Exogenous Slit2 does not affect ureteric branching or nephron formation during kidney development.
In an attempt to elucidate the role of Slit2 in vertebrate kidney development, the effect of adding exogenous human Slit2 protein (hSlit2) to developing murine metanephric kidney explants was examined. To confirm the activity of the recombinant Slit2 protein, neurons from 8 day old chick sympathetic nerve chain dorsal root ganglia were cultured with hSlit2 protein, which induced significant neurite branching and outgrowth. Using kidney explants as a model system, metanephric development in the presence of hSlit2 protein was examined. Addition of hSlit2 up to a final concentration of 1 microg/ml had no detectable effect on the formation of nephrons or on branching morphogenesis of the ureteric tree after 2 or 4 days in culture, as assessed via immunofluorescence for the markers WT1 and calbindin 28K respectively. Similarly, maturation of the nephrogenic mesenchyme occurred in a phenotypically normal fashion. In situ analysis of the Slit receptors, Robo1 and Robo2, the vasculogenic markers VEGFA and Flk-1, and the stromal cell marker BF2 displayed no difference in comparison to controls.
Piper
Michael
M
Institute for Molecular Bioscience, and Department of Biochemistry, University of Queensland, Brisbane, Australia.
Nurcombe
Victor
V
Wilkinson
Lorine
L
Little
Melissa
M
D000818:Animals; D019556:COS Cells; D064026:Calbindins; D002642:Chick Embryo; D016228:Endothelial Growth Factors / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D008222:Lymphokines / Q000378:metabolism; D008648:Mesoderm / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009399:Nephrons / Q000196:embryology*; D009419:Nerve Tissue Proteins / Q000378:metabolism* / Q000502:physiology*; D009474:Neurons / Q000378:metabolism; D010641:Phenotype; D017434:Protein Structure, Tertiary; D011971:Receptors, Immunologic / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D014513:Ureter / Q000196:embryology*; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D042442:Vascular Endothelial Growth Factors; D000097905:Roundabout Proteins; D000098552:Slit Homolog 2 Protein
D064026:Calbindins; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D009419:Nerve Tissue Proteins; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; D064030:S100 Calcium Binding Protein G; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2; D042442:Vascular Endothelial Growth Factors; D000097905:Roundabout Proteins; D000098552:Slit Homolog 2 Protein