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First published online September 2, 2003
doi: 10.1242/10.1242/dev.00697
RESEARCH ARTICLE: DEVELOPMENT AND DISEASE |
-Secretase activity is dispensable for mesenchyme-to-epithelium transition but required for podocyte and proximal tubule formation in developing mouse kidney
1 Department of Molecular Biology and Pharmacology, Washington University School
of Medicine, Box 8103, 660 South Euclid Avenue, St Louis, MO 63110, USA
2 Department of Medicine, Renal Division, Washington University School of
Medicine, Box 8126, 660 South Euclid Avenue, St Louis, MO 63110, USA
3 Department of Physiology, The University of Texas Southwestern Medical Center
at Dallas, 5323 Harry Hines Blvd, Dallas, TX 75390, USA
4 Department of Pathology, The University of Alabama at Birmingham, 1530 3rd
Avenue, Birmingham, AL 35294-0019, USA
5 Department of Medicine, Division of Dermatology, Washington University School
of Medicine, Box 8103, 660 South Euclid Avenue, St Louis, MO 63110, USA
* Authors for correspondence (e-mail: kopan{at}molecool.wustl.edu and minerj{at}pcg.wustl.edu)
Accepted 1 July 2003
Notch signaling is involved in pronephros development in Xenopus
and in glomerulogenesis in mice. However, owing to early lethality in mice
deficient for some Notch pathway genes and functional redundancy for others, a
role for Notch signaling during early stages of metanephric development has
not been defined. Using an antibody specific to the N-terminal end of
-secretase-cleaved Notch1, we found evidence for Notch1 activation in
the comma and S-shaped bodies of the mouse metanephros. We therefore cultured
mouse metanephroi in the presence of a -secretase inhibitor,
N-S-phenyl-glycine-t-butyl ester (DAPT), to block
Notch signaling. We observed slightly reduced ureteric bud branching but
normal mesenchymal condensation and expression of markers indicating that
mesenchyme induction had occurred. However, fewer renal epithelial structures
were observed, with a severe deficiency in proximal tubules and glomerular
podocytes, which are derived from cells in which activated Notch1 is normally
present. Distal tubules were present but in reduced numbers, and this was
accompanied by an increase in intervening, non-epithelial cells. After a
transient 3-day exposure to DAPT, proximal tubules expanded, but podocyte
differentiation failed to recover after removal of DAPT. These observations
suggest that -secretase activity, probably through activation of Notch,
is required for maintaining a competent progenitor pool as well as for
determining the proximal tubule and podocyte fates.
Key words: Metanephric culture, Notch, gamma-secretase, DAPT, Wt1, Mouse
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