ZFIN ID: ZDB-FIG-161212-4
Randlett et al., 2013 - Cellular requirements for building a retinal neuropil. Cell Reports   3:282-90 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Protruding-mouth to Day 5

Fig. 2

Simultaneous Removal of BC Partner Neurons (RGCs and ACs) Does Not Prevent BC Axons from Forming a Sublaminated Neuropil

(A and B) Despite the loss of ACs, a phalloidin-rich IPL forms in the ptf1a−/−;ptf1aMO retina.

(C) WT retina with the actin-rich IPL shown by phalloidin (arrow) and BC terminals labeled with vsx1:GFP.

(D) Phalloidin staining demonstrates that in the AC/RGC-free retina (ath5−/−;ptf1a−/− ;ptf1aMO), the IPL is actin rich and positioned along the basal surface of the retina (arrow).

(E) The thickness of the IPL was measured for each genotype at 5 dpf, demonstrating it is significantly thinner after AC removal (ptf1a−/−;ptf1aMOs), and thicker after RGC removal (ath5−/−). One-way ANOVA and Tukey post hoc tests; ∗∗p < 0.01; error bars ± 1 SEM. The number of retinas measured is given within the bars.

(F) In the WT retina, Q16-labeled BCs stratify basally to Q19-labeled BCs in the IPL.

(G) After the removal of many ACs and RGCs by ptf1aMO and ath5MO injections (G), the typical Q16-basal, Q19-apical pattern is apparent in many areas.

(H) Inset in (F).

(I) Inset in (G).

(J) Line intensity profile measurements from 45 regions (three different retinas/animals) demonstrates the basal enrichment of Q16 signal and apical enrichment of Q19 signal in ath5;ptf1a morphants. Scale bars, 20 μm.

See also Figures S2 and S3.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
Cerulean q16bTg ; q16aTg ; q19Tg standard conditions Day 4 bipolar neuron IHC
Day 4 bipolar neuron axon IHC
Day 4 retinal inner plexiform layer IHC
q16bTg ; q16aTg ; q19Tg + MO1-atoh7 + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Day 4 bipolar neuron IHC
Day 4 bipolar neuron axon IHC
Day 4 retinal inner plexiform layer IHC
GFP atoh7th241/th241; ptf1asa126/sa126; nns5Tg + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth bipolar neuron IHC
nns5Tg standard conditions Protruding-mouth bipolar neuron IHC
Protruding-mouth bipolar neuron axon IHC
Protruding-mouth retinal inner plexiform layer IHC
YFP q16bTg ; q16aTg ; q19Tg standard conditions Day 4 bipolar neuron IHC
Day 4 bipolar neuron axon IHC
Day 4 retinal inner plexiform layer IHC
q16bTg ; q16aTg ; q19Tg + MO1-atoh7 + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Day 4 bipolar neuron IHC
Day 4 bipolar neuron axon IHC
Day 4 retinal inner plexiform layer IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-elavl IHC WT standard conditions Protruding-mouth amacrine cell
IHC Protruding-mouth retinal ganglion cell
IHC ptf1asa126/sa126 + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth retinal ganglion cell
Phenotype Details
Fish Conditions Stage Phenotype
atoh7th241/th241; ptf1asa126/sa126; nns5Tg + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth amacrine cell absent, abnormal
Protruding-mouth retinal ganglion cell absent, abnormal
Protruding-mouth retinal inner plexiform layer position, abnormal
Day 5 retinal inner plexiform layer decreased thickness, abnormal
nns5Tg ; atoh7th241/th241 standard conditions Day 5 retinal inner plexiform layer increased thickness, abnormal
ptf1asa126/sa126 + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth amacrine cell absent, abnormal
Protruding-mouth retinal inner plexiform layer decreased thickness, abnormal
ptf1asa126/sa126; nns5Tg + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Day 5 retinal inner plexiform layer decreased thickness, abnormal
q16bTg ; q16aTg ; q19Tg + MO1-atoh7 + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Day 4 retinal inner plexiform layer Cerulean expression spatial pattern, abnormal
Day 4 retinal inner plexiform layer YFP expression spatial pattern, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.