Abstract
Identifying neurons that have functional opioid receptors is fundamental for the understanding of the cellular, synaptic and systems actions of opioids. Current techniques are limited to post hoc analyses of fixed tissues. Here we developed a fluorescent probe, naltrexamine-acylimidazole (NAI), to label opioid receptors based on a chemical approach termed 'traceless affinity labeling'. In this approach, a high affinity antagonist naltrexamine is used as the guide molecule for a transferring reaction of acylimidazole at the receptor. This reaction generates a fluorescent dye covalently linked to the receptor while naltrexamine is liberated and leaves the binding site. The labeling induced by this reagent allowed visualization of opioid-sensitive neurons in rat and mouse brains without loss of function of the fluorescently labeled receptors. The ability to locate endogenous receptors in living tissues will aid considerably in establishing the distribution and physiological role of opioid receptors in the CNS of wild type animals.
| Original language | English (US) |
|---|---|
| Journal | eLife |
| Volume | 8 |
| DOIs | |
| State | Published - Oct 7 2019 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Neuroscience(all)
- Immunology and Microbiology(all)
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry. / Arttamangkul, Seksiri; Plazek, Andrew; Platt, Emily J.; Jin, Haihong; Murray, Thomas F.; Birdsong, William T.; Rice, Kenner C.; Farrens, David L.; Williams, John T.
In: eLife, Vol. 8, 07.10.2019.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry
AU - Arttamangkul, Seksiri
AU - Plazek, Andrew
AU - Platt, Emily J.
AU - Jin, Haihong
AU - Murray, Thomas F.
AU - Birdsong, William T.
AU - Rice, Kenner C.
AU - Farrens, David L.
AU - Williams, John T.
PY - 2019/10/7
Y1 - 2019/10/7
N2 - Identifying neurons that have functional opioid receptors is fundamental for the understanding of the cellular, synaptic and systems actions of opioids. Current techniques are limited to post hoc analyses of fixed tissues. Here we developed a fluorescent probe, naltrexamine-acylimidazole (NAI), to label opioid receptors based on a chemical approach termed 'traceless affinity labeling'. In this approach, a high affinity antagonist naltrexamine is used as the guide molecule for a transferring reaction of acylimidazole at the receptor. This reaction generates a fluorescent dye covalently linked to the receptor while naltrexamine is liberated and leaves the binding site. The labeling induced by this reagent allowed visualization of opioid-sensitive neurons in rat and mouse brains without loss of function of the fluorescently labeled receptors. The ability to locate endogenous receptors in living tissues will aid considerably in establishing the distribution and physiological role of opioid receptors in the CNS of wild type animals.
AB - Identifying neurons that have functional opioid receptors is fundamental for the understanding of the cellular, synaptic and systems actions of opioids. Current techniques are limited to post hoc analyses of fixed tissues. Here we developed a fluorescent probe, naltrexamine-acylimidazole (NAI), to label opioid receptors based on a chemical approach termed 'traceless affinity labeling'. In this approach, a high affinity antagonist naltrexamine is used as the guide molecule for a transferring reaction of acylimidazole at the receptor. This reaction generates a fluorescent dye covalently linked to the receptor while naltrexamine is liberated and leaves the binding site. The labeling induced by this reagent allowed visualization of opioid-sensitive neurons in rat and mouse brains without loss of function of the fluorescently labeled receptors. The ability to locate endogenous receptors in living tissues will aid considerably in establishing the distribution and physiological role of opioid receptors in the CNS of wild type animals.
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U2 - 10.7554/eLife.49319
DO - 10.7554/eLife.49319
M3 - Article
C2 - 31589142
AN - SCOPUS:85073764978
VL - 8
JO - eLife
JF - eLife
SN - 2050-084X
ER -