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[Expression of human mu-opioid receptor cDNA in CHO cell]
Wei Liu1, Hai-Qing Duan, Shu-Qin Li
1Beijing Institute of Biotechnology, Beijing 100071, China.
Abstract:
Opioid receptor, is classified into three subtypes, mu, kappa and delta, with the mu-type receptor plays important roles in opioid analgesia and opioid addiction. The cDNA encoding mu-type receptor was obtained by RT-PCR from human brain RNA and was cloned into pcDNA3.1(+). The resultant recombinant plasmid pcDNAMORs were transfected into CHO cells by liposome. After PCR identification, the positive clone were treated with agonist and antiagonist were tested for their competence of signal transduction. CHO cells that contained mu-opioid receptor in the expression vector pcDNA3.1(+) acquired naloxone-blockable high-affinity specific binding of morphine and DAMGO. The concentration of cAMP in CHO cells transfected with pcDNAMOR was reduced after binding to morphine and DAMGO, and increased after binding naloxone. These results indicate that the mu-type receptor expreesd on the CHO cell has similar biological property as the nature receptor. The availability of these specific cell lines will facilitate the drug development and promote our understanding the mechanism underlying opiate addiction.
Insights
Researchers created a cell model expressing the mu-opioid receptor for studying opioid effects. This model accurately mimics natural receptor function, aiding drug development for pain and addiction.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Context:
- The mu-opioid receptor is crucial for opioid analgesia and addiction.
- Understanding its function is key to developing targeted therapies.
Purpose:
- To create a functional cell model of the mu-opioid receptor.
- To validate its biological properties and signaling pathways.
Summary:
- Human mu-opioid receptor cDNA was cloned and expressed in CHO cells.
- The resulting cells demonstrated specific binding to opioid ligands and modulated cAMP levels, similar to native receptors.
- Naloxone effectively blocked these opioid-induced effects.
Impact:
- Provides a valuable tool for studying mu-opioid receptor pharmacology.
- Facilitates the development of novel analgesics and addiction treatments.
- Enhances understanding of the molecular mechanisms underlying opioid action and addiction.
