Arnone, Maria I. and Bogarad, Leonard D. and Collazo, Andres and Kirchhamer, Carmen V. and Cameron, R. Andrew and Rast, Jonathan P. and Gregorians, Armenia and Davidson, Eric H. (1997) Green Fluorescent Protein in the sea urchin: new experimental approaches to transcriptional regulatory analysis in embryos and larvae. Development, 124 (22). pp. 4649-4659. ISSN 0950-1991. http://resolver.caltech.edu/CaltechAUTHORS:20120118-112101095
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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.
|Additional Information:||© 1997 The Company of Biologists Limited. Accepted 5 September 1997. This work was supported by the Stowers Institute for Medical Research, and by grants to E. H. D. from the National Institutes of Child Health and Human Development, and the Division of Research Resources of the NIH. J. P. R. is supported by an NRSA Fellowship. L. D. B. is supported by a Gosney Fellowship.|
|Subject Keywords:||gene regulation; transgenic analysis; larval development; mosaic DNA incorporation; Green Fluorescent Protein (GFP); sea urchin|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Tony Diaz|
|Deposited On:||19 Jan 2012 19:31|
|Last Modified:||26 Dec 2012 14:42|
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