Abstract
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.
Similar content being viewed by others
References
Cleaver O, Tonissen KF, Saha MS, Krieg PA (1997) Neovascularization of the Xenopus embryo. Dev Dyn 210(1):66–77
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
Coultas L, Nieuwenhuis E, Anderson GA, Cabezas J, Nagy A, Henkelman RM, Hui CC, Rossant J (2010) Hedgehog regulates distinct vascular patterning events through VEGF-dependent and -independent mechanisms. Blood 116(4):653–660. https://doi.org/10.1182/blood-2009-12-256644
Drake CJ, Fleming PA (2000) Vasculogenesis in the day 6.5 to 9.5 mouse embryo. Blood 95(5):1671–1679
Azizoglu DB, Chong DC, Villasenor A, Magenheim J, Barry DM, Lee S, Marty-Santos L, Fu S, Dor Y, Cleaver O (2016) Vascular development in the vertebrate pancreas. Dev Biol 420(1):67–78. https://doi.org/10.1016/j.ydbio.2016.10.009
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
O’Brien LL, McMahon AP (2014) Induction and patterning of the metanephric nephron. Semin Cell Dev Biol 36:31–38. https://doi.org/10.1016/j.semcdb.2014.08.014
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
Gao L, Yang Z, Hiremath C, Zimmerman SE, Long B, Brakeman PR, Mostov KE, Bryant DM, Luby-Phelps K, Marciano DK (2017) Afadin orients cell division to position the tubule lumen in developing renal tubules. Development 144(19):3511–3520. https://doi.org/10.1242/dev.148908
Robert B, St John PL, Abrahamson DR (1998) Direct visualization of renal vascular morphogenesis in Flk1 heterozygous mutant mice. Am J Physiol 275(1 Pt 2):F164–F172
Tufro A, Norwood VF, Carey RM, Gomez RA (1999) Vascular endothelial growth factor induces nephrogenesis and vasculogenesis. J Am Soc Nephrol 10(10):2125–2134
Munro DAD, Hohenstein P, Davies JA (2017) Cycles of vascular plexus formation within the nephrogenic zone of the developing mouse kidney. Sci Rep 7(1):3273. https://doi.org/10.1038/s41598-017-03808-4
Vaughan MR, Quaggin SE (2008) How do mesangial and endothelial cells form the glomerular tuft? J Am Soc Nephrol 19(1):24–33. https://doi.org/10.1681/ASN.2007040471
Gao X, Chen X, Taglienti M, Rumballe B, Little MH, Kreidberg JA (2005) Angioblast-mesenchyme induction of early kidney development is mediated by Wt1 and Vegfa. Development 132(24):5437–5449. https://doi.org/10.1242/dev.02095
Munro DAD, Hohenstein P, Coate TM, Davies JA (2017) Refuting the hypothesis that semaphorin-3f/neuropilin-2 exclude blood vessels from the cap mesenchyme in the developing kidney. Dev Dyn 246(12):1047–1056. https://doi.org/10.1002/dvdy.24592
Hurtado R, Zewdu R, Mtui J, Liang C, Aho R, Kurylo C, Selleri L, Herzlinger D (2015) Pbx1-dependent control of VMC differentiation kinetics underlies gross renal vascular patterning. Development 142(15):2653–2664. https://doi.org/10.1242/dev.124776
Villasenor A, Chong DC, Cleaver O (2008) Biphasic Ngn3 expression in the developing pancreas. Dev Dyn 237(11):3270–3279. https://doi.org/10.1002/dvdy.21740
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
Patro R, Duggal G, Love MI, Irizarry RA, Kingsford C (2017) Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods 14(4):417–419. https://doi.org/10.1038/nmeth.4197
Soneson C, Love MI, Robinson MD (2015) Differential analyses for RNA-seq: transcript-level estimates improve gene-level inferences. F1000Res 4:1521. https://doi.org/10.12688/f1000research.7563.2
Love MI, Huber W, Anders S (2014) Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15(12):550. https://doi.org/10.1186/s13059-014-0550-8
Langfelder P, Horvath S (2008) WGCNA: an R package for weighted correlation network analysis. BMC Bioinform 9:559. https://doi.org/10.1186/1471-2105-9-559
Ema M, Takahashi S, Rossant J (2006) Deletion of the selection cassette, but not cis-acting elements, in targeted Flk1-lacZ allele reveals Flk1 expression in multipotent mesodermal progenitors. Blood 107(1):111–117. https://doi.org/10.1182/blood-2005-05-1970
Shalaby F, Rossant J, Yamaguchi TP, Gertsenstein M, Wu XF, Breitman ML, Schuh AC (1995) Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature 376(6535):62–66
Moyon D, Pardanaud L, Yuan L, Breant C, Eichmann A (2001) Plasticity of endothelial cells during arterial-venous differentiation in the avian embryo. Development 128(17):3359–3370
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
Chong DC, Koo Y, Xu K, Fu S, Cleaver O (2011) Stepwise arteriovenous fate acquisition during mammalian vasculogenesis. Dev Dyn 240(9):2153–2165. https://doi.org/10.1002/dvdy.22706
Klagsbrun M, Takashima S, Mamluk R (2002) The role of neuropilin in vascular and tumor biology. Adv Exp Med Biol 515:33–48
dela Paz NG, dela D’Amore PA (2009) Arterial versus venous endothelial cells. Cell Tissue Res 335(1):5–16. https://doi.org/10.1007/s00441-008-0706-5
Villasenor A, Wang ZV, Rivera LB, Ocal O, Asterholm IW, Scherer PE, Brekken RA, Cleaver O, Wilkie TM (2010) Rgs16 and Rgs8 in embryonic endocrine pancreas and mouse models of diabetes. Dis Model Mech 3(9–10):567–580. https://doi.org/10.1242/dmm.003210
Magenheim J, Ilovich O, Lazarus A, Klochendler A, Ziv O, Werman R, Hija A, Cleaver O, Mishani E, Keshet E, Dor Y (2011) Blood vessels restrain pancreas branching, differentiation and growth. Development 138(21):4743–4752. https://doi.org/10.1242/dev.066548
Zeng X, Wert SE, Federici R, Peters KG, Whitsett JA (1998) VEGF enhances pulmonary vasculogenesis and disrupts lung morphogenesis in vivo. Dev Dyn 211(3):215–227
Lazarus A, Del-Moral PM, Ilovich O, Mishani E, Warburton D, Keshet E (2011) A perfusion-independent role of blood vessels in determining branching stereotypy of lung airways. Development 138(11):2359–2368. https://doi.org/10.1242/dev.060723
Costantini F, Kopan R (2010) Patterning a complex organ: branching morphogenesis and nephron segmentation in kidney development. Dev Cell 18(5):698–712. https://doi.org/10.1016/j.devcel.2010.04.008
Eremina V, Sood M, Haigh J, Nagy A, Lajoie G, Ferrara N, Gerber HP, Kikkawa Y, Miner JH, Quaggin SE (2003) Glomerular-specific alterations of VEGF-A expression lead to distinct congenital and acquired renal diseases. J Clin Invest 111(5):707–716
Kitamoto Y, Tokunaga H, Tomita K (1997) Vascular endothelial growth factor is an essential molecule for mouse kidney development: glomerulogenesis and nephrogenesis. J Clin Invest 99(10):2351–2357
Lindstrom NO, Tran T, Guo J, Rutledge E, Parvez RK, Thornton ME, Grubbs B, McMahon JA, McMahon AP (2018) Conserved and divergent molecular and anatomic features of human and mouse nephron patterning. J Am Soc Nephrol. https://doi.org/10.1681/ASN.2017091036
Yang Z, Zimmerman SE, Tsunezumi J, Braitsch C, Trent C, Bryant DM, Cleaver O, Gonzalez-Manchon C, Marciano DK (2016) Role of CD34 family members in lumen formation in the developing kidney. Dev Biol 418(1):66–74. https://doi.org/10.1016/j.ydbio.2016.08.009
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
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
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
Hum S, Rymer C, Schaefer C, Bushnell D, Sims-Lucas S (2014) Ablation of the renal stroma defines its critical role in nephron progenitor and vasculature patterning. PLoS ONE 9(2):e88400. https://doi.org/10.1371/journal.pone.0088400
Sequeira-Lopez ML, Lin EE, Li M, Hu Y, Sigmund CD, Gomez RA (2015) The earliest metanephric arteriolar progenitors and their role in kidney vascular development. Am J Physiol Regul Integr Comp Physiol 308(2):R138–R149. https://doi.org/10.1152/ajpregu.00428.2014
Nordsletten DA, Blackett S, Bentley MD, Ritman EL, Smith NP (2006) Structural morphology of renal vasculature. Am J Physiol Heart Circ Physiol 291(1):H296–H309. https://doi.org/10.1152/ajpheart.00814.2005
Tufro A (2000) VEGF spatially directs angiogenesis during metanephric development in vitro. Dev Biol 227(2):558–566
Abrahamson DR (2009) Development of kidney glomerular endothelial cells and their role in basement membrane assembly. Organogenesis 5(1):275–287
Mazumdar J, O’Brien WT, Johnson RS, LaManna JC, Chavez JC, Klein PS, Simon MC (2010) O2 regulates stem cells through Wnt/beta-catenin signalling. Nat Cell Biol 12(10):1007–1013. https://doi.org/10.1038/ncb2102
Culver JC, Vadakkan TJ, Dickinson ME (2013) A specialized microvascular domain in the mouse neural stem cell niche. PLoS ONE 8(1):e53546. https://doi.org/10.1371/journal.pone.0053546
Takubo K, Goda N, Yamada W, Iriuchishima H, Ikeda E, Kubota Y, Shima H, Johnson RS, Hirao A, Suematsu M, Suda T (2010) Regulation of the HIF-1alpha level is essential for hematopoietic stem cells. Cell Stem Cell 7(3):391–402. https://doi.org/10.1016/j.stem.2010.06.020
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
Saxén L (1987) Vascularization of the nephron. In: Barlow PW, Green PB, Wylie CC (eds) Organogenesis of the kidney. Cambridge University Press, Cambridge, pp 129–142
Molema G, Aird WC (2012) Vascular heterogeneity in the kidney. Semin Nephrol 32(2):145–155. https://doi.org/10.1016/j.semnephrol.2012.02.001
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
Aird WC (2012) Endothelial cell heterogeneity. Cold Spring Harb Perspect Med 2(1):a006429. https://doi.org/10.1101/cshperspect.a006429
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
Brunskill EW, Potter SS (2010) Gene expression programs of mouse endothelial cells in kidney development and disease. PLoS ONE 5(8):e12034. https://doi.org/10.1371/journal.pone.0012034
Acknowledgements
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).
Author contributions
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.
Funding
This work was supported by National Institute of Diabetes and Digestive and Kidney Diseases DK099478 to DKM., DK080004, DK095057, DK106743 to TJC and DK106743, DK079862 to OC; CPRIT RP110405; and National Institute of Heart, Lung, and Blood HL113498 to OC. Deposited in PMC for release after 12 months.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing or financial interests.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Daniel, E., Azizoglu, D.B., Ryan, A.R. et al. Spatiotemporal heterogeneity and patterning of developing renal blood vessels. Angiogenesis 21, 617–634 (2018). https://doi.org/10.1007/s10456-018-9612-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10456-018-9612-y