title: Nuclear microinjection to assess how heterologously expressed proteins impact Ca2+ signals in Xenopus oocytes.
abstract: 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.
authors: Department of Pharmacology, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
authors: Yaping
authors: Y
authors: Lin-Moshier
authors: Jonathan S
authors: JS
authors: Marchant
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D007295:Inositol Phosphates / Q000378:metabolism; D008845:Microinjections / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D009865:Oocytes / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D015398:Signal Transduction; D014981:Xenopus
chemical_list: D007295:Inositol Phosphates; D011994:Recombinant Proteins; C516441:inositol trispyrophosphate; D002118:Calcium