Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Letter to the Editor: First Experiences with Amyloid-Related Imaging Abnormalities - Yet without Imaging that Can Rule Out Cerebral Injury or Monitor Efficacy of Recommended Management.

The journal of prevention of Alzheimer's disease·2024
Same author

MRI Monitoring of Anti-Alzheimer Therapy Amyloid-Related Imaging Abnormalities: Due Diligence or Overkill?

AJNR. American journal of neuroradiology·2022
Same author

Detection of pyrazinamide resistance of Mycobacterium tuberculosis using nicotinamide as a surrogate.

Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases·2017
Same author

Hepatic cyclooxygenase-2 overexpression induced spontaneous hepatocellular carcinoma formation in mice.

Oncogene·2017
Same author

FAM83A is amplified and promotes cancer stem cell-like traits and chemoresistance in pancreatic cancer.

Oncogenesis·2017
Same author

Epigenomics alternations and dynamic transcriptional changes in responses to 5-fluorouracil stimulation reveal mechanisms of acquired drug resistance of colorectal cancer cells.

The pharmacogenomics journal·2017

Related Experiment Video

Updated: Jul 19, 2026

Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration
08:47

Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration

Published on: February 10, 2012

Noninvasive, quantitative imaging in living animals of a mutant dopamine D2 receptor reporter gene in which ligand

Q Liang1, N Satyamurthy, J R Barrio

  • 1Crump Institute for Biological Imaging, UCLA School of Medicine, Los Angeles, CA 90095-1570, USA.

Gene Therapy
|October 11, 2001
PubMed
Summary

A mutated dopamine D2 receptor (D2R80A) functions as an effective in vivo PET reporter gene. This D2R80A mutant maintains ligand binding while uncoupling from cAMP modulation, enhancing reporter gene utility.

More Related Videos

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models
10:04

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models

Published on: March 13, 2018

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology
07:25

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Published on: March 27, 2018

Related Experiment Videos

Last Updated: Jul 19, 2026

Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration
08:47

Spectral Confocal Imaging of Fluorescently tagged Nicotinic Receptors in Knock-in Mice with Chronic Nicotine Administration

Published on: February 10, 2012

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models
10:04

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models

Published on: March 13, 2018

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology
07:25

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Published on: March 27, 2018

Area of Science:

  • Molecular Biology
  • Neuroscience
  • Biomedical Imaging

Background:

  • The dopamine D2 receptor (D2R) is utilized as an in vivo reporter gene in adenoviral systems and tumor xenografts.
  • Positron emission tomography (PET) enables non-invasive imaging of D2R expression via ligands like FESP.
  • Dopamine binding to D2R can affect cyclic AMP (cAMP) levels, potentially interfering with reporter gene function.

Purpose of the Study:

  • To develop an improved D2R-based reporter gene by uncoupling ligand binding from Gi-protein-mediated cAMP inhibition.
  • To assess the functionality of D2R mutants (D2R80A and D2R194A) in maintaining ligand binding and reporter gene capabilities.

Main Methods:

  • Site-directed mutagenesis was used to create D2R mutants (Asp80 to Alanine, D2R80A; Ser194 to Alanine, D2R194A).
  • Ligand binding assays ([3H]spiperone) and cAMP accumulation assays (forskolin-stimulated) were performed in transfected and infected cells.
  • In vivo PET imaging was conducted using FESP in animals injected with adenoviruses expressing wild-type D2R or D2R80A.

Main Results:

  • The D2R80A mutation completely abolished dopamine's ability to suppress cAMP levels, while D2R194A only partially reduced this effect.
  • Both wild-type D2R and D2R80A showed equivalent [3H]spiperone binding in vitro.
  • In vivo, hepatic FESP sequestration was comparable between adenoviruses expressing D2R and D2R80A, indicating similar reporter gene expression and accessibility.

Conclusions:

  • The D2R80A mutant is a fully functional PET reporter gene, as it retains ligand-binding capacity without modulating cAMP levels.
  • Uncoupling ligand binding from cAMP modulation optimizes the D2R for in vivo reporter gene applications.
  • D2R80A offers a superior tool for non-invasive, quantitative imaging in biomedical research.