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Evaluating cell-surface expression and measuring activation of mammalian odorant receptors in heterologous cells.

Publication ,  Journal Article
Zhuang, H; Matsunami, H
Published in: Nat Protoc
2008

A fundamental question in olfaction is which odorant receptors (ORs) are activated by a given odorant. A major roadblock to investigating odorant-OR relationships in mammals has been the inability to express ORs in heterologous cells suitable for screening active ligands for ORs. The discovery of the receptor-transporting protein family has facilitated the effective cell-surface expression of ORs in heterologous cells. The establishment of a robust heterologous expression system for mammalian ORs facilitates the high-throughput 'deorphanization' of these receptors by matching them to their cognate ligands. This protocol details the method used for evaluating the cell-surface expression and measuring the functional activation of ORs of transiently expressed mammalian ORs in HEK293T cells. The stages of OR cell-surface expression include cell culture preparation, transfer of cells, transfection, immunocytochemistry or flow cytometry, odorant stimulation and luciferase assay. This protocol can be completed in a period of 3 d from the transfer of cells to cell-surface expression detection and/or measurement of functional activation.

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Published In

Nat Protoc

DOI

EISSN

1750-2799

Publication Date

2008

Volume

3

Issue

9

Start / End Page

1402 / 1413

Location

England

Related Subject Headings

  • Smell
  • Signal Transduction
  • Receptors, Odorant
  • Molecular Biology
  • Luciferases
  • Ligands
  • Immunohistochemistry
  • Humans
  • Flow Cytometry
  • Cell Line
 

Citation

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Zhuang, H., & Matsunami, H. (2008). Evaluating cell-surface expression and measuring activation of mammalian odorant receptors in heterologous cells. Nat Protoc, 3(9), 1402–1413. https://doi.org/10.1038/nprot.2008.120
Zhuang, Hanyi, and Hiroaki Matsunami. “Evaluating cell-surface expression and measuring activation of mammalian odorant receptors in heterologous cells.Nat Protoc 3, no. 9 (2008): 1402–13. https://doi.org/10.1038/nprot.2008.120.
Zhuang, Hanyi, and Hiroaki Matsunami. “Evaluating cell-surface expression and measuring activation of mammalian odorant receptors in heterologous cells.Nat Protoc, vol. 3, no. 9, 2008, pp. 1402–13. Pubmed, doi:10.1038/nprot.2008.120.

Published In

Nat Protoc

DOI

EISSN

1750-2799

Publication Date

2008

Volume

3

Issue

9

Start / End Page

1402 / 1413

Location

England

Related Subject Headings

  • Smell
  • Signal Transduction
  • Receptors, Odorant
  • Molecular Biology
  • Luciferases
  • Ligands
  • Immunohistochemistry
  • Humans
  • Flow Cytometry
  • Cell Line