Nuclear microinjection to assess how heterologously expressed proteins impact Ca2+ signals in Xenopus oocytes. Cold Spring Harb Protoc 2013 Mar 01;2013(3)
Date
03/05/2013Pubmed ID
23457340Pubmed Central ID
PMC4021487DOI
10.1101/pdb.prot072785Scopus ID
2-s2.0-84874687582 (requires institutional sign-in at Scopus site) 5 CitationsAbstract
The Xenopus oocyte is frequently used for heterologous expression and for studying the spatiotemporal patterning of Ca(2+) signals. Here, we outline a protocol for nuclear microinjection of the Xenopus oocyte for the purpose of studying how subsequently expressed proteins impact intracellular Ca(2+) signals evoked by inositol trisphosphate (InsP3). Injected oocytes can easily be identified by reporter technologies and the impact of heterologously expressed proteins on the generation and properties of InsP3-evoked Ca(2+) signals can be resolved using caged InsP3 and fluorescent Ca(2+) indicators.
Author List
Lin-Moshier Y, Marchant JSAuthor
Jonathan S. Marchant PhD Chair, Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsCalcium
Inositol Phosphates
Microinjections
Molecular Biology
Oocytes
Recombinant Proteins
Signal Transduction
Xenopus