Picture from Chitturi J et al. (2018) PLoS Genet "The UBR-1 ubiquitin ligase regulates glutamate metabolism to generate ...."

Fig 4. Removing GOT-1 transaminase restores bending in ubr-1 mutants. A) A diagram depicting the structure of GOT-1 and the molecularlesion of the hp731 allele. B) GOT-1 is ubiquitously expressed in all somatic tissues. Scale bar 10um. C) Mutations in got-1 suppress motor neurondefects in ubr-1. D) Restoring GOT-1 in premotor INs including AVE and RIM (blue line) in ubr-1; got-1 reverted curvature to that of ubr-1. P<0.05, P<0.01, P<0.001 by the Two-way RM ANOVA test. Data are represented as mean +- SEM.

Picture from Chitturi J et al. (2018) PLoS Genet "The UBR-1 ubiquitin ligase regulates glutamate metabolism to generate ...."

Fig 2. A critical requirement of UBR-1 in premotor interneurons during reversals. A) The UBR-1::GFP reporter was expressed in a subset of neurons including thepremotor interneurons of the reversal circuit (Premotor INs), the pharyngeal muscles, the body wall muscles, and hypodermal seam cells. Scale bar 10um. B) Reducedbending in ubr-1 mutants (grey line) was rescued by restoring UBR-1 expression in neurons (red line), but not in muscles (blue line). C) Reduced bending in ubr-1mutants (grey line) was robustly rescued by restoring UBR-1 expression in glutamate-receptor-expressing, premotor interneurons (INs), not in cholinergic or GABAergicmotor neurons (MNs). D) Among the premotor interneurons (INs), a robust rescue required UBR-1 expression in AVE and RIM (red line). P<0.05, P<0.01, P<0.001 by Two-way RM ANOVA. Data are represented as mean +- SEM.

Picture from Gupta N et al. (2024) MicroPubl Biol "A DTC morphometrics package for quantification of complex and variable cellular ...."

Figure 1. Overview of image processing and markup.: A) A maximum projection image of an Z-confocal series (file format .nd2), with no contrast adjustment or filtering. B) Maximum projection image created from the same image file, using "Max project and change contrast" command. C) Same panel as in B, with yellow segmented line showing path used in "Rotate and straighten based on curved line" command. The arrow indicates the direction the line must be drawn to ensure the orientation is correct (distal to the left) when rotated. D) Final rotated, straightened, cropped image. E) Cropped image with the large circle positioned tangent to the curve of the distal end of the gonad and the smaller circle placed around the center of the nucleus. Both circles are placed manually by user. F) Same image as in E, with features manually curated by user. G) Screenshot of spreadsheet output that results from "Measure all markups" command. H) Binary image that results from "Prepare for Sholl plugin" command. Note that for visual clarity, the contrast in this image has been inverted from the actual result in ImageJ. I) Same image as in H, with Sholl radii shown every 1µm. J) Screenshot of spreadsheet output from the Neuroanatomy plugin that runs the Sholl analysis (Ferreira et al. 2014). The screenshot has been trimmed to show the number of intersections that occur in the first 7 and final 3 radii. Worms imaged bear the qIs57[lag-2p::GFP] transgene (Seigfried et al., 2004; Davis et al., 2022). Scale bar is 10 µm in all images.

Picture from Robinson, Bryan et al. (2019) MicroPubl Biol

Figure 1 The RIS interneuron promotes head movement quiescence but not locomotor quiescence during lethargus and stress-induced sleep. (A-D) Locomotor rate assessed by body bends per minute among animals during the L4-to-adult lethargus (A), during the young adult stage (B), and during stress-induced sleep at the young adult stage following exposure to ultraviolet light (C) or Cry5B pore-forming toxin (D). Animals were examined on thin even lawns of E. coli OP50 and examined for the number of sinusoidal body bends (peak-to-peak movement of any body part) in one minute. To select mid-lethargus animals, late-L4 larvae were identified based on vulval morphology, screened one hour later for cessation of pharyngeal pumping as an indicator of lethargus entry, and scored 30 min later for bend rates. For UV-SIS, worm plates were placed lid-down on a 302nm 60mW UV light box for 50 sec and examined 60 min later. For Cry5B-SIS, animals were exposed to Cry5B-expressing bacteria for 5 min, transferred to a thin even lawn of OP50, and examined 10-15 min later. For all bend rate analyses, plates were gently moved to the stereomicroscope field of view and allowed to sit for 1 min prior to examination. Each animal was examined for 1 min and a body bend was defined as a peak-to-peak unit of sinusoidal motion. **P<0.0001 vs. N2, one-way ANOVA with Dunnett's multiple comparisons test. N=15-20, experimenter blind to genotype. (E-L) Categorization of movement during SIS following UV exposure (E-H) and during exposure to Cry5B (I-L). Each animal was examined for 5 sec and characterized as completely immobile (E and I), head mobile but body immobile (F and J), or head mobile plus body movement of at least 1/10 body-length (G and K). Two-way repeated measures (RM) ANOVA with time as the repeated factor and Sidak's multiple comparisons test reveals that more ceh-17(lf) than aptf-1(lf) animals are completely immobile under certain SIS conditions (Panel I, ceh-17 vs. each aptf-1 mutant, P<0.05 at 40 min). However, aptf-1 mutants are impaired for head movement quiescence (N2 vs. aptf-1 mutants P<0.05 at multiple time points during UV-SIS and Cry5B-SIS), and not significantly impaired for body movement with the exception of the 30 min time point of UV-SIS, for which aptf-1(tm3287) is different from wild type N2 (P<0.05). H and L show stacked categories of behavior for the time points indicated by shaded boxes within the time courses. Three trials of 25 animals were performed and the experimenter was blind to genotype.

Picture from Browning H et al. (1996) Development "A sperm-supplied factor required for embryogenesis in C. elegans."

Figure 6. Immunofluorescence analyses of SPE-11 localization during spermatogenesis. In each confocal micrograph (A,C,D) spermatogenesis proceeds from left to right. SPE-11 is stained with affinity purified antiserum to SPE-11, DNA is stained with propidium iodide or 4, 6 diamidino-2-phenylindole (DAPI), andnuclear envelopes are stained with a monoclonal antibody (mAb414) to nuclear pore complex (NPC) proteins (Davis and Blobel, 1986). Scale bar, 10 µm. (A) Confocal images of a wild-type male gonad double labeled with antiserum to SPE-11 and propidium iodide. Each micrograph is the projection of a Z series containing seven sequential focal planes including approximately half the gonad thickness. SPE-11 first appears as dots in the nuclei of primary spermatocytes. The dots appear to coalesce before the first meiotic division. During the first and second meiotic divisions, SPE-11 is diffuse throughout the cells (bracket). SPE-11 is localized in a ring around the nuclei of sperm. (B) Conventional epifluorescence micrographs of sperm double labeled with antiserum to SPE-11 and DAPI. SPE-11 is perinuclear in sperm. Inset: Four spermatids attached to a single residual body. Some staining is detectable in the residual body. (C) Confocal images of primary spermatocytes double labeled with antiserum to SPE-11 and propidium iodide. The micrographs are projections of a Z series that includes the entire depth of the tissue. As the DNA becomes condensed in preparation for the first meiotic division, SPE-11 forms a fenestrated ring around the DNA. As discrete chromosomes become visible, SPE-11 relocalizes throughout the nucleus and is sometimes localized between the chromosomes. (This is also evident from analysis of individual focal planes; data not shown.) (D) Confocal images of primary spermatocytes double labeled with antiserum to SPE-11 and NPC antibodies. The micrographs are projections of a Z series that include the entire depth of the tissue. A merge of the NPC and SPE-11 projections demonstrates that the dynamic relocalization that SPE-11 undergoes in primary spermatocytes occurs within the nucleus. Spermatogenesis proceeds from left to right, with the exception of a few sperm (small rings stained with antiserum to SPE-11) in the upper left portion of the micrographs. There is no detectable staining of these sperm with the NPC antibody.