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Selective loss of delta opioid analgesia and binding by antisense oligodeoxynucleotides to a delta opioid receptor
K M Standifer1, C C Chien, C Wahlestedt
1Cotzias Laboratory of Neuro-Oncology, Memorial Sloan-Kettering Cancer Center, New York, New York.
Abstract:
Antisense oligodeoxynucleotides (18-20 bases) to a cloned delta opioid receptor (DOR-1) lower delta binding in NG108-15 cells by 40%-50%. Changing 4 bases to generate a mismatch antisense oligodeoxynucleotide or mixing the corresponding sense and antisense oligodeoxynucleotides prior to treatment of the cells eliminates the inhibition of binding, confirming the specificity of the response. In vivo, an antisense oligodeoxynucleotide to DOR-1 given intrathecally lowers delta, but not mu or kappa 1 spinal analgesia. The mismatch antisense oligodeoxynucleotide is inactive. Delta analgesic sensitivity gradually returns by 5 days after the last antisense treatment, indicating the lack of irreversible damage or toxicity. These studies demonstrate that DOR-1 mediates delta analgesia at the level of the spinal cord and confirm at the molecular level traditional pharmacological studies implying distinct receptor mechanisms for delta, mu, and kappa 1 analgesia. The use of antisense approaches may prove valuable in understanding the receptors mediating opioid pharmacology.
Insights
Antisense oligodeoxynucleotides targeting the delta opioid receptor (DOR-1) effectively reduce delta analgesia in the spinal cord. This specificity confirms DOR-1
Area of Science:
- Neuroscience
- Pharmacology
- Molecular Biology
Background:
- Opioid receptors, including delta opioid receptor (DOR-1), play crucial roles in pain modulation.
- Traditional pharmacological studies suggest distinct receptor subtypes mediate different types of analgesia (delta, mu, kappa).
Purpose of the Study:
- To investigate the role of DOR-1 in mediating spinal delta analgesia using antisense oligodeoxynucleotides.
- To confirm the specificity of DOR-1 in delta analgesia at the molecular level.
Main Methods:
- Administration of antisense oligodeoxynucleotides targeting DOR-1 to NG108-15 cells and in vivo (intrathecal).
- Assessment of delta opioid receptor binding and spinal analgesia.
- Use of mismatch antisense and sense oligodeoxynucleotides to confirm specificity.
Main Results:
- Antisense oligodeoxynucleotides to DOR-1 reduced delta binding in cells by 40-50%.
- Intrathecal administration of DOR-1 antisense oligodeoxynucleotides lowered spinal delta analgesia but not mu or kappa 1.
- Analgesic sensitivity returned within 5 days, indicating no irreversible toxicity.
Conclusions:
- DOR-1 mediates delta analgesia at the spinal cord level.
- Antisense technology confirms distinct receptor mechanisms for different opioid analgesias.
- Antisense approaches are valuable tools for studying opioid receptor pharmacology.