<i>vt34</i> acts in a parallel pathway to reduce DAT-1 activity.

(A) Automated analysis of Swip reveals enhanced Swip relative to dat-1 in vt34; dat-1 double mutants. Single L4 animals were recorded using a video capture system and analyzed with SwimR software as described in Methods. Swimming frequency with SEM shaded above and below the mean is displayed. Data represent at least 20 animals per genotype analyzed by two-way ANOVA with multiple Bonferroni post-tests. Significant genotype x time interaction effects were evident with post-tests indicating P vt34, dat-1(ok157) vs. vt34; dat-1(ok157), and dat-1(ok157) vs. vt34; dat-1(ok157). (B) Over-expression of GFP::DAT-1 does not fully rescue vt34 Swip. Transgenic animal production and manual endpoint assays of swimming behavior using L4 animals were pursued as described in Methods. In these assays, 10 animals were assayed per well and at least 24 assays were performed per genotype. Data were analyzed using one-way ANOVA with multiple Bonferonni post-tests. *** = P vt34 animals expressing Pdat-1:GFP::DAT-1 exhibit normal DAT protein distribution as visualized in vivo using confocal microscopy. Single planes containing cell body (white arrow), synapse (open triangle) and dendrites (closed triangle) were imaged and quantified as described in Methods and Materials. Anterior is left, and represents ratiometric image of a CEP neuron. Scale bar represents 20 μm. (E) Quantification of GFP::DAT-1 fluorescence levels from images as in C and D, expressed as a ratio of synapse/cell body or dendrite/cell body in individual animals as described in Methods, analyzed by two-way ANOVA followed by Bonferroni post-tests. No significant genotype x region effect was observed nor were the individual compartments statistically different. Data derive from at at least 12 animals per genotype. Error bars represent SEM.