Three-dimensional structured illumination microscopy of liver sinusoidal endothelial cell fenestrations.
Three-dimensional structured illumination microscopy of liver sinusoidal endothelial cell fenestrations.
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DOI：
10.1016/j.jsb.2010.06.001
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发表时间：
2010-09
影响因子：
3
通讯作者：
Huser, Thomas R.
中科院分区：
文献类型：
--
作者：
Cogger, Victoria C.;McNerney, Gregory P.;Nyunt, Tun;DeLeve, Laurie D.;McCourt, Peter;Smedsrod, Bard;Le Couteur, David G.;Huser, Thomas R.
关键词：
开窗是肝窦内皮细胞中的孔，可过滤血液和肝细胞之间的基质和碎片。开窗在衰老和脂蛋白的调节中具有重要作用。然而，它们的小尺寸（<200 nm）禁止通过光学显微镜进行任何功能分析。我们采用结构照明光学显微镜以极高的清晰度和空间分辨率观察离体大鼠肝窦内皮细胞的开窗。利用该方法，阐明了开窗（直径123±24 nm）和筛板的三维结构，并表明开窗发生在细胞质突然变薄的区域（165±54 nm与非开窗区域的292±103 nm，P<0.0001）。筛板并不优先与荧光标记的F-肌动蛋白应力纤维和内皮一氧化氮合酶共定位，但似乎主要出现在细胞膜的衰减非筏区域中。未见迷宫式结构，所有开窗均为短圆柱形孔。总之，三维结构照明显微镜使荧光免疫染色和共定位的无限能力能够揭示有关开窗和筛板的新结构和功能信息。
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 colocalization to reveal new structural and functional information about fenestrations and sieve plates.
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DOI：
10.1152/ajpgi.00069.2008
发表时间：
2008-07-01
影响因子：
4.5
作者：
Cogger, Victoria C.;Arias, Irwin M.;Le Couteur, David G.
通讯作者：
Le Couteur, David G.
影响因子：
56.9
作者：
Schermelleh, Lothar;Carlton, Peter M.;Sedat, John W.
通讯作者：
Sedat, John W.
影响因子：
13.5
作者：
Warren, Alessandra;Le Couteur, David G.;Bertolino, Patrick
通讯作者：
Bertolino, Patrick
DOI：
10.1034/j.1600-0676.2001.021003198.x
发表时间：
2001-06-01
期刊：
LIVER
影响因子：
--
作者：
Yokomori, H;Oda, M;Ishii, H
通讯作者：
Ishii, H
DOI：
10.1111/j.1549-8719.2009.00004.x
发表时间：
2010-01
期刊：
Microcirculation (New York, N.Y. : 1994)
影响因子：
--
作者：
Warren A;Cogger VC;Arias IM;McCuskey RS;Le Couteur DG
通讯作者：
Le Couteur DG