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Windoffer, R., Beile,
B., Leibold, A., Thomas, S., Wilhelm, U., and Leube, R.E. 2000.
Visualization of gap junction mobility in living cells.
Cell Tissue Res. 299:347-362. | abstract | pdf | | 12 |
download.mov (5 MB)
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movie 1. Time-lapse
fluorescence microscopy documenting dynamics of Cx.EGFP-1-positive
structures in living PLC cells of clone PCx-9. Note the different types
of mobility of fluorescent patches at cell contact sites. A region of
high mobility is labeled by rectangle, fusion of plaques is demarcated
by ovoid between 0.00 min and 9.50 min and again in another area by
ovoid between 9.75 min and 28 min. Separation of a plaque into several
fragments is marked by circle. The time points of recording are given
at lower left corner in min. Some strongly fluorescent, large
structures that remain stationary are seen in the cytoplasm. Note also
the presence of many rapidly moving, small cytoplasmic dots.
Pictures taken every 15 seconds for 28 minutes. |
download.mov (4,2 MB)
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movie 2. Fluorescence
micrographs (inverse presentation) depicting dynamic heterogeneities in
two Cx.EGFP-1-positive cell contact sites that are viewed en face in
live PLC cells of clone PCx-9. The time of recording is given in the
upper left corner. Note the high degree of mobility of patches with
differing fluorescence intensity within the gap junctions shown. In
addition, weak and highly mobile fluorescence is also detectable in the
vicinity corresponding to cytoplasmic regions.
Pictures taken every 8.4 seconds for 9.94 minutes. |
download.mov (0,7 MB)
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movie 3. Time-lapse
fluorescence microscopy of Cx.EGFP-1-expressing PCx-9 cells depicting
endocytosis of gap junctional fragments. The sequence shows budding of
a tubular structure from an extensive junction (arrow).
Pictures taken every 60 seconds for 24 minutes. |
download.mov (1,8 MB)
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movie 4. Time-lapse
fluorescence microscopy of Cx.EGFP-1-expressing PCx-9 cells depicting
endocytosis of gap junctional fragments. A strongly fluorescent patch
(arrow) can be tracked within a gap junction which first extends into
the cytoplasm (9.5 min), retracts (20.5 min) before it eventually buds
off (22.5 min) and is taken up into the cytoplasm (27 min).
Pictures taken every 30 seconds for 27 minutes. |
download.mov (21 MB)
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movie 5. Series
of overlay pictures showing brightfield pictures merged with
fluorescence pictures (green color) to demonstrate Cx.EGFP-1 mobility
in four PCx-9 cells before and after addition of cycloheximide (17.1
µM). The cells remained viable during the entire period of 249 min as
judged from the ongoing undulations of peripheral cell regions and the
continuous fast movements of cytoplasmic dots. Note the stationary
nature of large cytoplasmic vesicular structures contrasting with the
highly dynamic plaques at cell contact sites which are subject to
continuous remodeling. Between 199 min and 204 min, i.e., almost 3 h
after addition of cycloheximide a large plaque is broken up (upper
left). Concurrently, a cell slides between the two cells establishing
new contacts. Note the overall reduction in fluorescent structures
during cycloheximide treatment, most notably at cell contact sites. Pictures taken every 60 seconds for 249 minutes. |
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