Extended Data Figure 5. oig-1 mutants defects and their rescue.

a: oig-1 mutants display locomotory defects. Locomotion of L4 animals was analyzed with tracking assays (Yemini et al 2013). The graphs on the left side of each panel correspond to assays comparing wild-type, oig-1 mutant, and oig-1; unc-30p::oig-1 animals. The graphs on the right side of each panel correspond to assays comparing wild-type, oig-1 mutant, and oig-1; oig-1^fosmid::gfp animals. Twenty animals (each dot on a plot) were tracked for each genotype for both comparisons. Mean and Q values are indicated. Note that in a previously published analysis of a large panel of available mutants, the same set of locomotory defects that we describe here for oig-1 mutants were found to be affected in unc-55 mutants ²⁸, albeit in a stronger manner than oig-1 mutants. Also note that the very strong locomotory defects unc-30 defects are qualitatively very different from oig-1 mutants, but this is to be expected since unc-30 mutant do not only show the synaptic defects that we describe here, but also lack the neurotransmitter GABA ³, thereby disabling any neuromuscular signaling.

i: The midbody speed of oig-1 mutant animals is significantly lower than that of wild-type animals. This defect is partially rescued (statistically different from oig-1 mutants but also from wild-type animals) by expressing unc-30p::oig-1 in oig-1 animals (left graph). The lower midbody speed of oig-1 mutants is completely rescued (statistically different from oig-1 mutants but not from wild-type animals) by expressing the oig-1^fosmid::gfp in oig-1 mutants.

ii: oig-1 mutants exhibit more dwelling than wild-type L4 animals. This defect is partially rescued (statistically different from oig-1 mutants but also from wild-type animals) by expressing unc-30p::oig-1 in oig-1 animals (left graph). The increased dwelling of oig-1 mutants is completely rescued (statistically different from oig-1 mutants but from not wild-type animals) by expressing the oig-1^fosmid::gfp in oig-1 mutants.

iii: oig-1 mutants exhibit an increased path curvature compared to wild-type animals. This defect is not rescued by expressing unc-30p::oig-1 in oig-1 animals (left graph). The increased path curvature of oig-1 mutants is completely rescued (statistically different from oig-1 mutants but not from wild-type animals) by expressing the oig-1^fosmid::gfp in oig-1 mutants.

b: Aldicarb-sensitivity defects in oig-1 mutants. oig-1 mutant young adult animals (red squares), which display aberrant GABAergic synapses in both the ventral and dorsal cord, show hypersensitivity to aldicarb-induced paralysis compared to wild-type (black triangles). Expression of oig-1^fosmid::gfp from a multicopy transgenic array (green circles) does not only rescue the oig-1 mutant phenotype, but even results in a slight hyposensitivity to aldicarb. Worms were tested every 15 minutes for paralysis by touching the head and tail three times each. n=20 for each strain, repeated 3 times.

c: Expression of oig-1 in the D-type neurons rescues ectopic DD synapses in the dorsal nerve cord. At the L1 stage when only the DD MNs are present, SNB-1::GFP (from juIs1-unc-25p::SNB-1::GFP) localizes to the ventral nerve cord (VNC) in wild-type animals (top left). Ectopic SNB-1::GFP puncta localize to the dorsal nerve cord (DNC) of oig-1 mutant L1s (top right). This phenotype is rescued by expressing oig-1 in the D-type MNs (unc-30p::oig-1, otEx4955, bottom left), but not by expressing oig-1 in the neighboring cholinergic MNs (unc-3p::oig-1, otEx4942, bottom right). White boxes indicate the dorsal nerve cord.