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Fig. 2. Identification of the regulatory elements required for telencephalic
expression of the –8.4ngn1:gfp transgene in the zebrafish.
(A,B) Analysis of zebrafish embryos transgenic for truncated versions of the
–8.4ngn1:gfp reporter fragment. Transgene expression in the
telencephalon (arrows) is no longer detected in embryos carrying transgenes
with 5.3 kb of ngn1 sequence. (C) Overlapping deletions spanning the
region between 6.9 and 5.3 kb reveal a 200-bp element, the LSE, that
is required for telencephalic expression of the –8.4 ngn1:gfp
transgene. The heads of embryos in A are seen from the dorsal aspect, with the
telencephalon up, and are staged between 28 and 30 hpf. Boxes in B,
corresponding to the LSE (blue) and LATE (green), indicate
regions of homology between the zebrafish, mouse and human ngn1 loci.
Each construct was analysed in one to three transgenic lines. Note that
expression patterns from the overlapping deletions provided an independent
further confirmation of the results. Transgenic lines varied in the intensity
of reporter expression, but no significant variations in the expression
pattern due to integration site effects were observed.
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