Skip to main content

Histological Assessment of Angiogenesis in the Hypoxic Central Nervous System

  • Protocol
Cerebral Angiogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1135))

  • 6955 Accesses

Abstract

Angiogenesis, the sprouting of new capillaries from preexisting vessels, is an integral part of both normal development and numerous pathological conditions such as tumor growth, inflammation, and stroke. The development of angiogenesis assays has been critical in understanding this process in both the context of disease and normal physiology. With the growing availability of antibodies against angiogenic markers as well as advances in microscopy and imaging analysis software, a more comprehensive assessment of the angiogenesis process is beginning to take form (Milner et al., Stroke 39:191–197, 2008; Freitas-Andrade et al., J Cereb Blood Flow Metab 32:663–675, 2012; Li et al., Glia 58:1157–1167, 2010; Dore-Duffy and LaManna, Antioxid Redox Signal 9:1363–1371, 2007). This chapter describes an in vivo method of inducing brain angiogenesis in mice by chronic exposure to mild hypoxia. In addition, a detailed procedure of quantifying angiogenesis using multiple immunofluorescent labeling of mouse brain tissue sections is also presented.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Milner R, Hung S, Wang X, Berg GI, Spatz M, del Zoppo GJ (2008) Responses of endothelial cell and astrocyte matrix-integrin receptors to ischemia mimic those observed in the neurovascular unit. Stroke 39:191–197

    Article  PubMed Central  PubMed  Google Scholar 

  2. Freitas-Andrade M, Carmeliet P, Charlebois C, Stanimirovic DB, Moreno MJ (2012) PlGF knockout delays brain vessel growth and maturation upon systemic hypoxic challenge. J Cereb Blood Flow Metab 32:663–675

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Li L, Welser JV, Dore-Duffy P, del Zoppo GJ, Lamanna JC, Milner R (2010) In the hypoxic central nervous system, endothelial cell proliferation is followed by astrocyte activation, proliferation, and increased expression of the alpha 6 beta 4 integrin and dystroglycan. Glia 58:1157–1167

    PubMed Central  PubMed  Google Scholar 

  4. Dore-Duffy P, LaManna JC (2007) Physiologic angiodynamics in the brain. Antioxid Redox Signal 9:1363–1371

    Article  CAS  PubMed  Google Scholar 

  5. Freitas-Andrade M, Carmeliet P, Stanimirovic DB, Moreno M (2008) VEGFR-2-mediated increased proliferation and survival in response to oxygen and glucose deprivation in PlGF knockout astrocytes. J Neurochem 107:756–767

    Article  CAS  PubMed  Google Scholar 

  6. Stanimirovic DB, Friedman A (2012) Pathophysiology of the neurovascular unit: disease cause or consequence? J Cereb Blood Flow Metab 32:1207–1221

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Beck H, Acker T, Puschel AW, Fujisawa H, Carmeliet P, Plate KH (2002) Cell type-specific expression of neuropilins in an MCA-occlusion model in mice suggests a potential role in post-ischemic brain remodeling. J Neuropathol Exp Neurol 61:339–350

    CAS  PubMed  Google Scholar 

  8. LaManna JC, Chavez JC, Pichiule P (2004) Structural and functional adaptation to hypoxia in the rat brain. J Exp Biol 207:3163–3169

    Article  CAS  PubMed  Google Scholar 

  9. Ratan RR, Siddiq A, Smirnova N, Karpisheva K, Haskew-Layton R, McConoughey S, Langley B, Estevez A, Huerta PT, Volpe B, Roy S, Sen CK, Gazaryan I, Cho S, Fink M, LaManna J (2007) Harnessing hypoxic adaptation to prevent, treat, and repair stroke. J Mol Med (Berl) 85:1331–1338

    Article  Google Scholar 

  10. LaManna JC, Vendel LM, Farrell RM (1992) Brain adaptation to chronic hypobaric hypoxia in rats. J Appl Physiol 72:2238–2243

    CAS  PubMed  Google Scholar 

  11. Xu K, Lamanna JC (2006) Chronic hypoxia and the cerebral circulation. J Appl Physiol 100:725–730

    Article  CAS  PubMed  Google Scholar 

  12. Al Ahmad A, Gassmann M, Ogunshola OO (2009) Maintaining blood-brain barrier integrity: pericytes perform better than astrocytes during prolonged oxygen deprivation. J Cell Physiol 218:612–622

    Article  CAS  PubMed  Google Scholar 

  13. Armulik A, Abramsson A, Betsholtz C (2005) Endothelial/pericyte interactions. Circ Res 97:512–523

    Article  CAS  PubMed  Google Scholar 

  14. Diaz-Flores L, Gutierrez R, Madrid JF, Varela H, Valladares F, Acosta E, Martin-Vasallo P, Diaz-Flores L Jr (2009) Pericytes. Morphofunction, interactions and pathology in a quiescent and activated mesenchymal cell niche. Histol Histopathol 24:909–969

    CAS  PubMed  Google Scholar 

  15. Krueger M, Bechmann I (2010) CNS pericytes: concepts, misconceptions, and a way out. Glia 58:1–10

    Article  PubMed  Google Scholar 

  16. Guo M, Daines D, Tang J, Shen Q, Perrin RM, Takada Y, Yuan SY, Wu MH (2009) Fibrinogen-gamma C-terminal fragments induce endothelial barrier dysfunction and microvascular leak via integrin-mediated and RhoA-dependent mechanism. Arterioscler Thromb Vasc Biol 29:394–400

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Ward NL, Moore E, Noon K, Spassil N, Keenan E, Ivanco TL, LaManna JC (2007) Cerebral angiogenic factors, angiogenesis, and physiological response to chronic hypoxia differ among four commonly used mouse strains. J Appl Physiol 102:1927–1935

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by a grant from Heart and Stroke Foundation of Ontario (HSFO) and a scholarship (M.F.A.) from HSFO/Canadian Institute of Health Research (CIHR)/Canadian Stroke Network (CSN)/Astra Zeneca.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Freitas-Andrade, M., Slinn, J., Charlebois, C., Moreno, M.J. (2014). Histological Assessment of Angiogenesis in the Hypoxic Central Nervous System. In: Milner, R. (eds) Cerebral Angiogenesis. Methods in Molecular Biology, vol 1135. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0320-7_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0320-7_14

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0319-1

  • Online ISBN: 978-1-4939-0320-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics