title: Renal blood flow and oxygenation drive nephron progenitor differentiation.
abstract: During kidney development, the vasculature develops via both angiogenesis (branching from major vessels) and vasculogenesis (de novo vessel formation). The formation and perfusion of renal blood vessels are vastly understudied. In the present study, we investigated the regulatory role of renal blood flow and O2 concentration on nephron progenitor differentiation during ontogeny. To elucidate the presence of blood flow, ultrasound-guided intracardiac microinjection was performed, and FITC-tagged tomato lectin was perfused through the embryo. Kidneys were costained for the vasculature, ureteric epithelium, nephron progenitors, and nephron structures. We also analyzed nephron differentiation in normoxia compared with hypoxia. At embryonic day 13.5 (E13.5), the major vascular branches were perfused; however, smaller-caliber peripheral vessels remained unperfused. By E15.5, peripheral vessels started to be perfused as well as glomeruli. While the interior kidney vessels were perfused, the peripheral vessels (nephrogenic zone) remained unperfused. Directly adjacent and internal to the nephrogenic zone, we found differentiated nephron structures surrounded and infiltrated by perfused vessels. Furthermore, we determined that at low O2 concentration, little nephron progenitor differentiation was observed; at higher O2 concentrations, more differentiation of the nephron progenitors was induced. The formation of the developing renal vessels occurs before the onset of blood flow. Furthermore, renal blood flow and oxygenation are critical for nephron progenitor differentiation.
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Nephrology, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania;
authors.forename: Christopher
authors.initials: C
authors.lastname: Rymer
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Pediatric General and Thoracic Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, and Rangos Research Center, Pittsburgh, Pennsylvania; and.
authors.forename: Jose
authors.initials: J
authors.lastname: Paredes
authors.affiliation: The Centre of Excellence in Cell-Extracellular Matrix Research, Oulu, Finland.
authors.forename: Kimmo
authors.initials: K
authors.lastname: Halt
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Nephrology, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania;
authors.forename: Caitlin
authors.initials: C
authors.lastname: Schaefer
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Pediatric General and Thoracic Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, and Rangos Research Center, Pittsburgh, Pennsylvania; and.
authors.forename: John
authors.initials: J
authors.lastname: Wiersch
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Pediatric General and Thoracic Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, and Rangos Research Center, Pittsburgh, Pennsylvania; and.
authors.forename: Guangfeng
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Pediatric General and Thoracic Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, and Rangos Research Center, Pittsburgh, Pennsylvania; and.
authors.forename: Douglas
authors.initials: D
authors.lastname: Potoka
authors.affiliation: The Centre of Excellence in Cell-Extracellular Matrix Research, Oulu, Finland.
authors.forename: Seppo
authors.initials: S
authors.lastname: Vainio
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Pediatric General and Thoracic Surgery, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, and Rangos Research Center, Pittsburgh, Pennsylvania; and.
authors.forename: George K
authors.initials: GK
authors.lastname: Gittes
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Nephrology, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania;
authors.forename: Carlton M
authors.initials: CM
authors.lastname: Bates
authors.affiliation: Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania; Division of Nephrology, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; simslucass@upmc.edu.
authors.forename: Sunder
authors.initials: S
authors.lastname: Sims-Lucas
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D005260:Female; D049452:Green Fluorescent Proteins; D007668:Kidney / Q000098:blood supply* / Q000196:embryology / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D009399:Nephrons / Q000166:cytology* / Q000187:drug effects; D009924:Organ Culture Techniques; D010100:Oxygen / Q000378:metabolism / Q000494:pharmacology*; D010477:Perfusion; D037121:Plant Lectins; D012039:Regional Blood Flow / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects
chemical_list: D037121:Plant Lectins; D049452:Green Fluorescent Proteins; D010100:Oxygen
keywords: blood flow; endothelium; hypoxia; kidney development; vasculature