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Related Experiment Videos

mu opiate receptor: cDNA cloning and expression

J B Wang1, Y Imai, C M Eppler

  • 1Molecular Neurobiology Branch, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD.

Proceedings of the National Academy of Sciences of the United States of America
|November 1, 1993
PubMed
Summary

Researchers identified a novel mu opiate receptor in rat brains. This receptor strongly binds to morphine and other pain-relieving opioid drugs and peptides.

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Area of Science:

  • Neuroscience
  • Pharmacology
  • Molecular Biology

Background:

  • Opiate receptors are crucial for pain perception and analgesia.
  • Existing opiate receptor subtypes (delta and kappa) have distinct binding profiles.
  • The specific molecular identity and function of the mu opiate receptor remained to be fully elucidated.

Purpose of the Study:

  • To identify and characterize the cDNA encoding the mu opiate receptor.
  • To determine the binding affinities and specificities of the expressed receptor.
  • To understand the role of the mu opiate receptor in recognizing analgesic and euphoric compounds.

Main Methods:

  • Polymerase chain reaction (PCR) and cDNA homology screening were used to identify the target cDNA.
  • Northern blot analysis was performed to detect mRNA expression.

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  • COS cells were transfected and expressed the receptor for functional binding assays.

    • Main Results:

    • A 2.1-kb rat brain cDNA was identified, showing 63% identity to delta and kappa opiate receptor sequences.
    • The cDNA recognized a 10.5-kb mRNA predominantly expressed in thalamic neurons.
    • Expressed receptor exhibited naloxonazine-sensitive binding, recognizing mu-selective ligands (morphine, DAMGO, DADLE) with high affinity.

    Conclusions:

    • A novel mu opiate receptor has been molecularly cloned and functionally expressed.
    • This receptor avidly binds analgesic and euphoric opiate drugs and opioid peptides.
    • The findings provide a molecular basis for understanding the actions of clinically important opioid medications.