Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 187-202 |
| Number of pages | 16 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 1003 |
| DOIs | |
| State | Published - Oct 4 2013 |
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ASJC Scopus subject areas
- Medicine(all)
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Functional assay of mammalian and insect olfactory receptors using Xenopus oocytes. / Luetje, Charles W; Nichols, Andrew S.; Castro, Ana; Sherman, Benjamin L.
In: Methods in molecular biology (Clifton, N.J.), Vol. 1003, 04.10.2013, p. 187-202.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Functional assay of mammalian and insect olfactory receptors using Xenopus oocytes.
AU - Luetje, Charles W
AU - Nichols, Andrew S.
AU - Castro, Ana
AU - Sherman, Benjamin L.
PY - 2013/10/4
Y1 - 2013/10/4
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=84884744407&partnerID=8YFLogxK
U2 - 10.1007/978-1-62703-377-0_14
DO - 10.1007/978-1-62703-377-0_14
M3 - Article
C2 - 23585043
AN - SCOPUS:84884744407
VL - 1003
SP - 187
EP - 202
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
SN - 1064-3745
ER -