Figure 2

(A) Eight to 14 dpf pmm2^m/m embryos become progressively nonmotile. Graph shows number of nonmotile fish per day. Each colored dot represents the progeny from the same biological cross, such that all sibling animals collected on different days from that clutch are represented by the same-color dot. Genotyping showed all collected nonmotile embryos were pmm2^m/m. (B) Lateral images of Petri dishes show pmm2^m/m embryos lying on the bottom. Arrows indicate nonmotile pmm2^m/m embryos. (C) pmm2^m/m embryos die 9 to 14 dpf. Graph shows number of embryos dying each day. Each colored dot represents the progeny from the same biological cross, such that animals collected (on different days) from that clutch are siblings. (D) Schematic demonstrates Zebrabox behavioral analyses with 1 embryo placed per well and sensory deprived for 15 minutes. The swim paths, speeds, and general behavior are recorded for 10 minutes. Images of swim paths 6 to 12 dpf. Green paths indicate slow swim speed and red paths fast swim speeds. (E) Graphs show distance each embryo swam from 5–12 dpf at slow (upper) and fast (lower) swim speeds. n = >100 embryos per genotype over 5 experiments. Error bars show SEM, Student’s t test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (F) Graph illustrates number of swim events initiated per embryo 5–12 dpf. (G) Graph illustrates percentage of embryos actively swimming 4–14 dpf. (H) Confocal images of neuromuscular systems. Motor axons are stained green with acetylated tubulin, and AChRs are stained red with bungarotoxin. Lateral images show higher power images of boxed regions. Arrowheads indicate immature, disorganized postsynaptic density. Scale bars: 20 µm and 8 µm. (I) Graphs show area of bungarotoxin staining. Each dot represents the average area of 15 synapses in an individual embryo. n = 8 embryos per condition. Error bars show SEM, Student’s t test, ****P < 0.0001.