Preparation and characterization of native lens cell cytoplasm

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Abstract
We have developed an ultracentrifugation procedure for isolating fractions of native cortical and nuclear cytoplasm from calf lens without dilution and free of cell membranes. The concentration of these isolated cytoplasmic fractions spans the physiological range (20 to 50%). Some fractions of the nuclear cytoplasm phase-separate at low temperature giving rise to opacification while other fractions do not phase-separate. The fractions of cortical cytoplasm do not phase-separate at any temperature. Both cortical and nuclear cytoplasm were studied using quasielastic light scattering spectroscopy, electron microscopy, SDS-polyacrylamide gel electrophoresis and isoelectric focusing. We found that the structural elements in the cortical cytoplasm were very similar to those in the nuclear cytoplasm.