Confocal quantification of cis-regulatory reporter gene expression in living sea urchin.

Confocal quantification of cis-regulatory reporter gene expression in living sea urchin. Research Abstract Details 

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Confocal quantification of cis-regulatory reporter gene expression in living sea urchin. Abstract Text:

Sagar Damle,Bridget Hanser,Eric H Davidson,Scott E Fraser,

Quantification of GFP reporter gene expression at single cell level in living sea urchin embryos can now be accomplished by a new method of confocal laser scanning microscopy (CLSM). Eggs injected with a tissue-specific GFP reporter DNA construct were grown to gastrula stage and their fluorescence recorded as a series of contiguous Z-section slices that spanned the entire embryo. To measure the depth-dependent signal decay seen in the successive slices of an image stack, the eggs were coinjected with a freely diffusible internal fluorescent standard, rhodamine dextran. The measured rhodamine fluorescence was used to generate a computational correction for the depth-dependent loss of GFP fluorescence per slice. The intensity of GFP fluorescence was converted to the number of GFP molecules using a conversion constant derived from CLSM imaging of eggs injected with a measured quantity of GFP protein. The outcome is a validated method for accurately counting GFP molecules in given cells in reporter gene transfer experiments, as we demonstrate by use of an expression construct expressed exclusively in skeletogenic cells.

Confocal quantification of cis-regulatory reporter gene expression in living sea urchin. Publishing Authors By Initials

S Damle,B Hanser,EH Davidson,SE Fraser,

For similar animals: invertebrates: echinodermata: sea urchins research abstracts see: animals: invertebrates: echinodermata: sea urchins research

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Confocal quantification of cis-regulatory reporter gene expression in living sea urchin. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Developmental biology

VOLUME: 299

Page Numbers: 543-50

Journal Abbreviation: Dev. Biol.

ISSN: 0012-1606

DAY: 14

MONTH: 06

YEAR: 2006

Confocal quantification of cis-regulatory reporter gene expression in living sea urchin. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 372762

Confocal quantification of cis-regulatory reporter gene expression in living sea urchin. Keywords Mesh Terms:

KEYWORDS: Sea Urchins

MESH TERMS: metabolism

Chemical & Substance for Abstract: Confocal quantification of cis-regulatory reporter gene expression in living sea urchin. Information

Substance Name: Green Fluorescent Proteins

Registry Number: 147336-22-9

Grant and Affiliation Information for Confocal quantification of cis-regulatory reporter gene expression in living sea urchin.

AFFILIATION: Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA.

Country: United States

AGENCY: United States NICHD

GRANT: HD-37105

ACRONYM: HD

MEDLINETA: Dev Biol

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