FIGURE

Fig. 4

ID

ZDB-FIG-161212-7

Publication

Randlett et al., 2013 - Cellular requirements for building a retinal neuropil

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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:MYFP^Q16, and vsx1:MCerulean^Q19 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).

Expression Data

Genes:	Cerulean GFP RFP YFP
Antibodies:	Ab-SV2 Ab1-ctbp2a Ab1-glul Ab1-pan-MAGUK Ab2-prkcb zrf-1
Fish:	WT cu2Tg; nns5Tg q16aTg; q16bTg; q19Tg
Condition:	chemical treatment: DAPT
Anatomical Terms:	amacrine cell bipolar neuron bipolar neuron axon horizontal cell Muller cell photoreceptor cell retinal ganglion cell retinal inner plexiform layer retinal inner plexiform layer postsynaptic density retinal inner plexiform layer presynapse retinal inner plexiform layer ribbon synapse retinal outer plexiform layer
Stage Range:	Protruding-mouth to Day 4

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT cu2Tg; nns5Tg q16aTg; q16bTg; q19Tg
Condition:	chemical treatment: DAPT
Observed In:	Muller cell retina
Stage Range:	Protruding-mouth to Day 4

Phenotype Detail

Acknowledgments

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