ABSTRACT
Ontogeny and localization of the lens crystallins, especially the γ-crystallins were investigated in Xenopus laevis lens regenerating system by the ‘indirect’ immunofluorescence staining method. Antibodies directed against Rana pipiens γ-crystallin antigen were used for the detection of this crystallin; the validity of such an experiment has been shown in a previous report. To detect total lens proteins we used X. laevis anti-total lens protein antibody.
The regenerates were staged according to Freeman (1963) and the first positive reaction with both the two antisera was observed in an early stage-4 regenerate. The site of the immunofluorescence reaction was nearly identical in both, suggesting that γ-crystallins are one of the first, if not the first of the lens crystallins to appear during lens regeneration.
The secondary fibres, when developed, showed less immunofluorescence than the primary fibres with R. pipiens anti-γ crystallin antibody, though the reaction was intense in the secondary fibres with X. laevis anti-total lens protein antibody.
The intensity and distribution of immunofluorescence increased with the growth of the lens. With the R. pipiens anti-γ crystallin antibody, the lens epithelium did not show any immunofluorescence reaction at any stage of lens regeneration. With X. laevis anti-total lens protein antibody, the epithelium showed an immunofluorescence reaction earlier than in the normal lens development. With the two antisera we used, we did not observe any immunofluorescence outside the lens tissue.