Abstract
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, α-, β- and δ-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, δ-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 δ-crystallin in central fibres, located along the optical axis, suggests that this protein is of embryonic origin.δ-Crystallin from fibres is electrofocused as 12 distinct molecular classes (pI 5·2–5·42) which react against anti-δ-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 δ-crystallin subunits while only 50 000 and 48 000 dalton subunits are present in EP.
Published Version
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