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Fig. 1. The upstream (5402-1473 bp) and the first intron (812-24 bp) regions are
capable of directing Hr-Otx transcription during cleavage stages. (A)
The structure of 5' region of Hr-Otx, including the putative
endogenous promoter (TATAA) and intron 1. Boxes on the gray line indicate
exons; ATG indicates the translation start site, which is located in the
second exon. The light gray lines are the genomic regions examined for their
transcription-driving activity. (B,B',D,D') Expression of
Hr-Otx detected by whole-mount in situ hybridization.
(C,C',E-F') Transcription of lacZ in embryos injected
with the reporter construct harboring the upstream region 5402-1473
(C,C',E,E'), or the intron region 812-24 (F,F'), as
visualized by in situ hybridization. In injected embryos, not all
Hr-Otx-expressing cells are stained because of the mosaic inheritance
of injected DNA. Specimens at the 32-cell stage (B-C') and 64-cell stage
(D-F') are shown. (B-F) Animal view; (B'-F') vegetal view. A
schematic representation of in situ hybridization is shown on the right side
of each specimen (C,C',E-F'). Cells expressing Hr-Otx are
colored, according to their derivation from the 8-cell embryo (H), as follows:
a-line, dark green; b-line, light green; A-line, orange; and B-line, yellow.
Blue dots indicate the cells in which lacZ transcripts were detected.
Below the specimens of whole-mount in situ hybridization, the identity of the
lacZ-transcribing cell line is indicated. (G) Frequency of
lacZ -positive cells for a given cell line in embryos injected with
the construct harboring the region 5402-1473bp or 812-23bp, examined at the
32-cell stage (top) and 64-cell stage (bottom). Color codes are as in H. Gray
indicates ectopic expression. Number in parenthesis indicates number of
embryos examined.
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