ZFIN ID: ZDB-FIG-140902-102
Pinzón-Olejua et al., 2014 - Essential roles of zebrafish rtn4/Nogo paralogues in embryonic development. Neural Development   9:8 Full text @ Neural Dev.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Day 5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Day 5

Fig. 5

Development of neural and non-neuronal structures in rtn4b-knockdown, GFP transgenic embryos. At 1 dpf, tg(Shh:GFP), rtn4b morphant embryos showed deformations in the hypothalamic region (h), dorsal thalamic structures (arrow), anterior floor plate (f) and forebrain (A) and (B). Additionally, the notochord (n) is abnormally undulated (compare (C) and (D)). In addition, 1-dpf tg(hb9:GFP), rtn4b morphant embryos had fewer cell bodies in the spinal cord and truncated motor axons (E) and (F) (arrowheads). At 2 dpf, motor axons showed aberrant branching (G) and (H) (arrows). At 5 dpf, lateral (J) and dorsal (L) views reveal smaller eyes (e), aberrant retinal ganglion cells (RGCs) (white arrow) and few retinal axons projecting into the tectum (t), which is displaced anteriorly, along with abnormally patterned branchial arches (br) and reduced pectoral fins (pf) (I) to (L). At 2 dpf, tg(Isl1:gfp), rtn4b-MO1 morphants showed severe defects in cranial motor neurons, including lack of pairs III and IV (M) and (N). (O) Quantification of the number of motor neurons in posterior segments (n = 15 to 20) of the spinal cord in control morpholino (Co MO; n = 20), rtn4b-MO1 (n = 25) and rescued (n = 20) embryos at 1 dpf. (P) Proportion of embryos with abnormal motor axons in embryos injected with Co MO, rtn4b-MO, rtn4b-MO2, or coinjected with half concentrations of both MOs, or rtn4b-MO1 + rtn4b-mRNA, and injected with rtn4b-mRNA. hb: hindbrain; e: eye; ob: olfactory bulb; c: cerebellum; X: branchiomeric nerve X. Co MO (n = 15), rtn4b-MO1 (2.5 ng) (n = 30), rtn4b-MO2 (2.5 ng) (n = 31), rtn4b-MO1 (5.0 ng) (n = 17), rtn4b-MO2 (5.0 ng) (n = 11), rtn4b-MO1 + rtn4b-MO2 (2.5 ng) (n = 20), rtn4b-MO1 (5.0 ng) + rtn4b-mRNA (100 pg) (n = 69) and rtn4b mRNA (100 pg) (n = 85). Scale bar = 100 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP ml2Tg control Prim-5 motor neuron IFL
Prim-5 motor neuron axon IFL
Long-pec motor neuron IFL
Long-pec motor neuron axon IFL
ml2Tg + MO2-rtn4b standard conditions Prim-5 motor neuron IFL
Prim-5 motor neuron axon IFL
Long-pec motor neuron IFL
Long-pec motor neuron axon IFL
rw0Tg control Long-pec cranial motor neuron IFL
Long-pec cranial nerve III IFL
Long-pec cranial nerve IV IFL
Long-pec cranial nerve X IFL
rw0Tg + MO2-rtn4b standard conditions Long-pec cranial motor neuron IFL
Long-pec cranial nerve X IFL
t10Tg control Prim-5 floor plate IFL
Prim-5 hypothalamus IFL
Prim-5 notochord IFL
Day 5 cerebellum IFL
Day 5 eye IFL
Day 5 olfactory bulb IFL
Day 5 optic tectum IFL
Day 5 pharyngeal arch 3-7 IFL
t10Tg + MO2-rtn4b standard conditions Prim-5 floor plate IFL
Prim-5 hypothalamus IFL
Prim-5 notochord IFL
Day 5 cerebellum IFL
Day 5 eye IFL
Day 5 olfactory bulb IFL
Day 5 optic tectum IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ml2Tg + MO2-rtn4b standard conditions Prim-5 motor neuron axon guidance process quality, abnormal
Prim-5 spinal cord has fewer parts of type motor neuron, abnormal
Long-pec motor neuron axon guidance process quality, abnormal
Long-pec spinal cord has fewer parts of type motor neuron, abnormal
rw0Tg + MO2-rtn4b standard conditions Long-pec cranial nerve III absent, abnormal
Long-pec cranial nerve IV absent, abnormal
t10Tg + MO2-rtn4b standard conditions Prim-5 dorsal thalamus deformed, abnormal
Prim-5 floor plate anterior region deformed, abnormal
Prim-5 forebrain deformed, abnormal
Prim-5 hypothalamus deformed, abnormal
Prim-5 notochord undulate, abnormal
Day 5 cranial nerve II immature, abnormal
Day 5 eye decreased size, abnormal
Day 5 optic tectum mislocalised anteriorly, abnormal
Day 5 pectoral fin aplastic/hypoplastic, abnormal
Day 5 pharyngeal arch 3-7 absent, abnormal
Day 5 retinal ganglion cell immature, abnormal
Day 5 retinal ganglion cell neuron projection immature, abnormal
Day 5 ventral mandibular arch absent, abnormal
Acknowledgments:
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