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Development, Vol 124, Issue 22 4649-4659, Copyright © 1997 by Company of Biologists
JOURNAL ARTICLES |
MI Arnone, LD Bogarad, A Collazo, CV Kirchhamer, RA Cameron, JP Rast, A Gregorians and EH Davidson
Division of Biology and Stowers Institute for Medical Research, California Institute of Technology, Pasadena 91125, USA.
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.
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L. A. Romano and G. A. Wray Conservation of Endo16 expression in sea urchins despite evolutionary divergence in both cis and trans-acting components of transcriptional regulation Development, September 1, 2003; 130(17): 4187 - 4199. [Abstract] [Full Text] [PDF]
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E. Davidson, R. Cameron, and A Ransick Specification of cell fate in the sea urchin embryo: summary and some proposed mechanisms Development, January 9, 1998; 125(17): 3269 - 3290. [Abstract] [PDF]
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M. Arnone, E. Martin, and E. Davidson Cis-regulation downstream of cell type specification: a single compact element controls the complex expression of the CyIIa gene in sea urchin embryos Development, January 4, 1998; 125(8): 1381 - 1395. [Abstract] [PDF]
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