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Fig. 2. Nephrogenesis arrests and cells in the peripheral zone die in the absence
of FGF8. Marker analysis in (A,B,Q-T) cryosections or (C-P,U,V) vibratome
sections of kidneys at the stages indicated. (A-L,U,V) In situ hybridization
for the genes indicated. The Fgf8 (FL) probe we used contained the
full-length coding sequence, and therefore detected Fgf8 RNA produced
by the Fgf8null allele. (M-P) Immunofluorescence assays
for PAX2 (green) to identify the developing nephrons and collecting ducts,
co-stained with LysoTracker (LysoT, red) to identify regions containing dying
cells. (Q-T) Immunofluorescence assays for PAX2 (green) and Calbindin (CB,
blue), which identifies collecting ducts, and for TUNEL staining (red), which
detects dying cells. Arrowheads point to nascent nephrons, which are present
in normal kidneys and also in Fgf8-MM-KO kidneys at E13.5 (A,B) and
E14.5 (Q,R), but not at E16.5 (S,T). Note that the nascent nephrons in
Fgf8-MM-KO kidneys (B,R) have formed an epithelial structure
surrounding a lumen, i.e. they have reached the renal vesicle stage.
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