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Updated: Aug 2, 2026
Assaying Surface Expression of Chemosensory Receptors in Heterologous Cells
Published on: February 23, 2011
Expression and intracellular localisation of odorant receptors in mammalian cell lines using Semliki Forest virus
K Monastyrskaia1, F Goepfert, R Hochstrasser
1Givaudan Roure Forschung AG, Dübendorf, Switzerland.
Abstract:
Odorant receptors are members of the G protein-coupled receptor superfamily. They are expressed on the surface of cilia of olfactory neurons, where they bind ligand (odorant). Studies of the molecular mechanisms of olfaction are complicated by the extremely large number of receptor genes, and difficulties in pairing a particular mammalian receptor to a specific odorant ligand in vivo. Here we report expression and localisation studies of two rat odorant receptor genes (17 and OR5), and C. elegans odr-10, using the Semliki Forest virus (SFV) system. All receptors were epitope-tagged at the N- or C-terminus in order to facilitate their detection in infected cells, and determine the localisation and membrane-orientation of recombinant proteins. The immortalised mouse olfactory neuronal cell line OLF 442, rat cortical and striatal primary neuron cultures, and the baby hamster kidney (BHK) cells, were infected and tested. Immunofluorescence and confocal microscopy studies performed on permeabilised, non-permeabilised and native cells revealed that in BHK cells the rat receptors 17 and OR5 were not targeted to the plasma membrane and remained in the endoplasmic reticulum. In contrast, in the mouse olfactory cell line OLF 442 both rat receptors were correctly inserted into the plasma membrane. Similar results were obtained using primary neurons, indicating that like mature neurons, the immortalised OLF 442 cells are capable of providing for correct odorant receptor processing and targeting.
Insights
Researchers studied odorant receptors (ORs) in different cell types. They found that specific mouse olfactory cells correctly process and target ORs, unlike other cell types, aiding olfaction research.
Area of Science:
- Molecular Biology
- Neuroscience
- Genetics
Background:
- Odorant receptors (ORs) are G protein-coupled receptors crucial for olfaction.
- Studying ORs is challenging due to the vast number of genes and difficulty in in vivo ligand pairing.
- Understanding OR expression and localization is key to elucidating olfactory mechanisms.
Purpose of the Study:
- To investigate the expression and localization of rat odorant receptors (17 and OR5) and C. elegans odr-10.
- To determine the correct membrane targeting and processing of these receptors in different cell systems.
- To assess the utility of the immortalized mouse olfactory cell line OLF 442 for OR studies.
Main Methods:
- Utilized the Semliki Forest virus (SFV) system for receptor gene expression.
- Epitope-tagged rat ORs (17, OR5) and C. elegans odr-10 for detection.
- Employed immunofluorescence and confocal microscopy in BHK cells, primary rat neuron cultures, and OLF 442 cells.
Main Results:
- Rat ORs 17 and OR5 were retained in the endoplasmic reticulum in baby hamster kidney (BHK) cells.
- In contrast, both rat ORs were correctly targeted to the plasma membrane in the OLF 442 mouse olfactory cell line.
- Primary rat neuron cultures also demonstrated correct receptor processing and targeting.
Conclusions:
- The immortalized OLF 442 cell line accurately recapitulates the plasma membrane targeting of odorant receptors observed in mature neurons.
- OLF 442 cells provide a reliable system for studying odorant receptor processing and localization.
- These findings facilitate research into the molecular mechanisms of olfaction.
