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Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase.

Publication ,  Journal Article
de March, CA; Fukutani, Y; Vihani, A; Kida, H; Matsunami, H
Published in: J Vis Exp
April 23, 2019

Olfactory perception begins with the interaction of odorants with odorant receptors (OR) expressed by olfactory sensory neurons (OSN). Odor recognition follows a combinatorial coding scheme, where one OR can be activated by a set of odorants and one odorant can activate a combination of ORs. Through such combinatorial coding, organisms can detect and discriminate between a myriad of volatile odor molecules. Thus, an odor at a given concentration can be described by an activation pattern of ORs, which is specific to each odor. In that sense, cracking the mechanisms that the brain uses to perceive odor requires the understanding odorant-OR interactions. This is why the olfaction community is committed to "de-orphanize" these receptors. Conventional in vitro systems used to identify odorant-OR interactions have utilized incubating cell media with odorant, which is distinct from the natural detection of odors via vapor odorants dissolution into nasal mucosa before interacting with ORs. Here, we describe a new method that allows for real-time monitoring of OR activation via vapor-phase odorants. Our method relies on measuring cAMP release by luminescence using the Glosensor assay. It bridges current gaps between in vivo and in vitro approaches and provides a basis for a biomimetic volatile chemical sensor.

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Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

April 23, 2019

Issue

146

Location

United States

Related Subject Headings

  • Volatilization
  • Receptors, Odorant
  • Olfactory Perception
  • Odorants
  • Nasal Mucosa
  • Humans
  • Cyclic AMP
  • Cell Line
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences
 

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de March, C. A., Fukutani, Y., Vihani, A., Kida, H., & Matsunami, H. (2019). Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase. J Vis Exp, (146). https://doi.org/10.3791/59446
March, Claire A. de, Yosuke Fukutani, Aashutosh Vihani, Hitoshi Kida, and Hiroaki Matsunami. “Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase.J Vis Exp, no. 146 (April 23, 2019). https://doi.org/10.3791/59446.
de March CA, Fukutani Y, Vihani A, Kida H, Matsunami H. Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase. J Vis Exp. 2019 Apr 23;(146).
de March, Claire A., et al. “Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase.J Vis Exp, no. 146, Apr. 2019. Pubmed, doi:10.3791/59446.
de March CA, Fukutani Y, Vihani A, Kida H, Matsunami H. Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase. J Vis Exp. 2019 Apr 23;(146).

Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

April 23, 2019

Issue

146

Location

United States

Related Subject Headings

  • Volatilization
  • Receptors, Odorant
  • Olfactory Perception
  • Odorants
  • Nasal Mucosa
  • Humans
  • Cyclic AMP
  • Cell Line
  • 3101 Biochemistry and cell biology
  • 1702 Cognitive Sciences