Brain Structure and Function

, Volume 223, Issue 4, pp 1971–1987 | Cite as

Vascular pattern of the dentate gyrus is regulated by neural progenitors

  • Ana Pombero
  • Raquel Garcia-Lopez
  • Alicia Estirado
  • Salvador Martinez
Original Article


Neurogenesis is a vital process that begins during early embryonic development and continues until adulthood, though in the latter case, it is restricted to the subventricular zone and the subgranular zone of the dentate gyrus (DG). In particular, the DG’s neurogenic properties are structurally and functionally unique, which may be related to its singular vascular pattern. Neurogenesis and angiogenesis share molecular signals and act synergistically, supporting the concept of a neurogenic niche as a functional unit between neural precursors cells and their environment, in which the blood vessels play an important role. Whereas it is well known that vascular development controls neural proliferation in the embryonary and in the adult brain, by releasing neurotrophic factors; the potential influence of neural cells on vascular components during angiogenesis is largely unknown. We have demonstrated that the reduction of neural progenitors leads to a significant impairment of vascular development. Since VEGF is a potential regulator in the neurogenesis–angiogenesis crosstalk, we were interested in assessing the possible role of this molecule in the hippocampal neurovascular development. Our results showed that VEGF is the molecule involved in the regulation of vascular development by neural progenitor cells in the DG.


Blood vessel development Dentate gyrus FGFR1 VEGF. 



We thank Dr. A. Barco for providing us Nes-Cre mouse strain. This work was supported by Spanish State Research Agency, through the “Severo Ochoa” Program for Centers of Excellence in R&D (ref. SEV- 2013-0317), by Economy and Competitivity Ministry through Fondos FEDER (SAF2014-59347-C2-1-R), by Generalitat Valenciana Prometeo II Grant (2014/014), by Instituto de Salud Carlos III (RD16/001/0010) (Co-funded by European Regional Development Fund/European Social Fund) and Todos con Natalia Niemann Pick C Association (2016/00084/001).

Compliance with ethical standards


This work was supported by Spanish State Research Agency, through the “Severo Ochoa” Program for Centers of Excellence in R&D (ref. SEV- 2013 − 0317), by Economy and Competitivity Ministry through Fondos FEDER (SAF2014-59347-C2-1-R), by Generalitat Valenciana Prometeo II Grant (2014/014), by Instituto de Salud Carlos III (RD16/001/0010) (Co-funded by European Regional Development Fund/European Social Fund), and Todos con Natalia Niemann Pick C Association (2016/00084/001)

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All animal experimental assays were performed in compliance with the Spanish and European Union laws on animal care in experimentation (Council Directive 86/609/EEC) and approved by the Animal Experimentation Ethics Committee of our University. This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

429_2017_1603_MOESM1_ESM.tif (43.9 mb)
Supplementary Figure 1. Analysis of cre activity in the cortex and the hippocampus. CD13 (green color) was used as a pericyte marker together with RFP (red color) used as a reporter of cre activity. No double labeling was detected in double immunohistochemistry for CD13/RFP in Cx (A-C), DMS (D-F), CA (G-I), and DG (J-L). Scale bar is 25 µm (TIF 44947 KB)


  1. Alon T, Hemo I, Itin A et al (1995) Vascular endothelial growth factor acts as a survival factor for newly formed retinal vessels and has implications for retinopathy of prematurity. Nat Med 1:1024–1028CrossRefPubMedGoogle Scholar
  2. Asai T, Wanaka A, Kato H et al (1993) Differential expression of two members of FGF receptor gene family, FGFR-1 and FGFR-2 mRNA, in the adult rat central nervous system. Brain Res Mol Brain Res 17:174–178CrossRefPubMedGoogle Scholar
  3. Baffert F, Le T, Sennino B et al (2006) Cellular changes in normal blood capillaries undergoing regression after inhibition of VEGF signaling. Am J Physiol Heart Circ Physiol 290:H547–H559CrossRefPubMedGoogle Scholar
  4. Benjamin LE, Golijanin D, Itin A et al (1999) Selective ablation of immature blood vessels in established human tumors follows vascular endothelial growth factor withdrawal. J Clin Invest 103:159–165CrossRefPubMedPubMedCentralGoogle Scholar
  5. Boström M, Hellström Erkenstam N, Kaluza D et al (2014) The hippocampal neurovascular niche during normal development and after irradiation to the juvenile mouse brain. Int J Radiat Biol 90:778–789CrossRefPubMedGoogle Scholar
  6. Breier G, Albrecht U, Sterrer S, Risau W (1992) Expression of vascular endothelial growth factor during embryonic angiogenesis and endothelial cell differentiation. Development 114:521–532PubMedGoogle Scholar
  7. Brown WR (2010) A review of string vessels or collapsed, empty basement membrane tubes. J Alzheimers Dis 21:725–739CrossRefPubMedPubMedCentralGoogle Scholar
  8. Capela A, Temple S (2002) LeX/ssea-1 is expressed by adult mouse CNS stem cells, identifying them as nonependymal. Neuron 35:865–875CrossRefPubMedGoogle Scholar
  9. Chen Q, Jiang L, Li C et al (2012) Haemodynamics-driven developmental pruning of brain vasculature in zebrafish. PLoS Biol 10:e1001374CrossRefPubMedPubMedCentralGoogle Scholar
  10. Corada M, Nyqvist D, Orsenigo F et al (2010) The Wnt/beta-catenin pathway modulates vascular remodeling and specification by upregulating Dll4/Notch signaling. Dev Cell 18:938–949CrossRefPubMedGoogle Scholar
  11. Craig CG, D’sa R, Morshead CM et al (1999) Migrational analysis of the constitutively proliferating subependyma population in adult mouse forebrain. Neuroscience 93:1197–1206CrossRefPubMedGoogle Scholar
  12. Daneman R, Agalliu D, Zhou L et al (2009) Wnt/ -catenin signaling is required for CNS, but not non-CNS, angiogenesis. Proc Natl Acad Sci 106:641–646CrossRefPubMedPubMedCentralGoogle Scholar
  13. Dvorak H (2006) Discovery of vascular permeability factor (VPF). Exp Cell Res 312:522–526Google Scholar
  14. Fabel K, Fabel K, Tam B et al (2003) VEGF is necessary for exercise-induced adult hippocampal neurogenesis. Eur J Neurosci 18:2803–2812CrossRefPubMedGoogle Scholar
  15. Filippov V, Kronenberg G, Pivneva T et al (2003) Subpopulation of nestin-expressing progenitor cells in the adult murine hippocampus shows electrophysiological and morphological characteristics of astrocytes. Mol Cell Neurosci 23:373–382CrossRefPubMedGoogle Scholar
  16. Flamme I, Breier G, Risau W (1995) Vascular endothelial growth factor (VEGF) and VEGF receptor 2 (flk-1) are expressed during vasculogenesis and vascular differentiation in the quail embryo. Dev Biol 169:699–712CrossRefPubMedGoogle Scholar
  17. Franco CA, Jones ML, Bernabeu MO et al (2015) Correction: dynamic endothelial cell rearrangements drive developmental vessel regression. PLoS Biol 13:e1002163CrossRefPubMedPubMedCentralGoogle Scholar
  18. Fuentealba LC, Obernier K, Alvarez-Buylla A (2012) Adult neural stem cells bridge their niche. Cell Stem Cell 10:698–708CrossRefPubMedPubMedCentralGoogle Scholar
  19. Gerhardt H, Golding M, Fruttiger M et al (2003) VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia. J Cell Biol 161:1163–1177CrossRefPubMedPubMedCentralGoogle Scholar
  20. Gerhardt H, Ruhrberg C, Abramsson A et al (2004) Neuropilin-1 is required for endothelial tip cell guidance in the developing central nervous system. Dev Dyn 231:503–509CrossRefPubMedGoogle Scholar
  21. Goldman SA, Chen Z (2011) Perivascular instruction of cell genesis and fate in the adult brain. Nat Neurosci 14:1382–1389CrossRefPubMedPubMedCentralGoogle Scholar
  22. Gould E (1999) Neurogenesis in the Neocortex of Adult Primates. Science 286:548–552CrossRefPubMedGoogle Scholar
  23. Gould E, Reeves AJ, Fallah M et al (1999) Hippocampal neurogenesis in adult Old World primates. Proc Natl Acad Sci 96:5263–5267CrossRefPubMedPubMedCentralGoogle Scholar
  24. Graus-Porta D, Blaess S, Senften M et al (2001) Beta1-class integrins regulate the development of laminae and folia in the cerebral and cerebellar cortex. Neuron 31:367–379CrossRefPubMedGoogle Scholar
  25. Groppa E, Brkic S, Bovo E et al (2015) VEGF dose regulates vascular stabilization through Semaphorin3A and the Neuropilin-1 + monocyte/TGF-β1 paracrine axis. EMBO Mol Med 7:1366–1384CrossRefPubMedPubMedCentralGoogle Scholar
  26. Gu C, Yoshida Y, Livet J et al (2005) Semaphorin 3E and plexin-D1 control vascular pattern independently of neuropilins. Science 307:265–268CrossRefPubMedGoogle Scholar
  27. Heuchan AM, Evans N, Henderson-Smart DJ, Simpson JM (2002) Perinatal risk factors for major intraventricular haemorrhage in the australian and new zealand noenatal network, 1005–97. Arch Dis Child Fetal Neoanatal Ed 86:F86–F90CrossRefGoogle Scholar
  28. Hofmann G, Balgooyen L, Mattis J et al (2016) Hilar somatostatin interneuron loss reduces dentate gyrus inhibition in a mouse model of temporal lobe epilepsy. Epilepsia 57:977–983CrossRefPubMedPubMedCentralGoogle Scholar
  29. Inai T, Mancuso M, Hashizume H et al (2004) Inhibition of vascular endothelial growth factor (VEGF) signaling in cancer causes loss of endothelial fenestrations, regression of tumor vessels, and appearance of basement membrane ghosts. Am J Pathol 165:35–52CrossRefPubMedPubMedCentralGoogle Scholar
  30. Jakeman LB, Armanini M, Phillips HS, Ferrara N (1993) Developmental expression of binding sites and messenger ribonucleic acid for vascular endothelial growth factor suggests a role for this protein in vasculogenesis and angiogenesis. Endocrinology 133:848–859CrossRefPubMedGoogle Scholar
  31. Jiao Q, Xie W-L, Wang Y-Y et al (2013) Spatial relationship between NSCs/NPCs and microvessels in rat brain along prenatal and postnatal development. Int J Dev Neurosci 31:280–285CrossRefPubMedGoogle Scholar
  32. Jin K, Zhu Y, Sun Y et al (2002) Vascular endothelial growth factor (VEGF) stimulates neurogenesis in vitro and in vivo. Proc Natl Acad Sci USA 99:11946–11950CrossRefPubMedPubMedCentralGoogle Scholar
  33. Kang W, Hebert JM (2015) FGF Signaling Is Necessary for Neurogenesis in Young Mice and Sufficient to Reverse Its Decline in Old Mice. J Neurosci 35:10217–10223CrossRefPubMedPubMedCentralGoogle Scholar
  34. Koch S, Claesson-Welsh L (2012) Signal transduction by vascular endothelial growth factor receptors. Cold Spring Harb Perspect Med 2:a006502CrossRefPubMedPubMedCentralGoogle Scholar
  35. Kokovay E, Goderie S, Wang Y et al (2010) Adult SVZ lineage cells home to and leave the vascular niche via differential responses to SDF1/CXCR4 signaling. Cell Stem Cell 7:163–173CrossRefPubMedPubMedCentralGoogle Scholar
  36. Korn C, Augustin HG (2015) Mechanisms of vessel pruning and regression. Dev Cell 34:5–17CrossRefPubMedGoogle Scholar
  37. Korn C, Scholz B, Hu J et al (2014) Endothelial cell-derived non-canonical Wnt ligands control vascular pruning in angiogenesis. Development 141:1757–1766CrossRefPubMedGoogle Scholar
  38. Kuhn HG, Dickinson-Anson H, Gage FH (1996) Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation. J Neurosci 16:2027–2033CrossRefPubMedGoogle Scholar
  39. Leventhal C, Rafii S, Rafii D et al (1999) Endothelial trophic support of neuronal production and recruitment from the adult mammalian subependyma. Mol Cell Neurosci 13:450–464CrossRefPubMedGoogle Scholar
  40. Liang H, Hippenmeyer S, Ghashghaei HT (2012) A Nestin-cre transgenic mouse is insufficient for recombination in early embryonic neural progenitors. Biol Open 1:1200–1203CrossRefPubMedPubMedCentralGoogle Scholar
  41. Licht T, Keshet E (2015) The vascular niche in adult neurogenesis. Mech Dev 138 Pt 1:56–62CrossRefGoogle Scholar
  42. Lobov IB, Rao S, Carroll TJ et al (2005) WNT7b mediates macrophage-induced programmed cell death in patterning of the vasculature. Nature 437:417–421CrossRefPubMedPubMedCentralGoogle Scholar
  43. Louissaint A Jr, Rao S, Leventhal C, Goldman SA (2002) Coordinated interaction of neurogenesis and angiogenesis in the adult songbird brain. Neuron 34:945–960CrossRefPubMedGoogle Scholar
  44. Lu X, Le Noble F, Yuan L et al (2004) The netrin receptor UNC5B mediates guidance events controlling morphogenesis of the vascular system. Nature 432:179–186CrossRefPubMedGoogle Scholar
  45. Luskin MB, Scott Boone M (1994) Rate and pattern of migration of lineally-related olfactory bulb interneurons generated postnatally in the subventricular zone of the rat. Chem Senses 19:695–714CrossRefPubMedGoogle Scholar
  46. Ma S, Huang Z (2015) Neural regulation of CNS angiogenesis during development. Front Biol 10:61–73CrossRefGoogle Scholar
  47. Ma S, Kwon HJ, Johng H et al (2013) Radial glial neural progenitors regulate nascent brain vascular network stabilization via inhibition of Wnt signaling. PLoS Biol 11:e1001469CrossRefPubMedPubMedCentralGoogle Scholar
  48. Matsuzaki H (2000) VEGF rescues hippocampal neurons from glutamate-induced toxicity through Akt and ERK activation. Neurosci Res 38:S51CrossRefGoogle Scholar
  49. Matsuzaki H, Tamatani M, Yamaguchi A et al (2001) Vascular endothelial growth factor rescues hippocampal neurons from glutamate-induced toxicity: signal transduction cascades. FASEB J 15:1218–1220CrossRefPubMedGoogle Scholar
  50. Meadows KN, Bryant P, Pumiglia K (2001) Vascular endothelial growth factor induction of the angiogenic phenotype requires Ras activation. J Biol Chem 276:49289–49298CrossRefPubMedGoogle Scholar
  51. Meeson AP, Argilla M, Ko K et al (1999) VEGF deprivation-induced apoptosis is a component of programmed capillary regression. Development 126:1407–1415PubMedGoogle Scholar
  52. Monje ML, Mizumatsu S, Fike JR, Palmer TD (2002) Irradiation induces neural precursor-cell dysfunction. Nat Med 8:955–962CrossRefPubMedGoogle Scholar
  53. Ng Y-S, Rohan R, Sunday ME et al (2001) Differential expression of VEGF isoforms in mouse during development and in the adult. Dev Dyn 220:112–121Google Scholar
  54. Ohkubo Y, Uchida AO, Shin D et al (2004) Fibroblast growth factor receptor 1 is required for the proliferation of hippocampal progenitor cells and for hippocampal growth in mouse. J Neurosci 24:6057–6069CrossRefPubMedGoogle Scholar
  55. Palmer TD, Willhoite AR, Gage FH (2000) Vascular niche for adult hippocampal neurogenesis. J Comp Neurol 425:479–494CrossRefPubMedGoogle Scholar
  56. Phng L-K, Potente M, Leslie JD et al (2009) Nrarp coordinates endothelial Notch and Wnt signaling to control vessel density in angiogenesis. Dev Cell 16:70–82CrossRefPubMedGoogle Scholar
  57. Plate KH, Breier G, Farrell CL, Risau W (1992) Platelet-derived growth factor receptor-beta is induced during tumor development and upregulated during tumor progression in endothelial cells in human gliomas. Lab Invest 67:529–534PubMedGoogle Scholar
  58. Pombero A, Garcia-Lopez R, Martinez S (2016) Brain mesenchymal stem cells: physiology and pathological implications. Dev Growth Differ 58:469–480CrossRefPubMedGoogle Scholar
  59. Proctor JM, Zang K, Wang D et al (2005) Vascular development of the brain requires beta8 integrin expression in the neuroepithelium. J Neurosci 25:9940–9948CrossRefPubMedPubMedCentralGoogle Scholar
  60. Qian X, Shen Q, Goderie SK et al (2000) Timing of CNS Cell Generation. Neuron 28:69–80CrossRefPubMedGoogle Scholar
  61. Raab S, Beck H, Gaumann A et al (2004) Impaired brain angiogenesis and neuronal apoptosis induced by conditional homozygous inactivation of vascular endothelial growth factor. Thromb Haemost 91:595–605PubMedGoogle Scholar
  62. Rao S, Lobov IB, Vallance JE et al (2007) Obligatory participation of macrophages in an angiopoietin 2-mediated cell death switch. Development 134:4449–4458CrossRefPubMedPubMedCentralGoogle Scholar
  63. Reichardt LF (2006) Neurotrophin-regulated signalling pathways. Philos Trans R Soc Lond B Biol Sci 361:1545–1564CrossRefPubMedPubMedCentralGoogle Scholar
  64. Rickmann M, Amaral DG, Cowan WM (1987) Organization of radial glial cells during the development of the rat dentate gyrus. J Comp Neurol 264:449–479CrossRefPubMedGoogle Scholar
  65. Riquelme PA, Drapeau E, Doetsch F (2008) Brain micro-ecologies: neural stem cell niches in the adult mammalian brain. Philos Trans R Soc Lond B Biol Sci 363:123–137CrossRefPubMedGoogle Scholar
  66. Ruhrberg C (2003) Growing and shaping the vascular tree: multiple roles for VEGF. BioEssays 25:1052–1060CrossRefPubMedGoogle Scholar
  67. Ruhrberg C, Gerhardt H, Golding M et al (2002) Spatially restricted patterning cues provided by heparin-binding VEGF-A control blood vessel branching morphogenesis. Genes Dev 16:2684–2698CrossRefPubMedPubMedCentralGoogle Scholar
  68. Scott A, Powner MB, Gandhi P et al (2010) Astrocyte-derived vascular endothelial growth factor stabilizes vessels in the developing retinal vasculature. PLoS One 5:e11863CrossRefPubMedPubMedCentralGoogle Scholar
  69. Shen Q, Goderie SK, Jin L et al (2004) Endothelial cells stimulate self-renewal and expand neurogenesis of neural stem cells. Science 304:1338–1340CrossRefPubMedGoogle Scholar
  70. Siebzehnrubl FA, Blumcke I (2008) Neurogenesis in the human hippocampus and its relevance to temporal lobe epilepsies. Epilepsia 49(Suppl 5):55–65CrossRefPubMedGoogle Scholar
  71. Sondell M, Lundborg G, Kanje M (1999) Vascular endothelial growth factor has neurotrophic activity and stimulates axonal outgrowth, enhancing cell survival and Schwann cell proliferation in the peripheral nervous system. J Neurosci 19:5731–5740PubMedGoogle Scholar
  72. Stenman JM, Rajagopal J, Carroll TJ et al (2008) Canonical Wnt signaling regulates organ-specific assembly and differentiation of CNS vasculature. Science 322:1247–1250CrossRefPubMedGoogle Scholar
  73. Stolp HB, Molnár Z (2015) Neurogenic niches in the brain: help and hindrance of the barrier systems. Front Neurosci 9:20CrossRefPubMedPubMedCentralGoogle Scholar
  74. Stubbs D, DeProto J, Nie K et al (2009) Neurovascular congruence during cerebral cortical development. Cereb Cortex 19:i32–i41CrossRefPubMedPubMedCentralGoogle Scholar
  75. Thakker GD, Hajjar DP, Muller WA, Rosengart TK (1999) The role of phosphatidylinositol 3-kinase in vascular endothelial growth factor signaling. J Biol Chem 274:10002–10007CrossRefPubMedGoogle Scholar
  76. Tortora D, Veverino M, Malova M et al (2018) Differences in subependymal vein anatomy may predispose preterm infants to GMH-IVH. Arch Dis Child Fetal Neonatal Ed 103:F59–F65CrossRefPubMedGoogle Scholar
  77. Tronche F, Kellendonk C, Kretz O et al (1999) Disruption of the glucocorticoid receptor gene in the nervous system results in reduced anxiety. Nat Genet 23:99–103CrossRefPubMedGoogle Scholar
  78. Udan RS, Vadakkan TJ, Dickinson ME (2013) Dynamic responses of endothelial cells to changes in blood flow during vascular remodeling of the mouse yolk sac. Development 140:4041–4050CrossRefPubMedPubMedCentralGoogle Scholar
  79. Urbán N, Guillemot F (2014) Neurogenesis in the embryonic and adult brain: same regulators, different roles. Front Cell Neurosci 8:396CrossRefPubMedPubMedCentralGoogle Scholar
  80. Wagner JP, Black IB, DiCicco-Bloom E (1999) Stimulation of neonatal and adult brain neurogenesis by subcutaneous injection of basic fibroblast growth factor. J Neurosci 19:6006–6016PubMedGoogle Scholar
  81. Weickert CS, Kittell DA, Saunders RC et al (2005) Basic fibroblast growth factor and fibroblast growth factor receptor-1 in the human hippocampal formation. Neuroscience 131:219–233CrossRefPubMedGoogle Scholar
  82. Wu LW, Mayo LD, Dunbar JD et al (2000) VRAP is an adaptor protein that binds KDR, a receptor for vascular endothelial cell growth factor. J Biol Chem 275:6059–6062CrossRefPubMedGoogle Scholar
  83. Xiao Z, Kong Y, Yang S et al (2007) Upregulation of Flk-1 by bFGF via the ERK pathway is essential for VEGF-mediated promotion of neural stem cell proliferation. Cell Res 17:73–79CrossRefPubMedGoogle Scholar
  84. Xu X, Qiao W, Li C, Deng C (2002) Generation of Fgfr1 conditional knockout mice. Genesis 32:85–86CrossRefPubMedGoogle Scholar
  85. Yamashima T, Tonchev AB, Vachkov IH et al (2004) Vascular adventitia generates neuronal progenitors in the monkey hippocampus after ischemia. Hippocampus 14:861–875CrossRefPubMedGoogle Scholar
  86. Yancopoulos GD, Davis S, Gale NW et al (2000) Vascular-specific growth factors and blood vessel formation. Nature 407:242–248CrossRefPubMedGoogle Scholar
  87. Zachary I (2001) Signaling mechanisms mediating vascular protective actions of vascular endothelial growth factor. Am J Physiol Cell Physiol 280:C1375–C1386CrossRefPubMedGoogle Scholar
  88. Zhao C, Deng W, Gage FH (2008) Mechanisms and functional implications of adult neurogenesis. Cell 132:645–660CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Instituto de Neurociencias, UMH-CSICAlicanteSpain
  2. 2.IMIB-Arrixaca, University of MurciaMurciaSpain
  3. 3.Centro de Investigación Biomédica En Red en Salud Mental (CIBERSAM)MadridSpain

Personalised recommendations