ZFIN ID: ZDB-FIG-200722-14
Schlegel et al., 2019 - A New Zebrafish Model for CACNA2D4-Dysfunction. Investigative ophthalmology & visual science   60:5124-5135 Full text @ Invest. Ophthalmol. Vis. Sci.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Anatomical Term:
Stage: Day 5
PHENOTYPE:
Fish:
Observed In:
Stage: Day 5

Fig. 7 Ultrastructure of the photoreceptor ribbon synapse. (A) Correlative light- and electron microscopy (CLEM) on 5 dpf retinae of WT and double-KO. Cacna1fa puncta seem to be reduced in number in double-KO compared to WT, but both show correct localization of Cacna1fa to synaptic ribbons in the OPL (close up). Red: Cacna1fa antibody staining. Cyan: DAPI. Blue: pseudo-colored ribbon. White frame: close up region. Scale bar in overview corresponds to 10 μm and in close-up to 1 μm. (B) Transmission electron micrographs (TEM) of synapses in 5 dpf WT and double-KO. The presynaptic terminal is pseudocolored in yellow and the postsynaptic invaginations in red. Ribbons that are attached to the active zone are found in cones and rods of both genotypes (blue arrows), but also some that are ‘floating' or missing (red arrows). Scale bar: 0.5 μm. (C–E) Quantification of synapse and ribbon number or attachment on whole-retinal scans of three sections per genotype (one per individual). (C) The number of synapses per 100 μm in double-KO is normal, however the amount of ribbons is reduced and there are fewer ribbons attached to the active zone. (D) Ternary plot of synapse subtypes: 0 rib = empty synapses; 1 rib = synapses with one ribbon; >1 rib = synapses with more than one ribbon; grey = WT; blue = double-KO. The data from WT and double-KO form two separate clusters. The percentage of synapses harboring more than one ribbon (cones) is lower (P = 0.03) and that of empty synapses is higher (P = 0.05) in double-KO. 1-ribbon synapses do not seem to be affected. (E) The number of attached ribbons relative to the total number of ribbons is lower in double-KO retinae. Box and whisker plots: bottom and top of the box, first and third quartile; the median, line within the box; whiskers, minimum and maximum values. Significance levels: *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cacna1fa Ab1-cacna1fa TU standard conditions Day 5 eye photoreceptor cell photoreceptor ribbon synapse IHC
Ab1-cacna1fa cacna2d4azh5/zh5; cacna2d4bzh6/zh6(TU) standard conditions Day 5 eye photoreceptor cell photoreceptor ribbon synapse IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
cacna2d4azh5/zh5; cacna2d4bzh6/zh6(TU) standard conditions Day 5 eye photoreceptor cell synaptic ribbon separated from eye photoreceptor cell photoreceptor ribbon synapse, abnormal
Day 5 retinal cone cell development decreased process quality, abnormal
Day 5 retinal outer plexiform layer has fewer parts of type eye photoreceptor cell synaptic ribbon, abnormal
Day 5 retinal outer plexiform layer has normal numbers of parts of type eye photoreceptor cell photoreceptor ribbon synapse, normal
Acknowledgments:
ZFIN wishes to thank the journal Investigative ophthalmology & visual science for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Invest. Ophthalmol. Vis. Sci.