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Departments of Comparative Medicine and Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294-0019
Flk1, a receptor tyrosine
kinase for vascular endothelial growth factor (VEGF), is the earliest
known marker for endothelial precursors (angioblasts). We examined
heterozygous mice in which the Flk1
gene was partially replaced by a promoter-less
LacZ insert and used 
-galactosidase
histochemistry to view cells transcribing Flk1. In day
10 (E10) embryos, a
Flk1-positive network surrounded the metanephric blastema, and, at
E11, a vessel entered the metanephros from its ventral aspect alongside the ingrowing ureteric bud. However,
aortic branches did not engage embryonic kidneys at these time points.
In newborns, -galactosidase was localized exclusively and intensely
to endothelial cells of all vessels and glomeruli. In contrast, when
E12 kidneys grown in organ culture for
6 days were examined, only scattered Flk1-positive cells were seen,
glomeruli were unlabeled, and vessels were absent. When organ-cultured
kidneys were then grafted into wild-type anterior eye chambers,
numerous Flk1-positive endothelial cells in vessels and glomeruli were found, all stemming from the graft. Image analysis showed that grafts
with the most abundant glomerulo- and tubulogenesis were also those
with the richest expression of Flk1. We conclude that 1) kidney microvessels precede renal
artery development, 2) angioblast differentiation is arrested in organ culture but released on grafting when vasculogenesis resumes, and 3)
nephrogenesis and microvessel assembly are tightly coupled in vivo.
angiogenesis; nephrogenesis; vasculogenesis
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