Spatiotemporal heterogeneity and patterning of developing renal blood vessels
The kidney vasculature facilitates the excretion of wastes, the dissemination of hormones, and the regulation of blood chemistry. To carry out these diverse functions, the vasculature is regionalized within the kidney and along the nephron. However, when and how endothelial regionalization occurs remains unknown. Here, we examine the developing kidney vasculature to assess its 3-dimensional structure and transcriptional heterogeneity. First, we observe that endothelial cells (ECs) grow coordinately with the kidney bud as early as E10.5, and begin to show signs of specification by E13.5 when the first arteries can be identified. We then focus on how ECs pattern and remodel with respect to the developing nephron and collecting duct epithelia. ECs circumscribe nephron progenitor populations at the distal tips of the ureteric bud (UB) tree and form stereotyped cruciform structures around each tip. Beginning at the renal vesicle (RV) stage, ECs form a continuous plexus around developing nephrons. The endothelial plexus envelops and elaborates with the maturing nephron, becoming preferentially enriched along the early distal tubule. Lastly, we perform transcriptional and immunofluorescent screens to characterize spatiotemporal heterogeneity in the kidney vasculature and identify novel regionally enriched genes. A better understanding of development of the kidney vasculature will help instruct engineering of properly vascularized ex vivo kidneys and evaluate diseased kidneys.
KeywordsEndothelium Epithelium Blood vessel Nephron Vascular patterning Endothelial cell heterogeneity
We thank Janet Rossant for the Flk1-eGFP mouse line, as well as members of the Cleaver lab, including Caitlin Braitsch, Xiaowu Gu, and David Barry, for discussions and critical reading of the manuscript. We thank the Genepaint.org database for in situ hybridization data (where noted).
Experiments were performed by ED, DBA, ARR, TAW, CC, and GISTJC, DKM, and OC supervised the project and contributed to analysis. ED and OC wrote the text of this article with input from co-authors.
Compliance with ethical standards
Conflict of interest
The authors declare no competing or financial interests.
- 2.Herbert SP, Huisken J, Kim TN, Feldman ME, Houseman BT, Wang RA, Shokat KM, Stainier DY (2009) Arterial-venous segregation by selective cell sprouting: an alternative mode of blood vessel formation. Science 326(5950):294–298. https://doi.org/10.1126/science.1178577 CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Oxburgh L, Carroll TJ, Cleaver O, Gossett DR, Hoshizaki DK, Hubbell JA, Humphreys BD, Jain S, Jensen J, Kaplan DL, Kesselman C, Ketchum CJ, Little MH, McMahon AP, Shankland SJ, Spence JR, Valerius MT, Wertheim JA, Wessely O, Zheng Y, Drummond IA (2017) (Re)building a kidney. J Am Soc Nephrol 28(5):1370–1378. https://doi.org/10.1681/ASN.2016101077 CrossRefPubMedGoogle Scholar
- 8.Yang Z, Zimmerman S, Brakeman PR, Beaudoin GM 3rd, Reichardt LF, Marciano DK (2013) De novo lumen formation and elongation in the developing nephron: a central role for afadin in apical polarity. Development 140(8):1774–1784. https://doi.org/10.1242/dev.087957 CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Acar M, Kocherlakota KS, Murphy MM, Peyer JG, Oguro H, Inra CN, Jaiyeola C, Zhao Z, Luby-Phelps K, Morrison SJ (2015) Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal. Nature 526(7571):126–130. https://doi.org/10.1038/nature15250 CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Rymer C, Paredes J, Halt K, Schaefer C, Wiersch J, Zhang G, Potoka D, Vainio S, Gittes GK, Bates CM, Sims-Lucas S (2014) Renal blood flow and oxygenation drive nephron progenitor differentiation. Am J Physiol Renal Physiol 307(3):F337–F345. https://doi.org/10.1152/ajprenal.00208.2014 CrossRefPubMedPubMedCentralGoogle Scholar
- 39.Lindstrom NO, McMahon JA, Guo J, Tran T, Guo Q, Rutledge E, Parvez RK, Saribekyan G, Schuler RE, Liao C, Kim AD, Abdelhalim A, Ruffins SW, Thornton ME, Basking L, Grubbs B, Kesselman C, McMahon AP (2018) Conserved and divergent features of human and mouse kidney organogenesis. J Am Soc Nephrol. https://doi.org/10.1681/ASN.2017080887 Google Scholar
- 40.Lindstrom NO, Guo J, Kim AD, Tran T, Guo Q, De Sena BG, Ransick A, Parvez RK, Thornton ME, Basking L, Grubbs B, McMahon JA, Smith AD, McMahon AP (2018) Conserved and divergent features of mesenchymal progenitor cell types within the cortical nephrogenic niche of the human and mouse kidney. J Am Soc Nephrol. https://doi.org/10.1681/ASN.2017080890 Google Scholar
- 41.O’Brien LL, Guo Q, Lee Y, Tran T, Benazet JD, Whitney PH, Valouev A, McMahon AP (2016) Differential regulation of mouse and human nephron progenitors by the Six family of transcriptional regulators. Development 143(4):595–608. https://doi.org/10.1242/dev.127175 CrossRefPubMedPubMedCentralGoogle Scholar
- 50.Kimura W, Xiao F, Canseco DC, Muralidhar S, Thet S, Zhang HM, Abderrahman Y, Chen R, Garcia JA, Shelton JM, Richardson JA, Ashour AM, Asaithamby A, Liang H, Xing C, Lu Z, Zhang CC, Sadek HA (2015) Hypoxia fate mapping identifies cycling cardiomyocytes in the adult heart. Nature 523(7559):226–230. https://doi.org/10.1038/nature14582 CrossRefPubMedGoogle Scholar
- 53.Aird WC (2007) Phenotypic heterogeneity of the endothelium: I. Structure, function, and mechanisms. Circ Res 100(2):158–173. https://doi.org/10.1161/01.RES.0000255691.76142.4a CrossRefPubMedGoogle Scholar
- 55.Nolan DJ, Ginsberg M, Israely E, Palikuqi B, Poulos MG, James D, Ding BS, Schachterle W, Liu Y, Rosenwaks Z, Butler JM, Xiang J, Rafii A, Shido K, Rabbany SY, Elemento O, Rafii S (2013) Molecular signatures of tissue-specific microvascular endothelial cell heterogeneity in organ maintenance and regeneration. Dev Cell 26(2):204–219. https://doi.org/10.1016/j.devcel.2013.06.017 CrossRefPubMedGoogle Scholar