ZFIN ID: ZDB-FIG-150326-19
Li et al., 2014 - Zebrafish nephrogenesis is regulated by interactions between retinoic acid, mecom, and Notch signaling. Developmental Biology   386(1):111-122 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagents:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagents:
Observed In:
Stage: Prim-5

Fig. 7

mecom acts upstream of Notch signaling to modulate MCC differentiation and regulate the MCC domain. (A) Wild type embryos treated with 100 µM DAPT showed a significant increase of MCC number without ectopic MCC formation. mecom knockdown led to a caudal expansion of MCC in the DL and PD, and exhibited increased MCC density compared to wild types. A similar condensed MCC arrangement could also be seen in mecom morphants treated with DAPT. The overexpression of Notch1a resulted in decreased MCCs in heatshock induced Tg(hsp70:gal4; uas:notch1a-intra) embryos. Ectopic MCC formation associated with mecom knockdown was abolished by Notch signaling activation, though the domain of MCCs was still expanded. (B) Differentiated MCCs at 24 hpf under 10× magnification in a single nephron from wild types, wild types treated with DAPT, mecom morphants and morphants treated with DAPT, and finally wild types and mecom morphants with NICD overexpression. Note: MCCs displayed a condensed organization/cluster pattern in DAPT-treated wild type and mecom morphant pronephros, while DAPT treatment in mecom morphants failed to induce further MCC density. Arrows indicate large MCC aggregates observed in DAPT-treated wild types, mecom morphants, and DAPT-treated mecom morphants, which were absent from the wild type pronephros. For each experiment, at least 20 embryos were examined. (C) Quantification of MCC density in wild types, wild type embryos treated with DAPT, mecom morphants, and mecom morphants treated with DAPT. The Student t-test revealed significant increase of MCC density in DAPT treated wild types and mecom morphants relative to untreated wild types (***p=0.0005). Alterations of MCC density between mecom morphants and morphants treated with DAPT did not reach statistical significance. For each experiment, at least 20 embryos were examined.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
odf3b WT control Prim-5 pronephros multi-ciliated epithelial cell ISH
WT chemical treatment: pharmaceutical Prim-5 pronephros multi-ciliated epithelial cell ISH
WT + MO3-mecom + MO4-mecom chemical treatment: DAPT Prim-5 pronephros multi-ciliated epithelial cell ISH
WT + MO3-mecom + MO4-mecom standard conditions Prim-5 pronephros multi-ciliated epithelial cell ISH
kca3Tg; kca4Tg heat shock Prim-5 pronephros multi-ciliated epithelial cell ISH
kca3Tg; kca4Tg + MO3-mecom + MO4-mecom heat shock Prim-5 pronephros multi-ciliated epithelial cell ISH
smyhc1 WT control Prim-5 somite ISH
WT chemical treatment: pharmaceutical Prim-5 somite ISH
WT + MO3-mecom + MO4-mecom chemical treatment: DAPT Prim-5 somite ISH
WT + MO3-mecom + MO4-mecom standard conditions Prim-5 somite ISH
kca3Tg; kca4Tg heat shock Prim-5 somite ISH
kca3Tg; kca4Tg + MO3-mecom + MO4-mecom heat shock Prim-5 somite ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment: pharmaceutical Prim-5 pronephros multi-ciliated epithelial cell aggregated, abnormal
Prim-5 pronephros multi-ciliated epithelial cell increased amount, abnormal
WT + MO3-mecom + MO4-mecom chemical treatment: DAPT Prim-5 pronephros multi-ciliated epithelial cell aggregated, abnormal
Prim-5 pronephros multi-ciliated epithelial cell increased amount, abnormal
WT + MO3-mecom + MO4-mecom standard conditions Prim-5 pronephros multi-ciliated epithelial cell aggregated, abnormal
Prim-5 pronephros multi-ciliated epithelial cell increased amount, abnormal
kca3Tg; kca4Tg heat shock Prim-5 pronephros multi-ciliated epithelial cell decreased amount, abnormal
kca3Tg; kca4Tg + MO3-mecom + MO4-mecom heat shock Prim-5 pronephros multi-ciliated epithelial cell amount, normal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Biology, 386(1), Li, Y., Cheng, C.N., Verdun, V.A., and Wingert, R.A., Zebrafish nephrogenesis is regulated by interactions between retinoic acid, mecom, and Notch signaling, 111-122, Copyright (2014) with permission from Elsevier. Full text @ Dev. Biol.