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ZFIN ID: ZDB-FIG-150910-29
Toruń et al., 2015 - Endocytic Adaptor Protein Tollip Inhibits Canonical Wnt Signaling. PLoS One   10:e0130818 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Shield to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 8 zTollip contributes to the canonical Wnt pathway in embryonic development.

(A-E) Embryos overexpressing ztollip (B) or injected with either ztollip morpholinos (MOs) (C), or β-catenin2 MO (D), or both (E) were analyzed for their morphological phenotype at 48 hpf. Typical phenotypes are shown. Lateral views with head to the left. Injection of ztollip mRNA led to reduction of posterior structures (arrow in B) compared with wild-type, control embryos (A). Knockdown of ztollip resulted in tail curvature (arrow in C) and pericardial oedema (asterisk in representative embryo in inset of C). Injection of β-catenin2 MO caused morphological defects similar to those found in embryos overexpressing ztollip (compare B and D). Tail curvature and pericardial oedema in embryos overexpressing ztollip could be rescued by injection of 2 ng of β-catenin2 MO (compare B and E). (F-R) In situ hybridization using different mRNA probes. Embryos injected with ztollip mRNA showed expanded expression of both the dorsal mesendodermal marker goosecoid (gsc) at 6 hpf (shield stage) (brackets in G) and the dorsal marker notail (ntl) at tail bud stage (brackets in I) compared to uninjected embryos (F and H, respectively). Expression of the cardiac marker cmlc2 (J, K) indicated the failure of the heart to form the loop in ztollip morphants (asterisk in J) compared to wild-type embryos (asterisk in K) at 48 hpf. Expression of fgf8 in the tail was reduced or absent in embryos overexpressing ztollip (arrow in M), while embryos injected with ztollip MOs showed an increase in fgf8 expression in the brain and in the tail (arrows in O) compared to uninjected, control embryos (L) at 24 hpf. Embryos injected with 8 ng β-catenin2 MO exhibited reduced fgf8 expression (N). Blocking endocytosis by injecting embryos with 4 ng rab5a MO or by expressing the Dnm2-K44A displayed no clear effect on fgf8 expression (arrows in P and Q). ztollip overexpression together with the Dnm2-K44A mutant showed a Tollip-like phenotype on the expression of fgf8 (arrow in R).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
fgf8a AB control Prim-5 brain ISH
Prim-5 caudal fin ISH
AB + MO1-ctnnb2 control Prim-5 brain ISH
Prim-5 caudal fin ISH
AB + MO1-rab5aa control Prim-5 brain ISH
Prim-5 caudal fin ISH
AB + MO1-tollip control Prim-5 brain ISH
Prim-5 caudal fin ISH
AB + MO2-tollip control Prim-5 brain ISH
Prim-5 caudal fin ISH
AB + MO3-tollip control Prim-5 brain ISH
Prim-5 caudal fin ISH
gsc AB control Shield hypoblast dorsal region ISH
myl7 AB control Long-pec heart ISH
AB + MO1-tollip control Long-pec heart ISH
AB + MO2-tollip control Long-pec heart ISH
AB + MO3-tollip control Long-pec heart ISH
tbxta AB control Bud whole organism dorsal region ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO1-ctnnb2 control Prim-5 brain fgf8a expression decreased amount, abnormal
Prim-5 caudal fin fgf8a expression decreased amount, abnormal
AB + MO1-ctnnb2 standard conditions Long-pec whole organism posterior region malformed, abnormal
AB + MO1-tollip control Prim-5 brain fgf8a expression increased amount, abnormal
Prim-5 caudal fin fgf8a expression increased amount, abnormal
AB + MO1-tollip standard conditions Long-pec caudal fin curved, abnormal
Long-pec heart looping decreased occurrence, abnormal
Long-pec pericardium edematous, abnormal
AB + MO2-tollip control Prim-5 brain fgf8a expression increased amount, abnormal
Prim-5 caudal fin fgf8a expression increased amount, abnormal
AB + MO2-tollip standard conditions Long-pec caudal fin curved, abnormal
Long-pec heart looping decreased occurrence, abnormal
Long-pec pericardium edematous, abnormal
AB + MO3-tollip control Prim-5 brain fgf8a expression increased amount, abnormal
Prim-5 caudal fin fgf8a expression increased amount, abnormal
AB + MO3-tollip standard conditions Long-pec caudal fin curved, abnormal
Long-pec heart looping decreased occurrence, abnormal
Long-pec pericardium edematous, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One