ZFIN ID: ZDB-FIG-140902-103
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:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. 6

Retinal ganglion cell development in rtn4a- and rtn4b-knockdown, GFP–transgenic embryos. (A) through (I)tg(brn3c:mGFP) embryos at 3 dpf exhibit labeled retinal ganglion cells (RGCs) in the eye (arrow) and RGC axons in the optic nerve and tract as well as in the optic tectum (t). (A), (D) and (G) Lateral views of the left eye and optic tectum (anterior to the left). (B), (C), (E), (F), (H) and (I) Ventral views (anterior to the top). (A) to (C) In control morpholino (Co MO)–injected embryos, RGC axons reached the optic tectum (t) and innervated the neuropil (A). The ventral view shows the RGCs in both eyes (arrow) and the RGC axons in the optic nerve, the chiasm and the optic tract (B). (C), (F) and (I) RGCs and their axons were costained with antibodies against neurolin. RGCs from both morphants, rtn4a(D) to (F) and, to a greater extent, rtn4b(G) to (I), cover smaller areas of the retina (outlined) and their axons show aberrant pathways. (J) to (L) Quantification of eye size, the area covered by RGCs and their ratio. (M) to (O) and (P) to (R) Rescue of the rtn4a and rtn4b RGC phenotypes. Arrow points to RGCs. (S) Quantification at 3 dpf showing the reduction in the area covered by RGCs after injection of rtn4a- and rtn4b-MO1 (separately and combined) and rtn4b-MO1 and rtn4b-mRNA, as indicated under each bar. Control-MO (n = 28), rtn4a-l-MO1 (5.0 ng) (n = 16), rtn4a-l-MO1 (2.5 ng) + rtn4a-l-mRNA (100 pg) (n = 32), rtn4b-MO1 (2.5 ng) (n = 22), rtn4b-MO2 (2.5 ng) (n = 24), rtn4b-MO1 (5.0 ng) (n = 19), rtn4b-MO2 (5.0 ng) (n = 24), rtn4b-MO1 and rtn4b-MO2 (2.5 ng) (n = 27), rtn4b-MO (5.0 ng) and rtn4b-mRNA (100 pg) (n = 23) and rtn4a-l-MO1 (2.5 ng) and rtn4b-MO1 (2.5 ng) (n = 27). Scale bar = 100 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP s356tTg control Protruding-mouth cranial nerve II IFL
Protruding-mouth optic chiasm IFL
Protruding-mouth optic tectum IFL
Protruding-mouth optic tract IFL
Protruding-mouth retinal ganglion cell IFL
Protruding-mouth retinal ganglion cell axon IFL
s356tTg + MO2-rtn4b standard conditions Protruding-mouth cranial nerve II IFL
Protruding-mouth optic chiasm IFL
Protruding-mouth optic tectum IFL
Protruding-mouth optic tract IFL
Protruding-mouth retinal ganglion cell IFL
Protruding-mouth retinal ganglion cell axon IFL
s356tTg + MO9-rtn4a standard conditions Protruding-mouth cranial nerve II IFL
Protruding-mouth optic chiasm IFL
Protruding-mouth optic tectum IFL
Protruding-mouth optic tract IFL
Protruding-mouth retinal ganglion cell IFL
Protruding-mouth retinal ganglion cell axon IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab2-alcama IHC WT control Protruding-mouth cranial nerve II
IHC Protruding-mouth retinal ganglion cell
IHC Protruding-mouth retinal ganglion cell axon
IHC WT + MO2-rtn4b standard conditions Protruding-mouth cranial nerve II
IHC Protruding-mouth retinal ganglion cell
IHC Protruding-mouth retinal ganglion cell axon
IHC WT + MO9-rtn4a standard conditions Protruding-mouth cranial nerve II
IHC Protruding-mouth retinal ganglion cell
IHC Protruding-mouth retinal ganglion cell axon
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO2-rtn4b standard conditions Protruding-mouth cranial nerve II has fewer parts of type retinal ganglion cell axon, abnormal
Protruding-mouth retina has fewer parts of type retinal ganglion cell, abnormal
Protruding-mouth retinal ganglion cell axon guidance process quality, abnormal
WT + MO9-rtn4a standard conditions Protruding-mouth cranial nerve II has fewer parts of type retinal ganglion cell axon, abnormal
Protruding-mouth retina has fewer parts of type retinal ganglion cell, abnormal
Protruding-mouth retinal ganglion cell axon guidance process quality, abnormal
s356tTg + MO2-rtn4b standard conditions Protruding-mouth cranial nerve II has fewer parts of type retinal ganglion cell axon, abnormal
Protruding-mouth retina has fewer parts of type retinal ganglion cell, abnormal
Protruding-mouth retinal ganglion cell axon guidance process quality, abnormal
s356tTg + MO9-rtn4a standard conditions Protruding-mouth cranial nerve II has fewer parts of type retinal ganglion cell axon, abnormal
Protruding-mouth retina has fewer parts of type retinal ganglion cell, abnormal
Protruding-mouth retinal ganglion cell axon guidance process quality, abnormal
Acknowledgments:
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