| Using oocytes for Wnt signaling assays: paracrine assays and Wnt-conditioned medium. | |
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MedLine Citation:
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PMID: 20117213 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Xenopus oocytes have been widely used as a simple protein expression system particularly for the characterization of ion channels and membrane receptors. However, less attention has been given to their use as a means of synthesizing and analyzing secreted signaling molecules. In this review, we describe two assays that address this use of Xenopus oocytes. In the first, the paracrine assay, the oocytes secrete the signal and juxtaposed animal cap explants receive it. This provides an easy and efficient way to manipulate the signaling source since different doses of mRNA for the secreted ligand can be injected into the oocyte. Also the signaling response in the receiving cells can be read in several ways: in vivo by monitoring the localization of GFP-tagged signaling mediators, after fixation by immunostaining, or by monitoring changes in the transcriptional readout by RT-PCR. In a second approach, the oocyte is used to secrete a ligand, here the Wnt family members Wnt5a and 11, into the surrounding medium. This conditioned medium is then used to treat other cell lines to monitor their physiological changes in response to various combinations of Wnt proteins. Only a few recombinant Wnt proteins are commercially available and these are predominantly of mouse origin. Since Xenopus oocytes translate foreign RNA efficiently, this method provides an alternative source of Wnt protein derived from other model species. |
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Authors:
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Sang-wook Cha; Janet Heasman |
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Publication Detail:
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Type: Journal Article; Review Date: 2010-02-01 |
Journal Detail:
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Title: Methods (San Diego, Calif.) Volume: 51 ISSN: 1095-9130 ISO Abbreviation: Methods Publication Date: 2010 May |
Date Detail:
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Created Date: 2010-05-11 Completed Date: 2010-10-22 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9426302 Medline TA: Methods Country: United States |
Other Details:
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Languages: eng Pagination: 52-5 Citation Subset: IM |
Affiliation:
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Division of Developmental Biology, Department of Pediatrics, Cincinnati Children's Research Foundation, Cincinnati, OH 45229, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Algorithms Animals Cell Line Culture Media, Conditioned Female Green Fluorescent Proteins / metabolism Humans Ligands Mice Oocytes / metabolism* Recombinant Proteins / metabolism Reverse Transcriptase Polymerase Chain Reaction Signal Transduction Wnt Proteins / metabolism* Xenopus laevis |
| Chemical | |
Reg. No./Substance:
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0/Culture Media, Conditioned; 0/Ligands; 0/Recombinant Proteins; 0/Wnt Proteins; 147336-22-9/Green Fluorescent Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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