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High-Resolution Confocal Imaging of Pericytes in Human Fetal Brain Microvessels

  • Mariella ErredeEmail author
  • Francesco Girolamo
  • Daniela Virgintino
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Part of the Methods in Molecular Biology book series (MIMB, volume 2206)

Abstract

体育赛事投注记录pericytes are integral part of neurovascular unit and play a role in the maintenance of blood-brain barrier integrity, angiogenesis, and cerebral blood flow regulation. despite their important functional roles, a univocal phenotypic identification is still emerging also for the lack of a “pan-pericyte” marker. in the present study, we describe in detail the method for performing fluorescence immunohistochemistry on thick free-floating sections from human fetal brain in high resolution laser confocal microscopy. this method enables to obtain three-dimensional images of pericytes and provides insights about their distribution and localization in the microvessels of human developing brain.

Key words

Pericyte Vibratome sectioning High-resolution confocal microscopy 

Notes

Acknowledgments

the authors gratefully acknowledge prof. roberto perris for the generous ng 22161d7 antibody gift.

References

  1. 1.
    Armulik A, Abramsson A, Betsholtz C (2005) Endothelial/pericyte interactions. Circ Res 97(6):512–523
  2. 2.
    Attwell D et al (2016) What is a pericyte? J Cereb Blood Flow Metab 36(2):451–455
  3. 3.
    Sweeney MD, Ayyadurai S, Zlokovic BV (2016) Pericytes of the neurovascular unit: key functions and signaling pathways. Nat Neurosci 19(6):771–783
  4. 4.
    McConnell HL et al (2017) The translational significance of the neurovascular unit. J Biol Chem 292(3):762–770
  5. 5.
    Virgintino D et al (2007) An intimate interplay between precocious, migrating pericytes and endothelial cells governs human fetal brain angiogenesis. Angiogenesis 10(1):35–45
  6. 6.
    Ribatti D, Nico B, Crivellato E (2011) The role of pericytes in angiogenesis. Int J Dev Biol 55(3):261–268
  7. 7.
    Chiaverina G et al (2019) Dynamic interplay between pericytes and endothelial cells during sprouting angiogenesis. Cells 8(9)
  8. 8.
    Armulik A et al (2010) Pericytes regulate the blood-brain barrier. Nature 468(7323):557–561
  9. 9.
    Berthiaume AA et al (2018) Pericyte structural remodeling in cerebrovascular health and homeostasis. Front Aging Neurosci 10:210
  10. 10.
    Dalkara T, Gursoy-Ozdemir Y, Yemisci M (2011) Brain microvascular pericytes in health and disease. Acta Neuropathol 122(1):1–9
  11. 11.
    Jansson D et al (2014) A role for human brain pericytes in neuroinflammation. J Neuroinflammation 11:104
  12. 12.
    Armulik A, Genove G, Betsholtz C (2011) Pericytes: developmental, physiological, and pathological perspectives, problems, and promises. Dev Cell 21(2):193–215
  13. 13.
    Blinder P et al (2013) The cortical angiome: an interconnected vascular network with noncolumnar patterns of blood flow. Nat Neurosci 16(7):889–897
  14. 14.
    Zlokovic BV (2008) The blood-brain barrier in health and chronic neurodegenerative disorders. Neuron 57(2):178–201
  15. 15.
    Hartmann DA et al (2015) Pericyte structure and distribution in the cerebral cortex revealed by high-resolution imaging of transgenic mice. Neurophotonics 2(4):041402
  16. 16.
    Bonkowski D et al (2011) The CNS microvascular pericyte: pericyte-astrocyte crosstalk in the regulation of tissue survival. Fluids Barriers CNS 8(1):8
  17. 17.
    Dore-Duffy P, Cleary K (2011) Morphology and properties of pericytes. Methods Mol Biol 686:49–68
  18. 18.
    Smyth LCD et al (2018) Markers for human brain pericytes and smooth muscle cells. J Chem Neuroanat 92:48–60
  19. 19.
    Krueger M, Bechmann I (2010) CNS pericytes: concepts, misconceptions, and a way out. Glia 58(1):1–10
  20. 20.
    Winkler EA, Bell RD, Zlokovic BV (2010) Pericyte-specific expression of PDGF beta receptor in mouse models with normal and deficient PDGF beta receptor signaling. Mol Neurodegener 5:32
  21. 21.
    Nehls V, Drenckhahn D (1991) Heterogeneity of microvascular pericytes for smooth muscle type alpha-actin. J Cell Biol 113(1):147–154
  22. 22.
    Bandopadhyay R et al (2001) Contractile proteins in pericytes at the blood-brain and blood-retinal barriers. J Neurocytol 30(1):35–44
  23. 23.
    Chen J et al (2017) CD146 coordinates brain endothelial cell-pericyte communication for blood-brain barrier development. Proc Natl Acad Sci U S A 114(36):E7622–E7631
  24. 24.
    Alliot F et al (1999) Pericytes and periendothelial cells of brain parenchyma vessels co-express aminopeptidase N, aminopeptidase A, and nestin. J Neurosci Res 58(3):367–378
  25. 25.
    Ozerdem U et al (2001) NG2 proteoglycan is expressed exclusively by mural cells during vascular morphogenesis. Dev Dyn 222(2):218–227
  26. 26.
    Stallcup WB (2002) The NG2 proteoglycan: past insights and future prospects. J Neurocytol 31(6-7):423–435
  27. 27.
    Tamburini E et al (2019) Structural deciphering of the NG2/CSPG4 proteoglycan multifunctionality. FASEB J 33(3):3112–3128
  28. 28.
    Girolamo F et al (2013) Diversified expression of NG2/CSPG4 isoforms in glioblastoma and human foetal brain identifies pericyte subsets. PLoS One 8(12):e84883
  29. 29.
    Winkler EA, Bell RD, Zlokovic BV (2011) Central nervous system pericytes in health and disease. Nat Neurosci 14(11):1398–1405
  30. 30.
    Biggs DS (2010) 3D deconvolution microscopy. Curr Protoc Cytom Chapter 12:Unit 12.19 1–Unit 12.1920

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Mariella Errede
    • 1
    Email author
  • Francesco Girolamo
    • 1
  • Daniela Virgintino
    • 1
  1. 1.Human Anatomy and Histology Unit, Department of Basic Medical Sciences, Neurosciences and Sensory OrgansBari University School of MedicineBariItaly

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