Immunochemical characterization and quantitative distribution of crystallins in the epithelium and differentiating fibre cell populations of chick embryonic lens.

Lenses from 19-day chick embryos are fractionated by a double punch method to obtain the epithelium-annular pad complex (EP), outer fibres (OF), middle fibres (MF) and central fibres (CF). Water-soluble crystallins are characterized by SDS PAGE, isoelectric focusing (IEF) and two-dimensional IEF-SDS PAGE. Crystallins are also characterized by immunoelectrophoresis (IE), rocket IE, IEF-immunoblotting, and quantified by two-dimensional antigen-antibody crossed electrophoresis using antibodies to total 19-day embryonic as well as adult crystallins. In the adult lens, alpha-, beta- and delta-crystallins are 19%, 67% and 14%, respectively, while these are present at concentrations of 9%, 27% and 64%, respectively, in 19-day embryonic lens. In absolute amounts, delta-crystallin increases only by 1.23-fold between 19-day embryonic age and 6 months post-hatching, while total lens protein increases 12.5-fold. The predominance of delta-crystallin in central fibres, located along the optical axis, suggests that this protein is of embryonic origin. delta-Crystallin from fibres is electrofocused as 12 distinct molecular classes (pI 5.2-5.42) which react against anti-delta-crystallin on an immunoblot. Of these, the three most anodal species are not detected in EP. Fibres contain 50 000, 48 000 and 45 000 dalton delta-crystallin subunits while only 50 000 and 48 000 dalton subunits are present in EP.