ZFIN ID: ZDB-FIG-140822-21
Ishikawa et al., 2014 - TTC26/DYF13 is an intraflagellar transport protein required for transport of motility-related proteins into flagella. eLIFE   3:e01566 Full text @ Elife
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 10-13 somites to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: 5-9 somites to Long-pec

Fig. 2

ttc26 knockdown zebrafish shows phenotypes typical of defective cilia.

(A) Expression of TTC26 in control and ttc26 knockdown zebrafish. 3 dpf embryos were lysed with 1× SDS sample buffer and detected by Western blotting with TTC26 and actin antibodies. Control, AUG-MO and SP-MO show negative control, translational blocking, and splice blocking ttc26 morpholinos injected embryos, respectively. Actin antibody was used as a loading control. (B) Brightfield images of control and ttc26 knockdown fish at 3 dpf. ttc26 knockdown fish showed phenotypes typical of defective cilia. (C) Representative images of anterior region of embryos at 2 dpf. ttc26 knockdown fish showed hydrocephalus (black arrowhead), pronephric cyst (arrow), and abnormal ear otolith formation (white arrowhead). (D) Transmission electron micrographs of retina at 4 dpf. ttc26 knockdown fish lost outer segment (OS) of photoreceptor. Bar: 5 µm. (E) Transmission electron micrographs of connecting cilium in the retina. ttc26 knockdown fish still have connecting cilia (CC: arrow) despite the absence of outer segments. Bar: 500 nm. CC, connecting cilium; IS, inner segment; M, mitochondria; N, nucleus; OS, outer segment; RPE, retinal pigment epithelium.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP f1Tg control Long-pec atrium IFL
Long-pec cardiac ventricle IFL
Long-pec heart IFL
f1Tg + MO1-ttc26 control Long-pec atrium IFL
Long-pec cardiac ventricle IFL
Long-pec heart IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-actb IHC TL control 10-13 somites Kupffer's vesicle cilium
IHC 20-25 somites pronephric duct cilium
IHC TL + MO1-ttc26 control 10-13 somites Kupffer's vesicle cilium
IHC 20-25 somites pronephric duct cilium
Phenotype Details
Fish Conditions Stage Phenotype
TL + MO1-ttc26 control 5-9 somites Kupffer's vesicle cilium movement decreased frequency, abnormal
5-9 somites Kupffer's vesicle cilium movement process quality, abnormal
10-13 somites Kupffer's vesicle cilium decreased amount, abnormal
10-13 somites Kupffer's vesicle cilium decreased length, abnormal
20-25 somites pronephric duct cilium decreased length, abnormal
f1Tg + MO1-ttc26 control Long-pec heart looping process quality, abnormal
Acknowledgments:
ZFIN wishes to thank the journal eLIFE for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Elife