Figure 8.

Zebrafish Deficient for RyroxD1 Show Sarcomeric Disorganization, Myofibrillar Aggregates, and a Defect in Swimming

(A) Quantitative PCR for ryroxd1 mRNA levels in control (uninjected) and Ryroxd1 morpholino-injected (Ryroxd1 ex2 splice MO) zebrafish at 2 dpf.

(B) Western blot for Ryroxd1 protein and α-tubulin loading control in control (uninjected) and Ryroxd1 morpholino-injected zebrafish (D = Ryroxd1 double morpholino, ATG = Ryroxd1 ATG morpholino, and splice = Ryroxd1 splice-site targeting morpholino) at 48 hpf demonstrates effective reduction of Ryroxd1.

(C) Ryroxd1 ATG-single and ATG/splice double morphants show a significant reduction, of 48% and 73% respectively, in maximum acceleration in a touch-evoke response assay at 2 dpf compared to control zebrafish injected with a GFP targeting morpholino (Cont).

(D and E) Antibody labeling of Ryroxd1 double morphants at 48 hpf (D) and 96 hpf (E) for Actinin2 and phalloidin show disruption of muscle structure compared to uninjected controls.

(E) At 96 hpf, Ryroxd1 double morphants show severe disruption of the musculature with remnants of fragmented muscle fibers evident (arrows).

For (A) and (C), error bars represent SEM for three independent replicate experiments comprising 15 fish in each, ^∗∗p < 0.01.