Figure 9. GLR glia-defective animals display locomotion abnormalities and hypersensitivity at high-salt concentrations.

(A–C) Images of L4 animals showing expression of gly-18prom::gfp reporter in (A) wild-type, (B) GLR glia ablation and (C) GLR glia ablation; ced-3(n717) backgrounds. GLR glia are outlined by red dashed circles. Black asterisks denote expression in a neuronal cell just above the dorsal GLR glia and red arrowheads point to HMC cells. In the GLR glia-ablation background, the nerve ring is anteriorly displaced as noted by a head muscle arm (gray arrowhead) penetrating the nerve ring at the anterior pharynx bulb. The HMC sister cell that normally dies by programmed apoptotic cell death in a wild-type animal, survives and also expresses gly-18prom::gfp in the ced-3(n7171) mutant background in (C). (D–K) Locomotion parameters of foraging wild-type (black), control (gray), GLR glia-defective (red), and GLR glia-ablated (purple) animals were analyzed using automated tracking (Katz et al, 2018; Katz et al, 2019). (D) change of direction frequency, (E) “front to back” reversal frequency, (F) % time spent paused, (G) average speed, (H) omega turn frequency, (I)% direct “back to front” reversal after backward movement, (J) % time spent moving backwards, (K) mean duration of backward movement. Black lines in dot plots indicate averages. (L) GLR glia-ablated animals paralyze at a significantly higher rate than wild-type animals and animals in which LET-381 is downregulated [let-381(ns1026) and LET-381 AID knockdown] when exposed to 300 mM NaCl. GLR glia-ablated animals also recover motility at a significantly lower rate. Data information: unpaired t test was used for statistical analysis in (D–L), *<0.05 in (L). For (L), 4 replicates were performed per genotype, n = 20–25 for replicate, error bars indicate standard error of the mean. Anterior is left, dorsal is up, and scale bars are 10 μm for all animal images. Source data are available online for this figure.