genomic region containing the agr-1 promoter and agr-1 5'-end. The lengths of the promoter or gene sequences and the names of the the pagr-1::reporter plasmids and DNA arrays are indicated. Since all of these constructs resulted in the same expression patterns, representative micrographs of the kdIs66 transgenic worms are shown in B-J. B Expression starts in 2 cells in the anterior part of the embryo at around the 64 AB cell stage. C, Towards the end of gastrulation expression is seen in about 40 cells throughout the embryo including neuronal precursors, several ventral hypodermal cells and pharyngeal precursor cells (ventral view). D At the 1 1/2 to 2 fold stage expression is seen in IL1 neurons (identity determined postembryonically), embryonic motoneurons and a number of additional cells in the head, most likely arcade cells and epithelial buccal cells in the pharynx, and in few apoptotic cells (marked by +). E, In the 3fold stage embryos expression is seen in the IL1 neurons (6 neurons), most of the arcade cells (3 anterior arcade cells and the DL and DR posterior arcade cells) and the buccal epithelial cells in the pharynx. The 2 lateral IL1 neurons express GFP only weakly and only in early larval stages, wheras the remaining 4 IL1 neurons express GFP strongly throughout all larval stages. F (dorsal view) and I In L1 larvae expression is observed, in the buccal epithelial cells (dashed arrow), in 3 anterior arcade cells and the DL and DR posterior arcade cells (arrowheads), and in IL1v and IL1d neurons (arrows) and posterior gut cells (asterisk). In F, the worm was co-stained with DiI. G and H Head of a young adult worm; expression is visible in the buccal epithelial cells (dashed arrows) and in the IL1v and IL1d neurons (arrows); open arrowheads point at the IL1 processes in the nerve ring. J, L2 larva; expression in the migrating distal tip cells (arrows) and posterior gut (asterisk). Bars are 10um.

Picture from Oswald, Mackenzi et al. (2020) MicroPubl Biol

Figure 1. Mutations in E3 ubiquitin ligase genes and glr-1 affect spontaneous reversal frequencies: A. Spontaneous reversal frequency assays were performed using C. elegans strains harboring mutations in E3 ubiquitin ligases. Box and whisker plots show the average reversals per minute, bounded by quartiles; the line in each box represents the median of the average reversals per minute for each genotype. N=23 individual animals measured for WT, hrd-1, and hrdl-1; N=24 individual animals for marc-6. * = p <0.05 (p=0.00069 for marc-6; p=0.032 for hrd-1); for hrdl-1, p= 0.14. Significance was calculated using the Tukey-Kramer test following a one-way ANOVA. The data is normally distributed (Shapiro-Wilk test, p=0.37) and groups show equal variance (residuals vs. fitted plot). B. Spontaneous reversal frequency for wild-type and glr-1 (n2461) animals. N=20 individual animals for both genotypes; significance was calculated using Student's t-test (p=0.0063). The data is normally distributed (Shapiro-Wilk test, p=0.52) and groups show equal variance (residuals vs. fitted plot). C. Schematics of E3 ubiquitin ligase genes and glr-1 (Oswald et al., 2020). Schematics were made and annotated using http://wormweb.org/exonintron. The mutant E3 ubiquitin ligase strain hrd-1 contains an indel, hrdl-1 contains a deletion and marc-6 contains a nonsense mutation resulting in a premature stop codon.

Picture from Suzuki M et al. (2004) Biol Chem "Metalloproteases with EGF, CUB, and thrombospondin-1 domains function in ...."

Figure 2. Expression pattern of nas-37p::gfp. DNA fragment extending from y3679 to q5 relative to the initiation ATG codon of the nas-37 gene was amplified from the genomic DNA extracted from C. elegans N2 animals with the primer 5'-GCG CAA GCT TTC ACC CAA AAA TTA TCA AGG - 3' and 5'-GGC CGT CGA CTT CAT TCT GCA AAA TAG AAC-3'. The fragment was digested with HindIII and SalI, and ligated into the promoterless vector pPD95.67, allowing in-frame fusion of the nas-37 sequence and the GFP reporter gene. Transgenic strains were generated by microinjecting the reporter plasmid (50 ug/ul) together with the marker plasmid pRF-4 rol-6 (su1006) as described previously by Kramer et al. (1990) (50 ug/ ul) into the syncytial gonad of wild-type nematodes (Mello et al., 1991). Panels (A) and (B) show L3 larvae. The large and small arrows in the panel (A) indicate the hypodermal and rectal grand cells, respectively. Panel (C) shows a young adult hermaphrodite. The arrow indicates the position of vulva. Panel (D) shows an adult hermaphrodite. The left is a confocal image and the right is the merge with its bright field image. The arrow indicates the vulva.

Picture from Voia, Anna et al. (2022) MicroPubl Biol

Figure 1. Monitoring of oxidative response in live C. elegans animals.(A) Schematic representation of C. elegans showing the regions of the animals that were imaged. (B) Epifluorescence images of wild-type (N2) L4 larvae treated with dihydroethidium. (C-D) Fluorescence levels of dihydroethidium measured in the pharynx of wild-type animals (N2; C) and isp-1(qm150) mutants (D) grown in presence of the compounds indicated on the horizontal axis. (E) Epifluorescence images of adult animals expressing DAF-16::GFP. (F-G) Nuclear:total cell (Nu:Cell) fluorescence ratio of DAF-16::GFP (F) and SKN-1B/C::GFP (G) measured in intestinal cells of adult animals grown in presence of the compounds indicated on the horizontal axis. (H) Epifluorescence images of adult animals expressing SKN- 1B/C::GFP. (I) Total fluorescence levels of SKN-1B/C::GFP measured in the intestine of adult animals grown in presence of the compounds indicated on the horizontal axis. In panels B, E and H, the schematic representation at the top left outlines the region where fluorescence levels were measured, and the images are taken from animals grown in control (top right), paraquat (bottom left) and paraquat+lupeol (bottom right) conditions. For all images, scale bar = 50 μm. For all graphs, black lines represent mean ± standard deviation, and statistical analyses were done using a one-way ANOVA test with a Tukey-Kramer post hoc test (*** = p < 0.001, ** = p < 0.01, ns = p > 0.05). F.I. = fluorescence intensity, a.u. = arbitrary units.

Picture from Hu Z et al. (2011) Neuron "A neuropeptide-mediated stretch response links muscle contraction to changes ...."

Figure 3. The NLP-12 Receptor CKR-2 Is Also Required for Suppression of Aldicarb-Induced Potentiation (A, B, D, and E) Endogenous EPSCs (A and B) and stimulus-evoked EPSCs (D and E) were recorded from body wall muscle of adult worms of the indicated genotypes, with (blue) and without (black) aldicarb pretreatment (30 min for ACh rescue, 60 min for all others). Representative traces of endogenous EPSCs (A), averaged stimulus-evoked responses (D), and summary data (B and E) are shown. ckr-2 mutants containing transgenes driving CKR-2 expression in all cholinergic neurons (using the unc-17 promoter) [rescue (ACh)], in GABA neurons (using the unc-25 promoter) [rescue (GABA)], and in cholinergic motor neurons (using the acr-2 promoter) [rescue Pacr-2] are indicated. (C) Aldicarb-induced paralysis is compared for the indicated genotypes. The number of animals analyzed (B and E) or the number of replicate experiments (C) is indicated for each genotype. (F) Pckr-2::NLS::GFP expresses in both cholinergic neurons (Punc-17::mCherry), in GABA neurons (Punc-25::mCherry). Arrows indicated cells bodies. Values that are significantly different from untreated wild-type controls are indicated (***p < 0.001;*p < 0.05). Error bars indicate SEM. A common set of wild recordings (+ and - aldicarb) were utilized in Figures 1, 2, 3, and 5. All comparisons to this control data set were corrected for multiple comparisons using the Tukey-Kramer test. All electrophysiological data are shown in Table S3.

Picture from Oswald , Mackenzi et al. (2020) MicroPubl Biol

Figure 1. Mutations in E2 ubiquitin-conjugating enzyme genes and glr-1 affect spontaneous reversal frequencies differently: A. Spontaneous reversal frequency assays were performed using wild type (WT) C. elegans, alongside strains harboring mutations in glr-1 or in genes that encode the E2 ubiquitin-conjugating enzymes, ubc-6 and ubc-7. Box and whisker plots show the average reversals per minute, bounded by quartiles; the line in each box represents the median of the average reversals per minute for each genotype. N=20 individual animals for all genotypes; significance relative to WT was calculated using the Tukey-Kramer test following a one-way ANOVA (*, p=0.0027 for glr-1; non-significant: p=0.58 for ubc-6; p=0.48 for ubc-7). The data is normally distributed (Shapiro-Wilk test, p=0.86) and groups show equal variance (residuals vs. fitted plot). B. Schematics of E2 ubiquitin-conjugating enzyme genes and glr-1 gene, including the locations of point mutations. Schematics were made using http://wormweb.org/exonintron. C. Crystal structures of the human E2 Ube2j2 (top, with rotated view to show the back side of the protein) and Ube2g2 (bottom, left). Models were made using Pymol. Residues that are mutated are shown in red (Ube2j2, D153; UBC-6, T158) and yellow (Ube2g2, P78, UBC-7, P79). Residue F156 of Ube2j2 is shown in cyan. Alignments of C. elegans UBC-6 and UBC-7 regions of homology, compared to putative orthologs in other eukaryotes. The highlighted residues correspond to those shown in the structures.

Picture from Zocher E et al. (2018) MicroPubl Biol "Dominant-negative VPS-4 disrupts ODR-10::GFP distribution but has limited ...."

Figure 1. Effects of expression of vps-4(E219Q) (VPS-4 DN) under the odr-10 promoter. A. Image of the head of an animal to illustrate the distribution of ODR-10::GFP fluorescence in an AWA neuron of an animal expressing VPS-4 DN under the odr-10 promoter. Animals were scored for having no visible fluorescence, fluorescence in the cilia, fluorescence in the cell body (arrowhead), with or without ciliary fluorescence (arrow). Red fluorescence is from the injection marker, Pttx-3::dsRed (asterisks). Image acquired at 630X magnification. B. Localization of ODR-10::GFP in the AWA neurons of adult hermaphrodites. Bar graph shows the relative distribution of phenotypes under different conditions. Numbers in the columns indicate the number of animals observed. Data are significant with p << 0.001 using a Chi-square contingency test, Chi-Square value = 123.82, 6 degrees of freedom. No assumptions were violated. C. Effects of different conditions on chemotaxis toward diacetyl, compared to ethanol control for young adult hermaphrodites; odr-10 (ky32) is shown as a negative control. Animals in chemotaxis experiments did not express ODR-10::GFP. Box and whisker plots show the results of 12-13 independent trials. Median chemotaxis indices were wild type (N2) = 0.85, wild type (N2), starved = 0.8, VPS-4 DN = 0.81, VPS-4 DN, starved = 0.7, odr-10 (ky32) = 0.08. Boxes bound the 1st and 3rd quartiles and circles represent outliers of greater than 1.5 times either quartile. Chemotaxis indices were calculated using the following expression: ((animals at chemoattractant)-(animals at control))/total animals on the plate. * p<0.05 compared to wild type, Tukey-Kramer post-test.

Picture from Hobson RJ et al. (2011) Curr Biol "Complexin maintains vesicles in the primed state in C. elegans."

Figure 1. cpx-1 Encodes the Worm Homolog of Complexin and Regulates Locomotion and Neurotransmitter Release (A) cpx-1 is the complexin gene with the highest identity to mouse and fly complexins. C. elegans CPX-1 was aligned with mouse CplxI (AAH14803) and fly complexin (AAF69518) using ClustalW2. Percent identities were calculated using Jalview [33]. (B) cpx-1 is expressed in the nervous system. Top: gene model and regions deleted in C. elegans complexin mutant alleles (red bars). The black line represents the region of the genome used to create the rescuing GFP construct. Gray triangle on transcript represents the 30UTR. GFP was inserted after the first three codons. Bottom: a reporter construct consisting of the cpx-1 gene with GFP inserted at the N terminus of complexin is expressed in many neurons in the head and body of the animal. Neurons expressing cpx-1 include the motor neurons of the ventral nerve cord, neurons in the head ganglia, and also neurons of the pharyngeal nervous system, including theMCand M4 motorneurons. The inset shows the punctate localization of GFP::CPX-1 along the dorsal nerve cord. Confocal images were taken of young adult hermaphrodites oriented dorsal up and anterior to the left. Images are a stitch of multiple confocal Z projections along the length of the animal. (C) cpx-1 mutants are defective for thrashing in liquid. cpx-1 thrashing behavior can be rescued by expressing cpx-1 under a panneuronal promoter (Prab-3::cpx-1 EG4633), but not by expressing it specifically in the acetylcholine neurons (Punc-17::cpx-1 EG4713). Graph shows means and standard error of the mean (SEM) of the thrashing behavior. Tukey-Kramer multiple comparisons test: *p < 0.001. The n values for each strain are listed at the bottom of each bar. (D) cpx-1mutants are hypersensitive to the acetylcholinesterase inhibitor aldicarb, indicating that excessive acetylcholine is released at neuromuscular junctions. This sensitivity can be rescued by expressing cpx-1 in all neurons using the rab- 3 promoter. Graph shows means and SEM.

Picture from Monteiro MI et al. (2012) Mol Biol Cell "The kinesin-3 family motor KLP-4 regulates anterograde trafficking of GLR-1 ...."

FIGURE 1. The kinesin KLP-4 functions in the VNC to regulate the abundance of GLR-1 and GLR-1-dependent behavior. (A) Protein domain organization of C. elegans KLP-4, Drosophila Khc-73 and mouse KIF13A and KIF13B. The N-terminal motor domain (red box) containing the signature ATP binding motif (thin black bar) and MT binding regions (thick black bar), the cargo binding domain (gray box), the Fork-head associated domain (FHA) (purple box) and the microtubule plus-end binding domain (CAP-Gly) (orange box) are shown. The percent identity of the motor domain of C. elegans KLP-4 (white percentages) or full length KLP-4 (black percentages) versus Drosophila Khc73, mouse KIF13A and KIF13B are indicated. (B-F) Representative images of the anterior VNC of larval stage 4 (L4) wild-type (B), klp-4(pz19) (C), rescued klp-4(pz19) (D), klp-4(tm2114) (E), and rescued klp-4(tm2114) (F) animals expressing an integrated GLR-1::GFP transgene (nuIs25). In these and all subsequent images anterior is to the left and ventral is up. (G-H) Quantification of GLR-1::GFP puncta intensities (normalized) (G) and densities (H) for the strains pictured in B-F. Means and SEM are shown for n = 38 wild type, n = 23 klp-4(pz19), n = 21 rescued klp-4(pz19), n = 20 klp-4(tm2114), and n = 21 rescued klp-4(tm2114) animals. (I) Quantification of number of spontaneous reversals per minute for n = 20 wild-type, n = 20 klp-4(tm2114), n = 23 animals expressing an integrated klp-4 transgene under the control of the glr-1 promoter (klp-4(xs)) (pzIs20), n = 14 glr-1(n2461), and n = 13 glr-1(n2461) animals overexpressing klp-4 (klp-4(xs)). Values that differ significantly from wild type are indicated by asterisks above each bar, whereas other comparisons are marked by brackets (**p 0.001, *p 0.01, #p 0.05, Tukey-Kramer Test).