ZFIN ID: ZDB-FIG-161212-6
Randlett et al., 2013 - Cellular requirements for building a retinal neuropil. Cell Reports   3:282-90 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Protruding-mouth

Fig. 3

BC Axons Form Presynaptic Structures without Postsynaptic Neurons

(A) WT ath5:GAP-RFP retinas stained with anti-HuC/D (to label ACs/RGCs) and anti-RibeyeA (to label ribbon synapses). Obvious punctate RibeyeA staining is visible in the IPL, whereas weak staining is seen in the BC cell bodies in the apical half of the INL and the OPL.

(B) When ACs and RGCs are absent in the ath5:GAP-RFP;ath5−/−;ptf1a−/−;ptf1aMO embryos, the weakly RibeyeA-staining BC cell bodies span the entire INL, and punctate RibeyeA staining is visible in the BC-only IPL. Scale bar in (A) and (B), 10 μm.

(C) Examples of vesicle-filled bouton-like structures (asterisks) containing ribbons near the retinal basal lamina of animals with severely reduced numbers of ACs and RGCs. Left: Higher magnification of the two such structures, presumed to be BC axonal boutons (arrowhead) shown in the inset. Right: Three consecutive sections (s1–s3) of processes in basally located neuropil in another animal. Arrows indicate ribbons juxtaposed to appositions with other processes, one of which can be another BC terminal (s1–s3).

(D and E) WT BC synapses exhibit readily apparent postsynaptic densities (thickenings, arrow) that are not seen in the mutant/morphant plexus (s1–s3).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
RFP atoh7th241/th241; ptf1asa126/sa126; cu2Tg + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth bipolar neuron IHC
Protruding-mouth photoreceptor cell IHC
Protruding-mouth retinal inner plexiform layer IHC
cu2Tg standard conditions Protruding-mouth bipolar neuron IHC
Protruding-mouth photoreceptor cell IHC
Protruding-mouth retinal inner plexiform layer IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
Ab1-ctbp2a IHC atoh7th241/th241; ptf1asa126/sa126; cu2Tg + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth bipolar neuron ribbon synapse
IHC Protruding-mouth retinal inner plexiform layer
IHC cu2Tg standard conditions Protruding-mouth bipolar neuron ribbon synapse
IHC Protruding-mouth retinal inner plexiform layer
Ab1-elavl IHC atoh7th241/th241; ptf1asa126/sa126; cu2Tg + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth Not Detected retina
IHC cu2Tg standard conditions Protruding-mouth amacrine cell
IHC Protruding-mouth retinal ganglion cell
Phenotype Details
Fish Conditions Stage Phenotype
atoh7th241/th241; ptf1asa126/sa126; cu2Tg + MO1-ptf1a + MO4-ptf1a + MO4-tp53 standard conditions Protruding-mouth amacrine cell absent, abnormal
Protruding-mouth bipolar neuron postsynaptic density absent, abnormal
Protruding-mouth retina ab1-elavl labeling absent, abnormal
Protruding-mouth retinal ganglion cell 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.