<i>fshr-1</i> re-expression in cholinergic and GABAergic neurons is sufficient to restore neuromuscular function but exacerbates synaptic vesicle accumulation defects.

<p>Behavioral (A, C) and synaptic structure (B, D) effects of genomic <i>fshr-1</i> DNA re-expression in the cholinergic (ACh) neurons (A, B) or GABAergic neurons (C, D) of <i>fshr-1(ok778)</i> mutant animals compared to wild type and <i>fshr-1(ok778)</i> worms. (A, C) (<i>Upper panels</i>) Representative (<i>left</i>, n = 3 plates/strain) and cumulative pooled (<i>right</i>) aldicarb data ± s.e.m. showing complete and even hyper-rescue of aldicarb paralysis in worms with cholinergic neuron-specific <i>fshr-1</i> rescue (ACh Neuron Rescue) (A) and nearly complete rescue of wild type paralysis in worms with GABAergic neuron-specific rescue (GABA Neuron Rescue) (B). Scatter points show individual plate averages. (<i>Lower panels</i>) Box and whisker plots showing mean body bends per minute from swimming assays performed on n = 30 young adult animals of each genotype. Minima and maxima (whiskers) are shown, as well as the first and third quartiles of data (boxes), divided by the median line. The “X” denotes the mean value of the data set, and circles show individual data points. Note that while the GABA rescue worms have body bending rates that are partially restored to wild type levels as seen in the swimming assay, ACh rescue worms retain the reduced body bending rates seen with <i>fshr-1</i> mutants, likely due to the excessive muscle excitation caused by <i>fshr-1</i> re-expression to above wild type levels (see <i>Upper panels in</i> A vs. C). (B, D) Wild type worms, <i>fshr-1</i>(<i>ok778</i>) mutants, and ACh neuron rescue (B) or GABA neuron rescue (D) animals that also expressed GFP::SNB-1 in cholinergic (ACh) neurons were imaged using a 100x objective. (<i>Left panels</i>) Representative images of the dorsal nerve cords halfway between the vulva and the tail of young adult animals. (<i>Right panels</i>) Quantification of puncta (synaptic) intensity and puncta density (per 10 <i>μ</i>m) ± s.e.m. Puncta intensity is shown normalized to wild type. For (B), n = 29 animals imaged for wild type, n = 26 for <i>fshr-1</i>, and n = 32 for ACh Neuron rescue. For (D), n = 21 for wild type, 15 for <i>fshr-1</i>, and n = 24 for GABA Neuron rescue. For all data, statistical significance was analyzed using a one-way ANOVA and Tukey’s post hoc test or a Wilcoxon Rank Sum test followed by a Steel-Dwass multiple comparison analysis, as appropriate. Results of analyses for which <i>p</i> ≤ 0.05 are indicated by horizontal lines above the bars. <i>*p</i> ≤ 0.05, **<i>p</i> ≤ 0.01, <i>***p</i> ≤ 0.001, <i>****p</i> ≤ 0.001.</p> <p>(PDF)</p>