Tracking Fenestrae Dynamics in Live Murine Liver Sinusoidal Endothelial Cells

The fenestrae of liver sinusoidal endothelial cells (LSECs) allow passive transport of solutes, macromolecules, and particulate material between the sinusoidal lumen and the liver parenchymal cells. Until recently, fenestrae and fenestrae‐associated structures were mainly investigated using electron...

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Published in:Hepatology (Baltimore, Md.) Vol. 69; no. 2; pp. 876 - 888
Main Authors: Zapotoczny, Bartlomiej, Szafranska, Karolina, Kus, Edyta, Braet, Filip, Wisse, Eddie, Chlopicki, Stefan, Szymonski, Marek
Format: Journal Article
Language:English
Published: United States Wolters Kluwer Health, Inc 01.02.2019
Subjects:
Animals
Atomic force microscopy
Cytochalasin B
Depsipeptides
Electron microscopy
Endothelial cells
Endothelial Cells - physiology
Functional morphology
Hepatocytes
Hepatology
Life span
Liver - cytology
Macromolecules
Mice
Microscopy
Microscopy, Atomic Force
Particulate matter
Solutes
ISSN:0270-9139, 1527-3350, 1527-3350
Online Access:Get full text
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Summary:The fenestrae of liver sinusoidal endothelial cells (LSECs) allow passive transport of solutes, macromolecules, and particulate material between the sinusoidal lumen and the liver parenchymal cells. Until recently, fenestrae and fenestrae‐associated structures were mainly investigated using electron microscopy on chemically fixed LSECs. Hence, the knowledge about their dynamic properties has remained to date largely elusive. Recent progress in atomic force microscopy (AFM) has allowed the study of live cells in three dimensions (X, Y, and Z) over a prolonged time (t) and this at unprecedented speeds and resolving power. Hence, we employed the latest advances in AFM imaging on living LSECs. As a result, we were able to monitor the position, size, and number of fenestrae and sieve plates using four‐dimensional AFM (X, Y, Z, and t) on intact LSECs in vitro. During these time‐lapse experiments, dynamic data were collected on the origin and morphofunctional properties of the filtration apparatus of LSECs. We present structural evidence on single laying and grouped fenestrae, thereby elucidating their dynamic nature from formation to disappearance. We also collected data on the life span of fenestrae. More especially, the formation and closing of entire sieve plates were observed, and how the continuous rearrangement of sieve plates affects the structure of fenestrae within them was recorded. We observed also the dawn and rise of fenestrae‐forming centers and defenestration centers in LSECs under different experimental conditions. Conclusion: Utilizing a multimodal biomedical high‐resolution imaging technique we collected fine structural information on the life span, formation, and disappearance of LSEC fenestrae; by doing so, we also gathered evidence on three different pathways implemented in the loss of fenestrae that result in defenestrated LSECs.
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ISSN:0270-9139
1527-3350
1527-3350
DOI:10.1002/hep.30232