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ZIRC
ZFIN ID: ZDB-FIG-161212-7
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:
Antibodies:
Fish:
Condition:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Protruding-mouth to Day 4

Fig. 4

MG Are Not Required for Retinal Organization or Plexiform Layer Development

(A) ath5:GAP-RFP and vsx1:GFP show that all neural retinal cell types are present and correctly positioned in DMSO-treated control embryos and after DAPT treatment.

(B and C) Treatment with DAPT at 33 hpf completely removed MG in the retina. PKC staining is unperturbed by DAPT treatment, indicating BC differentiation is unaffected. However, DAPT-treated embryos show a complete loss of MG when stained with specific markers for GS, CRALBP, and GFAP when compared with DMSO controls.

(D) The IPL forms and sublaminates properly in the absence of MG. nyx:Gal4; UAS:MYFPQ16, and vsx1:MCeruleanQ19 fish show labeling of specific subsets of BCs that stratify in the apical (OFF) or basal (ON) domains of the IPL. Imaging of triple transgenics Q16;Q19;gfap:GFP treated with DAPT shows that the IPL still forms and BC axons sublaminate properly (arrow). Lack of gfap:GFP signal confirms that there are no MG in the areas of proper sublamination.

(E) Pre- and postsynaptic markers remain in the plexiform layers lacking MG. Staining for the presynaptic markers RibeyeA and SV2 and the postsynaptic density protein Maguk (Mag) is unperturbed in embryos lacking MG at 72 hpf. Scale bars, 10 μm (A and D) and 20 μm (B, C, and E).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
Cerulean q16bTg ; q16aTg ; q19Tg control Day 4 bipolar neuron IHC
Day 4 retinal inner plexiform layer IHC
q16bTg ; q16aTg ; q19Tg chemical treatment: DAPT Day 4 bipolar neuron IHC
Day 4 retinal inner plexiform layer IHC
GFP cu2Tg ; nns5Tg control Protruding-mouth bipolar neuron IHC
Protruding-mouth bipolar neuron axon IHC
Protruding-mouth retinal inner plexiform layer IHC
Protruding-mouth retinal outer plexiform layer IHC
cu2Tg ; nns5Tg chemical treatment: DAPT Protruding-mouth bipolar neuron IHC
Protruding-mouth bipolar neuron axon IHC
Protruding-mouth retinal inner plexiform layer IHC
Protruding-mouth retinal outer plexiform layer IHC
RFP cu2Tg ; nns5Tg control Protruding-mouth amacrine cell IHC
Protruding-mouth horizontal cell IHC
Protruding-mouth photoreceptor cell IHC
Protruding-mouth retinal ganglion cell IHC
cu2Tg ; nns5Tg chemical treatment: DAPT Protruding-mouth amacrine cell IHC
Protruding-mouth horizontal cell IHC
Protruding-mouth photoreceptor cell IHC
Protruding-mouth retinal ganglion cell IHC
YFP q16bTg ; q16aTg ; q19Tg control Day 4 bipolar neuron IHC
Day 4 retinal inner plexiform layer IHC
q16bTg ; q16aTg ; q19Tg chemical treatment: DAPT Day 4 bipolar neuron IHC
Day 4 retinal inner plexiform layer IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
Ab-SV2 IHC WT control Protruding-mouth retinal inner plexiform layer presynapse
IHC WT chemical treatment: DAPT Protruding-mouth retinal inner plexiform layer presynapse
Ab1-ctbp2a IHC WT control Protruding-mouth retinal inner plexiform layer ribbon synapse
IHC WT chemical treatment: DAPT Protruding-mouth retinal inner plexiform layer ribbon synapse
Ab1-glul IHC WT control Protruding-mouth Muller cell
IHC WT chemical treatment: DAPT Protruding-mouth Not Detected retina
Ab1-pan-MAGUK IHC WT control Protruding-mouth retinal inner plexiform layer postsynaptic density
IHC WT chemical treatment: DAPT Protruding-mouth retinal inner plexiform layer postsynaptic density
Ab2-prkcb IHC WT control Protruding-mouth bipolar neuron
IHC Protruding-mouth retinal inner plexiform layer
IHC WT chemical treatment: DAPT Protruding-mouth bipolar neuron
IHC Protruding-mouth retinal inner plexiform layer
zrf-1 IHC WT control Protruding-mouth Muller cell
IHC WT chemical treatment: DAPT Protruding-mouth Not Detected retina
IHC q16bTg ; q16aTg ; q19Tg control Day 4 Muller cell
IHC q16bTg ; q16aTg ; q19Tg chemical treatment: DAPT Day 4 Not Detected retina
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment: DAPT Protruding-mouth Muller cell absent, abnormal
Protruding-mouth retina zrf-1 labeling absent, abnormal
Protruding-mouth retina ab1-glul labeling absent, abnormal
cu2Tg ; nns5Tg chemical treatment: DAPT Protruding-mouth Muller cell absent, abnormal
q16bTg ; q16aTg ; q19Tg chemical treatment: DAPT Day 4 retina zrf-1 labeling absent, 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.