The use of Green Fluorescent Protein (GFP) as a reporter
for expression transgenes opens the way to several new
experimental strategies for the study of gene regulation in
sea urchin development. A GFP coding sequence was associated
with three different previously studied cis-regulatory
systems, viz those of the SM50 gene, expressed in skeletogenic mesenchyme, the CyIIa gene, expressed in archenteron, skeletogenic and secondary mesenchyme, and the
Endo16 gene, expressed in vegetal plate, archenteron and
midgut. We demonstrate that the sensitivity with which
expression can be detected is equal to or greater than that
of whole-mount in situ hybridization applied to detection
of CAT mRNA synthesized under the control of the same
cis-regulatory systems. However, in addition to the
important feature that it can be visualized nondestructively
in living embryos, GFP has other advantages. First, it freely diffuses even within fine cytoplasmic cables, and thus reveals connections between cells, which in sea urchin
embryos is particularly useful for observations on regulatory systems that operate in the syncytial skeletogenic mesenchyme. Second, GFP expression can be dramatically visualized in postembryonic larval tissues. This brings postembryonic larval developmental processes for the first time within the easy range of gene transfer analyses. Third, GFP permits identification and segregation of embryos in which the clonal incorporation of injected DNA has occurred in any particular desired region of the embryo. Thus, we show explicitly that, as expected, GFP transgenes are incorporated in the same nuclei together with other transgenes with which they are co-injected
