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Dev Biol
1996 Jan 10;1731:348-52. doi: 10.1006/dbio.1996.0029.
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Analysis of Wnt/Engrailed signaling in Xenopus embryos using biolistics.
Koster JG
,
Eizema K
,
Peterson-Maduro LJ
,
Stegeman BI
,
Destrée OH
.
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We adapted the Biolistics Particle Delivery System for the introduction of DNA into Xenopus embryos, allowing us to modulate the expression of different genes at specific time points during development. In the present study we applied the Biolistics method to the study of the wnt-engrailed signaling cascade in the developing Xenopus embryo. We show that ectopic expression of Xwnt-1 and Xwnt-5C is sufficient to activate specifically XEn-1 and not XEn-2.
FIG. 1. Expression of DNA constructs after introduction into Xenopus laevis embryos by the Biolistics technique and effects of ectopic
Xwnt expression on XEn-1 and XEn-2 expression. DNA constructs were bombarded into neurula stage embryos (stage 19), analysis was
at late tailbud stage (stage 33). Anterior is to the right, dorsal is upward. (A–D) Views of Xwnt-5C-bombarded stage-33 embryos probed
with XEn-1 (A and C), XEn-2 (B), and Xwnt-5C RNA (D), respectively. (C) A higher magnification of the trunk of the embryo shown in
A to evidence gold particles (open arrowheads) in an area of ectopic expression. Endogenous XEn-1 and XEn-2 hybridization signals are
detected (A and B): At the mid/hindbrain border (XEn-1 and XEn-2), in a group of ventrolateral cells in the spinal cord (XEn-1), in the
anterior part of the pronephros (XEn-1), and in the branchial arches (XEn-2) (Eizema et al., 1994). The endogenous Xwnt-5C RNA is
detectable in the tail and the mediodorsal part of the mid/hindbrain region of the central nervous system (D) (unpublished results). (E)
Embryos bombarded with a b-galactosidase reporter gene construct at neurula stage and assayed for b-galactosidase activity at late tailbud
stage.
