Fig. 3

Fig. 3

Gmppb knockdown(KD) in Zebrafish causes abnormal development of muscle and motor neurons, movement disability. a Temporal expression of zebrafish gmppb from fertilization to day 3, and gapdh was used as control. b WISH shows the spatial expression pattern of gmppb using a dig-labeled antisense probe. gmppb was ubiquitously expressed in zebrafish embryos from 50%-epi to 24hpf. Scale bar: 250 μm. c Immunoblot of entire zebrafish tissue extracts indicates that gmppb MO injection effectively decreased Gmppb protein level. MO injection was performed at the one cell stage. Actin was used as a loading control. d Expression of myod in zebra fish, with or without gmppb MO injection, determined by WISH. The black rectangles label the position of enlarged views (bottom). Scale bar: 250 μm. e Gmppb KD results in decreased HuC (green) expression. Tg [HuC: GFP] transgenic zebrafish was injected with control MO (NC), or gmppb MO alone or together with mRNA encoding GMPPB. HuC expression was observed at 48 hpf. Scale bar: 100 μmfgmppb mutant larvae display decreased motor ability. Representative images and summarized movement path of zebrafish injected with control MO (NC) or gmppb MO. (Upper). Bar graph summarizes the moving distance and moving speed of zebrafish injected with control MO (NC) or gmppb MO (Lower). Motor ability was monitored in 2 dpf larvae. g. GMPPB is required for motor neuronal development in zebra fish. Tg [hb9: GFP]^ml2 transgenic zebrafish was injected with control MO (NC), or gmppb MO alone or together with mRNA encoding GMPPB at one-cell stage. Top: morphology of CaP axons was observed at 24 and 48hpf; bottom: statistical results of the length of CaP axons. Scale bar: 100 μm. Mean ± SD, ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05. p values were calculated using one-way ANOVA, Tukey’s multiple comparisons test