Ontogeny and localization of the a-, β-, and δ-crystallin antigens during lens development in mallard embryos

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Abstract
The ontogeny of α-, β-, and δ-crystallin antigens in the developing lens of mallard (Anaa platyrhynchos) was investigated by indirect immunofluorescence using specific antibodies to mallard α-, β-, and δ-crystallins raised in rabbits. The IgG fraction from each antiserum was isolated by affinity chromatography usign Protein A Sepharose CL-4.B. Fluoresceine (FITC) or rhodamine (TRITC) conjugated goat anti rabbit (GAR) γ-globulin was used as the secondary antibody. The results show that α-, β-, and δ-crystallins appear simultaneously in the developing mallard lens and are detectable from 66 hr (stage 15/16). This situation is different from the chick where δ-is known to appear first followed by β-, and α-. This could be due to species variation. In the epithelium, however, as in the chick, δ-emerges first followed by β-, and α-but rather late when compared with the chick and this seems probably due to a difference in the incubation time between the two species. Results from immunofluorescence and Tris-SDS gel electrophoresis demonstrate that αA subunit of the α-crystallin appears much earlier than the αB subunit. We also found that the newly discovered ε-crystallin (18) appears long after the three major crystallin classes.