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Fig. 10. Enhanced ciliogenesis and double bubble-mutant rescue by
inhibition of Jagged/Notch signaling. (A) Wild-type double
bubble (dbb) heterozygotes viewed in confocal projections show
alternating thick acetylated tubulin-positive bundles of cilia (arrowheads) in
pronephric lumens (individual tubules are marked with asterisks in A-D).
(B) jagged 2 exon 20-morpholino injection results in enhanced
ciliogenesis, seen here in a single pronephric tubule with a slightly expanded
lumen. (C) A single dbb-mutant pronephric lumen showing loss
or shortening of cilia bundles (arrowheads), and a grossly dilated lumen
(*). (D) dbb-mutant homozygotes injected with
jagged 2 exon 20 morpholino recover normal tubule morphology (paired
tubules marked with asterisks) and show a dramatic recovery of ciliogenesis.
(E) dbb-mutant homozygote at 2.5 dpf showing bilateral cyst
formation in the proximal pronephros (arrow). (F) dbb-mutant
homozygote injected with jagged 2 exon 20 morpholino at the one-cell
stage, showing the absence of cyst formation (arrow; mutant rescue).
(G) Histological section of dbb-homozygote pronephros showing
a cystic pronephric tubule (*), dilated pronephric tubules (arrow)
and edema (#). (H) Histological section of a dbb homozygote
injected with jagged 2 exon 20 morpholino at the one-cell stage,
showing complete absence of cystic pathology and edema (arrow,
wild-type-appearing pronephric tubules; *, normal glomerular
structure). (I) Control DMSO-treated dbb-mutant embryo showing
cystic proximal pronephros (*). (J) DAPT treatment (100
µM started at 9.5 hpf) of dbb homozygotes eliminates cystic
distension of the pronephros (*, glomerulus; arrow, pronephric
tubule). In these experiments, dbb homozygotes were identified
independently of cyst formation by the characteristic mutant ventral axis
curvature in 25% of embryos from heterozygote pair matings.
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