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Differentiation
1979 Jan 01;141-2:107-12. doi: 10.1111/j.1432-0436.1979.tb01018.x.
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Embryonic appearance of alpha, beta, and gamma crystallins in the periodic albinism (ap) mutant of Xenopus laevis.
McDevitt DS
,
Brahma SK
.
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The appearance of the crystallins during lens development in the periodic albinism (ap/ap) mutant of Xenopus laevis has been studied. Using antibodies specific for total crystallins, alpha + beta crystallins, and gamma crystallins in the immunofluorescence technique, the first positive reaction for all could be demonstrated in the Nieuwkoop-Faber Stage 31 lens rudiment. The antibody to alpha + beta crystallins exhibited differences in intensity from cell to cell in the early rudiment, while the reaction to the other antibodies was uniform throughout the rudiment. As lens differentiation progressed, immunofluorescence was restricted in all cases to the lens fiber area, up to and including Nieuwkas positive, however, for total lens crystallins. These results are at variance with earlier studies on lens development and the crystallins in wildtype (+/+) X. laevis, where a positive reaction for gamma and total crystallins could be detector total lens crystallins. That this divergence in the mutant is due to a pleiotropic effect or directly to the inductive failure of the endomesoderm to initiate melanogenesis, is discussed.
Fig. 1. Immunofluorescence [51 in 1.5% agar of: A and C adult X. Zaevis +/+ total lens proteins; B and D adult X. Zaevis aP/aP total lens proteins
against X. laevfs +/+ anti-total lens protein antibody (A and B) and X. Zaevis +/+ anti-cu + /4 crystallins antibody (C and D). Electrophoresis
performed at 4O C for 90 min, 250 V, 25 mA. Slides stained with Coomassie Brillant Blue after formation of the precipitin arcs. Arrow indicates
(I crystallin region, barely detectable in Xenopus due to its paucity in this organism [I61
Fig. 2. Photomicrographs of sections through the eye region of embryonic aP/aP X. luevis. Representative stages: A N-F Stage 31, as a histological
reference (hematoxylin and eosin). B-D, treated with anti-y crystallin antibody (darkfield immunofluorescence). B N-F Stage 3 1; C N-F
Stage 35/36; D N-F Stage 45. No immunofluorescence can be detected in the external layedlens epithelium. Identical immunofluorescence profdes
were obtained when N-F Stages 3 1-45 were incubated with anti-total lens protein antibody. Orientation: prospective retina to the left,
prospective cornea to the right. x 150
Fig. 3. Dark-field immunofluorescence photomicrograph of section
through the eye region of X. Zueuis aP/aP N-F Stage 31 embryo,
treated with anti-u + /3 crystallins antibody. Several cells in the rudiment
demonstrate intense positive immunofluorescence. Orientation:
prospective retina to the leR, prospective cornea to the right.
x 150
Rg. 4. Dark-field immunofluorescence photomicrograph of section
through the lens of one-year-old X. Zuevis aP/aP, treated with antitotal
lens protein antibody. The lens epithelium, the outermost single
layer of cells, exhibits positive immunofluorescence (nuclei of the lens
epithelial cells are negative). Orientation: retina to the left, cornea to
the right. x 43
