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First published online 7 February 2007
doi: 10.1242/dev.02806

Development 134, 1111-1122 (2007)
Published by The Company of Biologists 2007
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Notch signaling controls the differentiation of transporting epithelia and multiciliated cells in the zebrafish pronephros

Yan Liu1, Narendra Pathak1, Albrecht Kramer-Zucker2 and Iain A. Drummond1,*

1 Nephrology Division, Massachusetts General Hospital, 149 13th Street, Charlestown, MA 02129, USA.
2 Renal Division, University Hospital Freiburg, Zentrale Klinische Forschung (ZKF), Breisacher Str. 66, 79106 Freiburg, Germany.

* Author for correspondence (e-mail: idrummon{at}receptor.mgh.harvard.edu)

Accepted 28 December 2006

Epithelial tubules consist of multiple cell types that are specialized for specific aspects of organ function. In the zebrafish pronephros, multiciliated cells (MCCs) are specialized for fluid propulsion, whereas transporting epithelial cells recover filtered-blood solutes. These cell types are distributed in a `salt-and-pepper' fashion in the pronephros, suggesting that a lateral inhibition mechanism may play a role in their differentiation. We find that the Notch ligand Jagged 2 is expressed in MCCs and that notch3 is expressed in pronephric epithelial cells. Morpholino knockdown of either jagged 2 or notch3, or mutation in mind bomb (in which Notch signaling is impaired), dramatically expands ciliogenic gene expression, whereas ion transporter expression is lost, indicating that pronephric cells are transfated to MCCs. Conversely, ectopic expression of the Notch1a intracellular domain represses MCC differentiation. Gamma-secretase inhibition using DAPT demonstrated a requirement for Notch signaling early in pronephric development, before the pattern of MCC differentiation is apparent. Strikingly, we find that jagged 2 knockdown generates extra cilia and is sufficient to rescue the kidney cilia mutant double bubble. Our results indicate that Jagged 2/Notch signaling modulates the number of multiciliated versus transporting epithelial cells in the pronephros by way of a genetic pathway involving repression of rfx2, a key transcriptional regulator of the ciliogenesis program.

Key words: Pronephros, Jagged 2, Multiciliated cell, Notch3, double bubble, mind bomb, Zebrafish


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