A quick-freezing and deep-etching method in combination with erythrocyte
splitting was used to examine
the cytoplasmic aspect of whole-mount human erythrocyte membranes. Various
external forces induced
alterations in membrane skeletal organisation during the splitting procedure.
The initial change was
elongation in the peripheral part of the membrane skeleton, examined by
immunostaining with a
monoclonal antispectrin antibody. Under severe stretching conditions, a
linear
rearrangement of filamentous
components was evident; these were disposed parallel to the rim of the
erythrocyte, while the central part of
the concavity exhibited a more compacted structure. These changes resulted
in a
different distribution of
membrane skeletal components between central rigid and peripheral flexible
areas
in biconcave erythrocytes.
It is suggested that the reversible membrane skeletal changes in the flexible
areas which resist the external
forces are important for maintaining the normal framework of biconcave
human
erythrocytes.