ZFIN ID: ZDB-FIG-140902-100
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:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec

Fig. 3

Rtn4a and Rtn4b protein levels and morpholino knockdown. (A) Western blot (WB) analysis using the zebrafish α-Rtn4a antibody 15] shows that Rtn4a-l (an approximately 43-kDa band, arrow) is suppressed either by injection of a mixture of morpholinos (MOs) against each rtn4a isoform or a MO against the rtn4a-l isoform. Embryos expressing an Rtn4a-l-GFP fusion can overcome the Rtn4a MO1 downregulation. GFP antibodies detect the fusion protein at approximately 70 kDa and GFP alone at approximately 26 kDa. α-tubulin (Tub) served as a protein loading control. (B) Exogenous Rtn4al-GFP was detected at 6 hpf, but simultaneous coinjection of rtn4a-l MO1 abrogated its expression. (C) α-Rtn4b antibodies show the downregulation of Rtn4b for both MOs used in the experiments. The 180-kDa band (arrowhead), probably a dimerization band, is entirely reduced in MO-injected embryos, and the 90-kDa band (arrow) shows a strong reduction. (D) through (J) Rtn4a and Rtn4b immunostainings at various developmental stages. At 1dpf, Rtn4a is expressed in distinct neuronal structures, including the forebrain (fb), the presumptive optic tectum (t) and the hindbrain (hb). Rtn4a is also present in the eye anlage (e), otic vesicle (ov) (D), and in muscle tissue (m) (E). At 2 dpf, Rtn4a is detected in retinal ganglion cells (RGCs) (arrow), the optic nerve (arrowhead) and the olfactory system (o) (F). At 1 dpf, Rtn4b is expressed in the same structures as Rtn4a except the muscle tissue (G). Rtn4b is also detected in the spinal cord (sc) (H). At 2 dpf, the Rtn4b signal is still present in the spinal cord (sc) and can also be seen in growing primary motor neurons (arrow) (I). In the head, Rtn4b is present in RGCs (arrow) and the optic nerve (arrowhead) (J).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
rtn4a Ab1-rtn4a WT standard conditions Prim-5 eye IHC
Ab1-rtn4a Prim-5 forebrain axon IHC
Ab1-rtn4a Prim-5 forebrain neuron IHC
Ab1-rtn4a Prim-5 hindbrain axon IHC
Ab1-rtn4a Prim-5 hindbrain neuron IHC
Ab1-rtn4a Prim-5 midbrain axon IHC
Ab1-rtn4a Prim-5 midbrain neuron IHC
Ab1-rtn4a Prim-5 muscle IHC
Ab1-rtn4a Prim-5 somite IHC
Ab1-rtn4a Long-pec cranial nerve II IHC
Ab1-rtn4a Long-pec olfactory system IHC
Ab1-rtn4a Long-pec retinal ganglion cell IHC
Ab1-rtn4a Long-pec retinal ganglion cell axon IHC
rtn4b Ab1-rtn4b WT standard conditions Prim-5 eye IHC
Ab1-rtn4b Prim-5 forebrain axon IHC
Ab1-rtn4b Prim-5 forebrain neuron IHC
Ab1-rtn4b Prim-5 hindbrain axon IHC
Ab1-rtn4b Prim-5 hindbrain neuron IHC
Ab1-rtn4b Prim-5 midbrain axon IHC
Ab1-rtn4b Prim-5 midbrain neuron IHC
Ab1-rtn4b Prim-5 spinal cord IHC
Ab1-rtn4b Long-pec cranial nerve II IHC
Ab1-rtn4b Long-pec motor neuron axon IHC
Ab1-rtn4b Long-pec olfactory system IHC
Ab1-rtn4b Long-pec retinal ganglion cell IHC
Ab1-rtn4b Long-pec retinal ganglion cell axon IHC
Ab1-rtn4b Long-pec spinal cord motor neuron IHC
Antibody Labeling Details No data available
Phenotype Details No data available
Acknowledgments:
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