Journal
/
Neuroscience
/
Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay
JoVE Journal
Neuroscience
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Neuroscience
Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay

Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay

DOI:
10.3791/55831-v

•

09:11 min

•

October 02, 2017

•

, , ,
1Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education,Shanghai Jiao Tong University School of Medicine, 2Department of Molecular Genetics and Microbiology,Duke University Medical Center, 3Department of Neurobiology, Duke Institute for Brain Sciences,Duke University Medical Center, 4Institute of Health Science,Chinese Academy of Science/Shanghai Jiao Tong University School of Medicine

Chapters

  • 00:05Title
  • 00:50Plating and Transfection of Hana3A Cells
  • 03:03Transfection of Plasmids
  • 05:24Stimulation and Measuring OR Activity Using the Real-time cAMP Assay
  • 07:27Results: Kinetic Measurements of the Response of OR5AN1 to Muscone
  • 08:23Conclusion

Summary

Automatic Translation

Automatic Translation

Characterizing the function of odorant receptors serves an indispensable part in the deorphanization process. We describe a method to measure the activation of odorant receptors in real time using a cAMP assay.

Tags

Keywords: Odorant ReceptorCAMP AssayLive-cell MeasurementHigh-throughput ScreeningHeterologous Expression SystemHana3A CellsTransfectionOdorant-to-receptor ActivationPhase-contrast MicroscopyCell CultureTrypsinizationCell Counting96-well PlateIncubationPlasmid ConstructsTransfection Mixture

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/55831?article_type=v

Odorant-induced Responses Recorded from Olfactory Receptor Neurons using the Suction Pipette Technique
check_url/55831?article_type=v

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex
check_url/55831?article_type=v

High-throughput Analysis of Mammalian Olfactory Receptors: Measurement of Receptor Activation via Luciferase Activity
check_url/55831?article_type=v

Real-time Electrophysiology: Using Closed-loop Protocols to Probe Neuronal Dynamics and Beyond
check_url/55831?article_type=v

Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor
check_url/55831?article_type=v

A Protocol for the Administration of Real-Time fMRI Neurofeedback Training
check_url/55831?article_type=v

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells
check_url/55831?article_type=v

Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization
check_url/55831?article_type=v

SwarmSight: Real-time Tracking of Insect Antenna Movements and Proboscis Extension Reflex Using a Common Preparation and Conventional Hardware
check_url/55831?article_type=v

A High-content Assay for Monitoring AMPA Receptor Trafficking

Read Article