Membrane skeletons in avian erythrocytes as revealed by the quick-freezing and deep-etching method

Ultrastructure of chicken erythrocytes were examined by the quick-freezing and deep-etching (QF-DE) method. Some erythrocytes were fixed with paraformaldehyde and prepared with erythrocyte-splitting method or saponin treatment to remove soluble proteins before quick-freezing. Others were prepared in the cytosol buffer with the erythrocyte-splitting method to obtain natural state of cytoskeletons. Non-expanding membrane skeletons were highly condensed on the cytoplasmic side of lipid membrane in the paraformaldehyde-fixed specimens. Under unilateral extension of the specimens, long stretched filaments were connected alternately with condensed filamentous or granular structures under erythrocyte membranes. As the membrane skeletons got closer to the marginal bands, they become more dense network structures. Moreover, in the fresh unfixed specimens, dense networks of filaments were localized underlying erythrocyte membranes in a relatively intact state. Fine filaments connected the marginal microtubule bands to the cytoplasmic sides of erythrocyte membranes. The different distribution of each cytoskeletal component and the association of these structures may support the elliptocytic shape of chicken erythrocytes and resist the dynamic circumstance.