ZFIN ID: ZDB-FIG-220131-219
Liu et al., 2021 - GMPPB-congenital disorders of glycosylation associate with decreased enzymatic activity of GMPPB. Molecular biomedicine   2:13 Full text @ Mol Biomed
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 1-cell to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Long-pec

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

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP knu3Tg control Long-pec nervous system IFL
knu3Tg + MO1-gmppb standard conditions Long-pec nervous system IFL
gmppb AB standard conditions 1-cell whole organism RTPCR
1k-cell whole organism RTPCR
50%-epiboly whole organism ISH
50%-epiboly whole organism RTPCR
Bud whole organism ISH
Bud whole organism RTPCR
14-19 somites whole organism ISH
14-19 somites whole organism RTPCR
Prim-5 whole organism RTPCR
Prim-5 whole organism ISH
Long-pec whole organism RTPCR
Protruding-mouth whole organism RTPCR
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-gmppb WB AB control Prim-5 whole organism
WB AB + MO1-gmppb standard conditions Prim-5 whole organism
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-gmppb standard conditions Prim-5 whole organism Ab1-gmppb labeling decreased amount, abnormal
Long-pec multicellular organismal locomotion decreased process quality, abnormal
knu3Tg + MO1-gmppb standard conditions Long-pec nervous system EGFP expression decreased amount, abnormal
ml2Tg + MO1-gmppb standard conditions Prim-5 CaP motoneuron axon decreased length, abnormal
Long-pec CaP motoneuron axon decreased length, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Mol Biomed