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Fig. 1. Four-dimensional confocal analysis of Xl-ß-catenin-GFP expression.
(A-E) Frames from a time-lapse sequence following injection of
Xl-wt-ß-catenin-GFP mRNA at the 1-cell stage. Times after the start of
recording (hours:minutes) are shown in the bottom left corner of each panel
and cell number is shown in the bottom right corner. GFP-tagged ß-catenin
was initially localized in the nuclei, cytoplasm and junctional complexes of
all blastomeres (A). GFP-tagged ß-catenin disappeared from the animal
region of the embryo over a period of approximately two cell cycles (B-E).
GFP-tagged ß-catenin eventually became restricted to a small territory of
cells surrounding the vegetal pole (asterisk). (F-I) Frames from a time-lapse
sequence following injection of Xl-pt-ß-catenin-GFP at the 1-cell stage.
Mutation of residues phosphorylated by GSK3ß and a priming kinase at the
N-terminus of ß-catenin blocked the disappearance of GFP-tagged protein
from animal blastomeres. The vegetal pole is marked by an asterisk. (J)
Co-injection of mRNAs encoding Xl-wt-ß-catenin-GFP and a kinase-dead,
dominant negative form of GSK3ß (Xl-dnGSK3ß) at the 1-cell stage.
Expression of dnGSK3ß stabilized GFP-tagged ß-catenin in animal
blastomeres.
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