ZFIN ID: ZDB-FIG-111201-32
Bachmann-Gagescu et al., 2011 - The Ciliopathy Gene cc2d2a controls Zebrafish Photoreceptor Outer Segment Development Through a Role in Rab8-dependent Vesicle Trafficking. Human molecular genetics   20(20):4041-55 Full text @ Hum. Mol. Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Day 5

Fig. 3 Cc2d2a localizes to the connecting cilium/transition zone of cilia in photoreceptors and other cell types. AC′′′) Retinal cryosections from cc2d2a+/+ embryos labeled with anti-Cc2d2a (green) and anti-acetylated tubulin [red in (A–A′′′)], anti-Ift88 [red in (B–B′′′)] and anti-gamma-tubulin [red in (C–C′′′)] show Cc2d2a localization at the base of cilia (arrows). The Cc2d2a signal is apical to the basal-most acetylated tubulin signal [arrowhead in (A′′)], apical to the gamma-tubulin signal (C–C′′′) and within the Ift88-poor region that represents the connecting cilium, between the brightest Ift88 signal and a fainter, more basal signal [arrowheads in (B′ and B′′)]. Embryos are 4 d.p.f. in (A) and 5 d.p.f. in (B and C). (DG) Immunofluorescence against Cc2d2a (green) and Arl13b (red) showing Cc2d2a localization at the base of 48 h.p.f. pronephric duct cilia (D and D′), 24 h.p.f. floorplate cilia (E and E′), 72 h.p.f. olfactory pit cilia (F) and 24 h.p.f. cerebellar neuronal progenitor cilia (G). (H) High power confocal image of a neuronal progenitor cilium from a 24 h.p.f. arl13b-GFP embryo stained with antibodies against gamma-tubulin (green), Cc2d2a (red) and GFP (blue). (D–H) Single-confocal sections of whole-mount embryos. Scale bars are 6 µm in (A), 2 µm in (A′), 0.4 µm in (A′′ and A′′′), 4 µm in (B and C), 1 µm in (B′–B′′′), 0.5 µm in (C′–C′′′), 5 µm in (D and F), 10 µm in (E), 20 µm in (G) (2 µm in inset) and 0.6 µm in (H). (I) Western blot with Cc2d2a P2D9 antibody from whole wild-type, zygotic and maternal zygotic (mz) cc2d2a-/- embryo lysates at 48 h.p.f. The Cc2d2a band appears as a doublet around 190 kDa in wild-type (cc2d2a+/+) and is absent in zygotic and mz cc2d2a-/- embryos. (J) Western blot with Cc2d2a P2D9 antibody on full-length and truncated (W628*) Cc2d2a-GFP protein overexpressed in HEK293T cells. The antibody recognizes the full-length as well as the truncated Cc2d2a protein.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
arl13b Ab1-arl13b WT standard conditions Prim-5 cerebellum cilium IHC
Ab1-arl13b Prim-5 floor plate motile cilium IHC
Ab1-arl13b Long-pec pronephric duct motile cilium IHC
Ab1-arl13b Protruding-mouth ciliated olfactory receptor neuron non-motile cilium IHC
cc2d2a Ab1-cc2d2a WT standard conditions Prim-5 cerebellum ciliary transition zone IHC
Ab1-cc2d2a Prim-5 floor plate ciliary transition zone IHC
Ab1-cc2d2a Long-pec pronephric duct ciliary transition zone IHC
Ab1-cc2d2a Long-pec whole organism WB
Ab1-cc2d2a Protruding-mouth ciliated olfactory receptor neuron ciliary transition zone IHC
Ab1-cc2d2a Day 4 eye photoreceptor cell photoreceptor connecting cilium IHC
Ab1-cc2d2a Day 5 eye photoreceptor cell photoreceptor connecting cilium IHC
Ab1-cc2d2a cc2d2aw38/w38 standard conditions Long-pec Not Detected whole organism WB
Ab1-cc2d2a cc2d2aw38/w38 standard conditions Long-pec Not Detected whole organism WB
Ab1-cc2d2a hsc5Tg standard conditions Prim-5 neuron ciliary transition zone IHC
GFP hsc5Tg standard conditions Prim-5 neuron cilium IHC
ift88 WT standard conditions Day 5 eye photoreceptor cell cilium IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-tuba IHC WT standard conditions Day 4 eye photoreceptor cell axoneme
Ab1-tubg1 IHC WT standard conditions Day 5 eye photoreceptor cell ciliary basal body
IHC hsc5Tg standard conditions Prim-5 neuron ciliary basal body
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal Human molecular genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Hum. Mol. Genet.