We have used a combination of immunogold staining, optical sectioning light microscopy, intermediate voltage electron microscopy, and EM tomography to examine the distribution of lamin B over the nuclear envelope of CHO cells. Apparent inconsistencies between previously published light and electron microscopy studies of nuclear lamin staining were resolved. At light microscopy resolution, an apparent open fibrillar network is visualized. Colocalization of lamin B and nuclear pores demonstrates that these apparent fibrils, separated by roughly 0.5 micron, are anti-correlated with the surface distribution of nuclear pores; pore clusters lie between or adjacent to regions of heavy lamin B staining. Examination at higher, EM resolution reveals that this apparent lamin B network does not correspond to an actual network of widely spaced, discrete bundles of lamin filaments. Rather it reflects a quantitative variation in lamin staining over a roughly 0.5-micron size scale, superimposed on a more continuous but still complex distribution of lamin filaments, spatially heterogeneous on a 0.1-0.2-micron size scale. Interestingly, lamin B staining at this higher resolution is highly correlated to the underlying chromatin distribution. Heavy concentrations of lamin B directly "cap" the surface of envelope associated, large-scale chromatin domains.
Article|
December 15 1993
Lamin B distribution and association with peripheral chromatin revealed by optical sectioning and electron microscopy tomography.
A S Belmont
,
A S Belmont
Department of Cell and Structural Biology, University of Illinois, Champaign-Urbana 61801.
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Y Zhai
,
Y Zhai
Department of Cell and Structural Biology, University of Illinois, Champaign-Urbana 61801.
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A Thilenius
A Thilenius
Department of Cell and Structural Biology, University of Illinois, Champaign-Urbana 61801.
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A S Belmont
Department of Cell and Structural Biology, University of Illinois, Champaign-Urbana 61801.
Y Zhai
Department of Cell and Structural Biology, University of Illinois, Champaign-Urbana 61801.
A Thilenius
Department of Cell and Structural Biology, University of Illinois, Champaign-Urbana 61801.
Online Issn: 1540-8140
Print Issn: 0021-9525
J Cell Biol (1993) 123 (6): 1671–1685.
Citation
A S Belmont, Y Zhai, A Thilenius; Lamin B distribution and association with peripheral chromatin revealed by optical sectioning and electron microscopy tomography.. J Cell Biol 15 December 1993; 123 (6): 1671–1685. doi: https://doi.org/10.1083/jcb.123.6.1671
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