ZFIN ID: ZDB-FIG-160513-18
Zhou et al., 2015 - Gmnc Is a Master Regulator of the Multiciliated Cell Differentiation Program. Current biology : CB   25(24):3267-73 Full text @ Curr. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 16-cell to Prim-5
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage: Prim-5

Fig. 3

gmnc Is Expressed in the Kidney Tubules and Is Modulated by Notch Signaling and Foxj1a Activation

(A) End-point PCRs on cDNA extracted from 1.5, 5, 14, 18, and 24 hpf embryos, respectively, revealed that whereas the maternal contribution of the foxj1a transcript is minimal (1.5 hr), the gmnc transcript is maternally deposited. actin-b1 (actb1) was used as the loading control.

(B and C) Representative staining from in situ hybridization using a probe against gmnc showed that the gene is expressed in the developing nasal placodes (arrows) at 18 hpf (B) and is also expressed in a spotted pattern in the mid-section of the kidney ducts (asterisks) at 24 hpf (C). Scale bars, 50 ┬Ám.

(D) qPCRs were performed on uninjected or jagged2a MO-injected embryos at 24 hpf. Similar to the ciliogenic genes like foxj1a, foxj1b, and mcidas (mci), whose levels increased in the N-deficient background, gmnc transcript levels also increased moderately (by ~3-fold). Expression levels in the uninjected condition were arbitrarily assigned a value of 1. rplpo was used as an internal (loading) control. Error bars represent the SEM from two independent experiments.

(E) qPCRs were performed on non-heat-shocked or heat-shocked Tg(hsp70::foxj1a) embryos. The heat-shock treatment induced the foxj1a transgene level by ~15-fold, and the foxj1a overexpression induced the gmnc level by ~10-fold. Expression levels in the non-heat-shocked condition were arbitrarily assigned a value of 1. actin-b1 (actb1) was used as an internal (loading) control. Error bars represent the SEM from three independent experiments.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
foxj1a WT standard conditions 16-cell Not Detected whole organism RTPCR
30%-epiboly whole organism RTPCR
10-13 somites whole organism RTPCR
14-19 somites whole organism RTPCR
Prim-5 whole organism RTPCR
WT + MO5-jag2b standard conditions Prim-5 whole organism RTPCR
sq5713Tg standard conditions Prim-5 whole organism RTPCR
sq5713Tg heat shock Prim-5 whole organism RTPCR
foxj1b WT standard conditions Prim-5 whole organism RTPCR
WT + MO5-jag2b standard conditions Prim-5 whole organism RTPCR
gmnc WT standard conditions 16-cell whole organism RTPCR
30%-epiboly whole organism RTPCR
10-13 somites whole organism RTPCR
14-19 somites olfactory placode ISH
14-19 somites whole organism RTPCR
Prim-5 pronephric duct ISH
Prim-5 whole organism RTPCR
WT + MO5-jag2b standard conditions Prim-5 whole organism RTPCR
sq5713Tg standard conditions Prim-5 whole organism RTPCR
sq5713Tg heat shock Prim-5 whole organism RTPCR
mcidasl WT standard conditions Prim-5 whole organism RTPCR
WT + MO5-jag2b standard conditions Prim-5 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO5-jag2b standard conditions Prim-5 whole organism foxj1a expression increased amount, abnormal
Prim-5 whole organism foxj1b expression increased amount, abnormal
Prim-5 whole organism gmnc expression increased amount, abnormal
Prim-5 whole organism mcidasl expression increased amount, abnormal
sq5713Tg heat shock Prim-5 whole organism foxj1a expression increased amount, abnormal
Prim-5 whole organism gmnc expression increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Current biology : CB for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Curr. Biol.