ZFIN ID: ZDB-FIG-151214-29
Dutta et al., 2015 - nlz1 Is required for cilia formation in zebrafish embryogenesis. Developmental Biology   406(2):203-11 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Condition:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 90%-epiboly to 20-25 somites
PHENOTYPE:
Fish:
Condition:
Knockdown Reagents:
Observed In:
Stage Range: 90%-epiboly to Prim-5

Fig. 4

Nlz1 is downstream of Foxj1a and is regulated by canonical Wnt signaling. (A)–(D), (A′)–(D′) nlz1 expression in foxj1a morphants (B, B′, n=48/50), wnt8a morphants (C, C′, n=52/55), and LiCl treated foxj1morphants embryos (D, D′, n=25/28) compared to control MO (A, A′, n=30/30), as seen in whole mount in situ hybridization of embryos at 1S stage. (E)–(F) Cilia are labeled with anti-acetylated α tubulin antibody (green) in the KV at the 5S stage. Complete lack of KV cilia was observed in foxj1a morphants (E, n=20/20); injection of nlz1 mRNA in foxj1a morphants rescues KV cilia (F, n=12/30). Quantification of expression patterns of cmlc2 (G), and lft2 (H) in Foxj1a MO (cmlc2, n=50; lft2, n=50) and Foxj1a MO+nlz1mRNA (cmlc2, n=50; lft2, n=50) injected embryos. (A)–(F) dorsal view, (A)–(D) anterior dorsal, (A′)–(D′) posterior dorsal, (E), and (F) posterior to the top. (H), (J) Embryos were treated with LiCl as specified. (I)–(L) Whole mount in situ hybridization at 95% epiboly (I), (J) and 21 h (K), (L) resulted in expansion of nlz1 expression (J, n=45/50; L, n=15/19) compared to control (I n=40,K n=40, without LiCl). (I), (J) dorsal view, (K), (L) lateral view, anterior to left. (M) Nlz1 repressed Wnt/β-catenin signaling activated by either LiCl or β-catenin in a TOPflash luciferase assay. RLU, relative luciferase units. Ab: absent; Bi: bilateral, M: midline; R: right; L: left. Scale bar (black) in (A)–(D′), 100 µm; (I)–(J), 60 µm; (K)–(L), 75 µm. Scale bar (white) in (E)–(F), 50 µm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
znf703 WT control 1-4 somites anterior neural keel ISH
1-4 somites Kupffer's vesicle ISH
1-4 somites whole organism posterior region ISH
WT standard conditions 90%-epiboly anatomical structure ISH
20-25 somites anatomical structure ISH
WT chemical treatment: lithium chloride 90%-epiboly anatomical structure ISH
20-25 somites anatomical structure ISH
WT + MO1-wnt8a standard conditions 1-4 somites Not Detected whole organism ISH
WT + MO2-foxj1a standard conditions 1-4 somites anterior neural keel ISH
1-4 somites Kupffer's vesicle ISH
1-4 somites whole organism posterior region ISH
WT + MO2-foxj1a chemical treatment: lithium chloride 1-4 somites anterior neural keel ISH
1-4 somites Kupffer's vesicle ISH
1-4 somites whole organism posterior region ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment: lithium chloride 90%-epiboly anatomical structure znf703 expression increased distribution, abnormal
20-25 somites anatomical structure znf703 expression increased distribution, abnormal
WT + MO1-wnt8a standard conditions 1-4 somites whole organism znf703 expression absent, abnormal
WT + MO2-foxj1a standard conditions 1-4 somites Kupffer's vesicle znf703 expression decreased distribution, abnormal
1-4 somites whole organism posterior region znf703 expression decreased distribution, abnormal
5-9 somites Kupffer's vesicle motile cilium absent, abnormal
Prim-5 determination of heart left/right asymmetry disrupted, abnormal
Prim-5 determination of left/right symmetry disrupted, abnormal
WT + MO2-foxj1a chemical treatment: lithium chloride 1-4 somites Kupffer's vesicle znf703 expression amount, ameliorated
1-4 somites whole organism posterior region znf703 expression amount, ameliorated
Acknowledgments:
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Reprinted from Developmental Biology, 406(2), Dutta, S., Sriskanda, S., Boobalan, E., Alur, R.P., Elkahloun, A., Brooks, B.P., nlz1 Is required for cilia formation in zebrafish embryogenesis, 203-11, Copyright (2015) with permission from Elsevier. Full text @ Dev. Biol.