Three-dimensional structured illumination microscopy of liver sinusoidal endothelial cell fenestrations

Victoria C. Cogger, Gregory P. McNerney, Tun Nyunt, Laurie D. DeLeve, Peter McCourt, Bård Smedsrød, David G. Le Couteur, Thomas R Huser

Research output: Contribution to journalArticle

48 Scopus citations

Abstract

Fenestrations are pores in liver sinusoidal endothelial cells that filter substrates and debris between the blood and hepatocytes. Fenestrations have significant roles in aging and the regulation of lipoproteins. However their small size (<200. nm) has prohibited any functional analysis by light microscopy. We employed structured illumination light microscopy to observe fenestrations in isolated rat liver sinusoidal endothelial cells with great clarity and spatial resolution. With this method, the three-dimensional structure of fenestrations (diameter 123 ± 24. nm) and sieve plates was elucidated and it was shown that fenestrations occur in areas of abrupt cytoplasmic thinning (165 ± 54. nm vs. 292 ± 103. nm in non-fenestrated regions, P<0.0001). Sieve plates were not preferentially co-localized with fluorescently labeled F-actin stress fibers and endothelial nitric oxide synthase but appeared to occur in primarily attenuated non-raft regions of the cell membrane. Labyrinthine structures were not seen and all fenestrations were short cylindrical pores. In conclusion, three-dimensional structured illumination microscopy has enabled the unlimited power of fluorescent immunostaining and co-localization to reveal new structural and functional information about fenestrations and sieve plates.

Original languageEnglish (US)
Pages (from-to)382-388
Number of pages7
JournalJournal of Structural Biology
Volume171
Issue number3
DOIs
StatePublished - Sep 2010

Keywords

  • Actin
  • Fenestrations
  • Liver sinusoidal endothelial cell
  • Nitric oxide synthase
  • Structured illumination microscopy

ASJC Scopus subject areas

  • Structural Biology
  • Medicine(all)

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