ZFIN ID: ZDB-FIG-111201-57
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 Days 7-13
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Days 7-13

Fig. 4 cc2d2a is not required for ciliogenesis. (A and B) Connecting cilia (brackets) in wild-type (A) and cc2d2a-/- (B) photoreceptors demonstrated by transmission electron microscopy. (C and D) Apical view of ciliary axonemes labeled with acetylated tubulin antibodies (red) in wild-type (C) and cc2d2a-/- (D) whole-mount eyes in which cone outer segments are labeled with tg(TaCP:MCFP). (E and F) Peripheral views of the same eyes as in (C and D) showing only the acetylated tubulin staining. Note the shorter axonemes in cc2d2a-/- eyes (F) compared with wild-type (E). (G and H) Ciliary axonemes labeled with acetylated tubulin (red) and Ift88 (green) antibodies in wild-type (G) and cc2d2a-/- (H) retinal cryosections. Brackets highlight axonemal length. Embryos are 5 d.p.f. in (A–F) and 7 d.p.f. in (G and H). (IN) Immunofluorescence with Arl13b antibody (red) highlighting cilia and with gamma-tubulin antibody (green) highlighting basal bodies in 24 h.p.f. whole-mount mz cc2d2a+/- (I, K and M) and mz cc2d2a-/- (J, L and N) embryos showing retinal neuronal progenitor cilia (arrowheads in insets of I and J), cerebellar neuronal progenitors (arrowheads in insets of K and L) and floorplate cilia (M and N). (O and P) Pronephric duct cilia stained with acetylated tubulin (red) and gamma-tubulin (green) antibodies in 48 h.p.f. whole-mount mz cc2d2a+/- (O) and mz cc2d2a-/- (P) embryos. The anterior–posterior axis is indicated by the arrows. (Q and R) Acetylated tubulin antibody staining of olfactory pit cilia at 3 d.p.f. in whole-mount wild-type (Q) and cc2d2a-/- (R) embryos. (I–L) and (Q and R) Single-confocal sections. (C–F) and (M–P) Projected stacks of confocal sections. Scale bars are 500 nm in (A and B), 5 µm in (C and D), 10 µm in (E and F), 4 µm in (G and H), 2 µm in (I–L), 10 µm in (M and N) and 5 µm in (O–R).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
arl13b Ab1-arl13b cc2d2aw38/+ standard conditions Prim-5 cerebellum cilium IHC
Ab1-arl13b Prim-5 eye photoreceptor cell cilium IHC
Ab1-arl13b Prim-5 floor plate motile cilium IHC
Ab1-arl13b Long-pec pronephric duct motile cilium IHC
Ab1-arl13b cc2d2aw38/w38 standard conditions Prim-5 cerebellum cilium IHC
Ab1-arl13b Prim-5 eye photoreceptor cell cilium IHC
Ab1-arl13b Prim-5 floor plate motile cilium IHC
Ab1-arl13b Long-pec pronephric duct motile cilium IHC
CFP cc2d2aw38/+; w62Tg standard conditions Day 5 retinal cone cell photoreceptor outer segment membrane IFL
cc2d2aw38/w38; w62Tg standard conditions Day 5 retinal cone cell photoreceptor outer segment membrane IFL
w62Tg standard conditions Day 5 retinal cone cell photoreceptor outer segment membrane IFL
ift88 WT standard conditions Days 7-13 eye photoreceptor cell cilium IHC
cc2d2aw38/+ standard conditions Days 7-13 eye photoreceptor cell cilium IHC
cc2d2aw38/w38 standard conditions Days 7-13 eye photoreceptor cell cilium IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-tuba IHC WT standard conditions Protruding-mouth olfactory pit non-motile cilium
IHC Days 7-13 eye photoreceptor cell axoneme
IHC cc2d2aw38/+ standard conditions Long-pec pronephric duct motile cilium
IHC cc2d2aw38/+ standard conditions Protruding-mouth olfactory pit non-motile cilium
IHC Days 7-13 eye photoreceptor cell axoneme
IHC cc2d2aw38/+; w62Tg standard conditions Day 5 eye photoreceptor cell axoneme
IHC cc2d2aw38/w38 standard conditions Long-pec pronephric duct motile cilium
IHC cc2d2aw38/w38 standard conditions Protruding-mouth olfactory pit non-motile cilium
IHC Days 7-13 eye photoreceptor cell axoneme
IHC cc2d2aw38/w38; w62Tg standard conditions Day 5 eye photoreceptor cell axoneme
IHC w62Tg standard conditions Day 5 eye photoreceptor cell axoneme
Ab1-tubg1 IHC cc2d2aw38/+ standard conditions Prim-5 cerebellum ciliary basal body
IHC Prim-5 eye photoreceptor cell ciliary basal body
IHC Prim-5 floor plate ciliary basal body
IHC Long-pec pronephric duct ciliary basal body
IHC cc2d2aw38/w38 standard conditions Prim-5 cerebellum ciliary basal body
IHC Prim-5 eye photoreceptor cell ciliary basal body
IHC Prim-5 floor plate ciliary basal body
IHC Long-pec pronephric duct ciliary basal body
Phenotype Details
Fish Conditions Stage Phenotype
cc2d2aw38/+ standard conditions Prim-5 cerebellum cilium length, normal
Prim-5 cilium assembly process quality, normal
Prim-5 floor plate motile cilium length, normal
Long-pec cilium assembly process quality, normal
Long-pec pronephric duct motile cilium length, normal
cc2d2aw38/+ standard conditions Protruding-mouth cilium assembly process quality, normal
Protruding-mouth olfactory pit non-motile cilium length, normal
Days 7-13 cilium assembly process quality, normal
Days 7-13 eye photoreceptor cell axoneme length, normal
cc2d2aw38/+; w62Tg standard conditions Day 5 cilium assembly process quality, normal
Day 5 eye photoreceptor cell axoneme length, normal
cc2d2aw38/w38 standard conditions Prim-5 cerebellum cilium length, normal
Prim-5 cilium assembly process quality, normal
Prim-5 floor plate motile cilium length, normal
Long-pec cilium assembly process quality, normal
Long-pec pronephric duct motile cilium length, normal
cc2d2aw38/w38 standard conditions Protruding-mouth cilium assembly process quality, normal
Protruding-mouth olfactory pit non-motile cilium length, normal
Days 7-13 eye photoreceptor cell axoneme decreased length, abnormal
cc2d2aw38/w38; w62Tg standard conditions Day 5 eye photoreceptor cell axoneme decreased length, abnormal
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.