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Neuroscience
실시간 체 외 Odorant 수용 체 활성화는 Odorant 수증기 단계에서의 모니터링
실시간 체 외 Odorant 수용 체 활성화는 Odorant 수증기 단계에서의 모니터링
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Real-time In Vitro Monitoring of Odorant Receptor Activation by an Odorant in the Vapor Phase

실시간 체 외 Odorant 수용 체 활성화는 Odorant 수증기 단계에서의 모니터링

Full Text
7,477 Views
09:53 min
April 23, 2019

DOI: 10.3791/59446-v

Claire A. de March1, Yosuke Fukutani1,2, Aashutosh Vihani1,3, Hitoshi Kida1,4, Hiroaki Matsunami1,3,5,6

1Department of Molecular Genetics and Microbiology,Duke University Medical Center, 2Department of Biotechnology and Life Science,Tokyo University of Agriculture and Technology, 3Department of Neurobiology,Duke University Medical Center, 4Department of Mechanical Systems, Engineering,Tokyo University of Agriculture and Technology, 5Institute of Global Innovation Research,Tokyo University of Agriculture and Technology, 6Duke Institute for Brain Sciences,Duke University

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Overview

This study presents a novel method for real-time in vitro monitoring of odorant receptor activation using vapor phase odorant stimulation. Previous in vitro systems have largely relied on liquid phase stimulation, potentially limiting physiological relevance. The method provides insights into the activation kinetics of odorant receptors and their role in odor perception.

Key Study Components

Area of Science

  • Neuroscience
  • Olfactory biology
  • Cell culture techniques

Background

  • Odorant receptors respond to volatile odorant molecules, crucial for the sense of smell.
  • Existing in vitro methods primarily use liquid phase odorants, which may not accurately mimic natural conditions.
  • This research addresses gaps in current methodologies by allowing for vapor phase stimulation.

Purpose of Study

  • To develop a method that facilitates the detection and discrimination of volatile odors in vitro.
  • To enable real-time monitoring of odorant receptor activation.
  • To bridge the gap between in vitro and in vivo approaches in olfactory research.

Methods Used

  • Utilized cell culture techniques to grow cells in M10 PSF medium at physiological conditions.
  • Involves transfection of cells with specific odorant receptor plasmids to study their activation.
  • Key steps include careful cell preparation, including trypsinization and centrifugation.
  • Measurements of luminescence provide data on receptor activation upon exposure to odorants.

Main Results

  • The method allows for precise monitoring of odorant receptor responses to specific odorants like acetophenone.
  • Findings illustrate the kinetics of odor perception and potential roles of mucosal metabolic enzymes.
  • Successfully demonstrated the capability to detect and discriminate odors in a controlled in vitro setting.

Conclusions

  • This innovative approach enhances the understanding of olfactory receptor dynamics in a more physiologically relevant context.
  • The findings enable further research into the development of odorant receptor-based biosensors.
  • Implications extend to understanding mechanisms of odor detection, which could inform broader olfactory studies.

Frequently Asked Questions

What are the advantages of using vapor phase stimulation?
Vapor phase stimulation mimics natural conditions, providing a more physiologically relevant method to study odorant receptors compared to liquid phase approaches.
How is the biological model implemented in this study?
Cells are cultured in specific media and transfected with plasmids encoding odorant receptors to investigate their responses to volatile stimuli.
What types of data are obtained from this method?
The methodology provides luminescence data indicating the activation levels of odorant receptors in response to odorant exposure.
How can this method be applied to other research areas?
This technique can potentially be adapted for various studies involving sensory receptor activation and biosensor development across biological systems.
What are the key limitations or considerations for this study?
Care must be taken to ensure that experimental conditions, such as temperature and CO2 levels, remain constant to obtain accurate results in receptor activation studies.

순수, odorant 수용 체 odorant 분자 수증기 단계에서 흡입에 의해 활성화 됩니다. 그러나, 대부분의 생체 외에서 시스템 활용 액체 단계 odorant 자극. 여기, 선물이 odorant 자극 기상에 따라 odorant 수용 체 활성화의 실시간 생체 외에서 모니터링 수 방법.

우리의 프로토콜은 냄새 수용체 패널로 휘발성 냄새의 검출 및 차별을 측정 할 수 있습니다. 이 방법은 악취 수용체 기반 바이오 센서를 개발할 가능성을 열어줍니다. 이 기술은 증기 상에서 악취 분자에 악취 수용체 활성화의 실시간 체외 모니터링을 할 수 있습니다.

그것은 체외와 생체 내 접근 사이의 간격을 채웁니다. 이 방법은 점막 대사 효소의 역할을 포함하여 악취 지각으로 이끌어 내는 사건의 운동학을 이해하는 데 이용될 수 있습니다. 시작하려면 원고에 따라 M10 및 M10 PSF를 준비하십시오.

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