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Published June 1992 | public
Journal Article

Territorial Expression of Three Different trans-Genes in Early Sea Urchin Embryos Detected by a Whole-Mount Fluorescence Procedure


We have developed a new procedure for detection of the protein product of chloramphenicol acetyltransferase (CAT) reporter genes in whole mounted sea urchin embryos. The position of a commercially available anti-CAT antibody is visualized by video or confocal microscopy, and thus the spatial domains of exogenous reporter gene expression can be determined with regard to the intact three-dimensional structures of the embryo. We show that in pluteus stage embryos CAT protein expression patterns for SM50·CAT or CyIIIɑ·CAT reporter genes are similar to those previously obtained by in situ hybridizations with radioactive probes. Taking advantage of the superior resolution of cellular CAT expression patterns using the antibody visualization method, we found for the first time that, in addition to the expression in aboral ectoderm, some cells in the ciliated band of the pluteus express CyIIIɑ·CAT. The expression of a new fusion construct, CyIIɑ·CAT, was also examined. As expected from the localization of endogenous CyIIɑ mRNA, CAT protein was expressed under control of the CyIIɑ promoter in gut and skeletogenic mesenchyme cells.

Additional Information

© 1992 Academic Press, Inc. Accepted February 24, 1992. We thank Dr. Scott E. Fraser for the use of the confocal microscope. This research was supported by NIH Grant HD-05753 to E.H.D. and NSF Grant DCB-8911829 to R.A.C. R.W.Z. is supported by NIH Training Grant GM-07616.

Additional details

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October 19, 2023