Functional assay of mammalian and insect olfactory receptors using Xenopus oocytes.

The large number of olfactory receptors (ORs) expressed by various mammalian and insect species, as well as the large number of potential odorant ligands, has made the pairing of odorants with receptors -(de-orphaning) exceedingly difficult. These efforts are further complicated by difficulties in expressing ORs in many standard expression systems. Xenopus laevis oocytes offer a versatile expression platform for the de-orphaning and functional characterization of ORs. Two-electrode voltage clamp electrophysiology is a common and relatively straightforward approach to the functional assay of receptors expressed in Xenopus oocytes, and this technique has been discussed extensively in the literature. However, laboratories that are new to the use of Xenopus oocytes are often stymied by some of the peculiarities of the Xenopus oocyte expression system. We discuss some of the key methodological issues in Xenopus care, oocyte -isolation and receptor expression, with a focus on using this expression system to study the ORs of mammals and insects.