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David C. Beebe, Oleg Vasiliev, Jianli Guo, Ying-Bo Shui, Steven Bassnett; Changes in Adhesion Complexes Define Stages in the Differentiation of Lens Fiber Cells. Invest. Ophthalmol. Vis. Sci. 2001;42(3):727-734.
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purpose. During their differentiation, lens fiber cells elongate, detach from the lens
capsule, associate at the sutures, and degrade all cytoplasmic
membrane-bound organelles. Changes in the expression or organization of
cell adhesion and cytoskeleton-associated proteins were correlated
with these events during fiber cell differentiation in chicken embryos.
methods. Fixed or living lenses were sliced with a tissue slicer, permeabilized
or extracted with detergents, stained with antibodies or
fluorescent-labeled phalloidin, and viewed with a confocal microscope.
The distribution of N-cadherin in elongating and mature fiber cells was
determined by Western blot analysis. Reverse transcription–polymerase
chain reaction (RT-PCR) was used to determine the distribution of
vinculin and paxillin transcripts.
results. Staining for N-cadherin and band 4.1 protein decreased soon after fiber
cells detached from the capsule. Detergent extraction of lens sections
and Western blots of dissected lens regions showed that much of this
decrease in staining was due to epitope masking. Vinculin
immunoreactivity was barely detectable on the lateral membranes of
elongating fiber cells but increased markedly once these cells reached
their maximum length and formed the sutures. Staining for paxillin was
also low in elongating fiber cells but increased late in fiber cell
differentiation, just before the cells destroyed their membrane-bound
organelles. Spectrin and ankyrin immunoreactivity did not change when
fiber cells reached the sutures. Staining for F-actin increased
transiently in cells that had just reached the sutures. Messenger RNAs
for vinculin and paxillin were more abundant in maturing than in
elongating fiber cells.
conclusions. The adhesion complexes of lens fiber cells change in organization and
composition soon after these cells finish elongating and detach from
the capsule. Increased staining for vinculin and paxillin defines
distinct stages of fiber cell differentiation that are intermediate
between the completion of cell elongation and the time when lens fiber
cells degrade their membrane-bound organelles. Remodeling adhesion
complexes during fiber cell maturation may assure the stability of
fiber–fiber associations, once these cells are no longer
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