Figure 7. Molecules that disrupt coordinated invadopodia state and growth cone entry cause behavioral defects. A, Schematic of the behavioral assay. B, Representative collapsed image from 60-s movies of animals exposed to either 4°C or 23°C showing animals display a shivering movement after exposure to 4°C. C, Representative image of FLOTE segmentation used for the behavioral analysis. D, Method of calculating tail curvature. Curvature magnitude analyzed as the sum of angle A, angle between segments 2 and 3, and angle B, angle between segments 3 and 4. E, Representative graph of typical “turn/swim” bout. F, Two representative plots of spine curvature during shiver events. Note the scale on the y-axis demonstrates a shorter displacement then seen in turn bouts. G, Number of shivers quantified at 4°C (n = 24 at 2 dpf, 42 at 3 dpf, 42 at 4 dpf, and 48 at 5 dpf) and 23°C (n = 12 at 2 dpf, 21 at 3 dpf, 21 at 4 dpf, and 24 at 5 dpf). H, Total percentage of time shivering quantified at 4°C (n = 24 at 2 dpf, 42 at 3 dpf, 42 at 4 dpf, and 48 at 5 dpf) and 23°C (n = 12 at 2 dpf, 21 at 3 dpf, 21 at 4 dpf, and 24 at 5 dpf). I, Number of “turn/swim” bouts at 4°C (n = 24 at 2 dpf, 42 at 3 dpf, 42 at 4 dpf, and 48 at 5 dpf) and 23°C (n = 12 at 2 dpf, 21 at 3 dpf, 21 at 4 dpf, and 24 at 5 dpf). G–I, Cyan dots denote 4°C. Pink dots denote 23°C. J, Representative collapsed image from 60-s movies of animals exposed to 4°C and either treated with DMSO, 100 μm sp-cAMP, 25 μm rp-cAMP, or in a dcc+/− background treated with DMSO or 100 μm sp-cAMP. K, Total percentage of time shivering quantified at 4°C and 23°C (n = 4 sp-cAMP animals; 4 rp-cAMP animals; 5 dcc+/− animals; 3 dcc+/− +100 μm sp-cAMP animals). Cyan dots denote 4°C. Pink dots denote 23°C. L, Schematic representation of DCC-mediated invadopodia brake. Scale bars: 10 μm (B, J). G–I, K, Data are represented as mean ± SEM.