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 Ca2+ signals. Here, we outline a protocol for nuclear microinjection of the Xenopus oocyte for the purpose of studying how subsequently expressed proteins impact intracellular Ca2+ 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 Ca2+ signals can be resolved using caged InsP3 and fluorescent Ca2+ indicators.