Characterization of μ, κ, and δ opioid binding in amphibian whole brain tissue homogenates

Leslie C. Newman, Steven S. Sands, David R. Wallace, Craig W. Stevens

Research output: Contribution to journalArticle

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Abstract

Opioid agonists produce analgesia in mammals through the activation of μ, κ, or δ opioid receptors. Previous behavioral and binding studies from our laboratory using an amphibian model suggested that μ, κ, or δ opioid agonists may activate a single type of opioid receptor in the grass frog, Rana pipiens. In the present study, kinetic, saturation, and competitive binding profiles for three opioid radioligands, [3H]DAMGO ([D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin) (μ-selective), [3H]U65953 [(5α, 7α,8β)-(+)-N-methyl-N-[7-(1-pyrrolidinyl) -1-oxaspiro[4.5]dec-8-yl]-benzeneacetamide] (κ-selective), and [3H]DPDPE ([D-Pen2,D-Pen5]-enkephalin) (δ-selective) were determined using frog whole brain homogenates. Kinetic analyses and experimentally derived values from saturation experiments gave affinity constants (KD) in the low nanomolar range. The density of opioid binding sites (Bmax) was 224.4, 118.6, and 268.9 fmol/mg for μ, κ and δ opioid radioligands, respectively. The affinity values did not significantly differ among the three opioid radioligands, but the κ radioligand bound to significantly fewer sites than did the μ or δ radioligands. Ki values for unlabeled μ, κ, and δ competitors, including highly selective opioid antagonists, were consistent with each radioligand selectivity profile, The present data suggest that μ, κ, and δ opioid radioligands bind to distinct opioid receptors in amphibians that are surprisingly similar to those found in mammalian brain.

Original languageEnglish
Pages (from-to)364-370
Number of pages7
JournalJournal of Pharmacology and Experimental Therapeutics
Volume301
Issue number1
DOIs
StatePublished - 6 Apr 2002

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Amphibians
Opioid Analgesics
Brain
Opioid Receptors
D-Penicillamine (2,5)-Enkephalin
Anura
Benzeneacetamides
Ala(2)-MePhe(4)-Gly(5)-enkephalin
Rana pipiens
Competitive Binding
Narcotic Antagonists
Enkephalins
Poaceae
Analgesia
Mammals
Binding Sites

Cite this

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abstract = "Opioid agonists produce analgesia in mammals through the activation of μ, κ, or δ opioid receptors. Previous behavioral and binding studies from our laboratory using an amphibian model suggested that μ, κ, or δ opioid agonists may activate a single type of opioid receptor in the grass frog, Rana pipiens. In the present study, kinetic, saturation, and competitive binding profiles for three opioid radioligands, [3H]DAMGO ([D-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin) (μ-selective), [3H]U65953 [(5α, 7α,8β)-(+)-N-methyl-N-[7-(1-pyrrolidinyl) -1-oxaspiro[4.5]dec-8-yl]-benzeneacetamide] (κ-selective), and [3H]DPDPE ([D-Pen2,D-Pen5]-enkephalin) (δ-selective) were determined using frog whole brain homogenates. Kinetic analyses and experimentally derived values from saturation experiments gave affinity constants (KD) in the low nanomolar range. The density of opioid binding sites (Bmax) was 224.4, 118.6, and 268.9 fmol/mg for μ, κ and δ opioid radioligands, respectively. The affinity values did not significantly differ among the three opioid radioligands, but the κ radioligand bound to significantly fewer sites than did the μ or δ radioligands. Ki values for unlabeled μ, κ, and δ competitors, including highly selective opioid antagonists, were consistent with each radioligand selectivity profile, The present data suggest that μ, κ, and δ opioid radioligands bind to distinct opioid receptors in amphibians that are surprisingly similar to those found in mammalian brain.",
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Characterization of μ, κ, and δ opioid binding in amphibian whole brain tissue homogenates. / Newman, Leslie C.; Sands, Steven S.; Wallace, David R.; Stevens, Craig W.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 301, No. 1, 06.04.2002, p. 364-370.

Research output: Contribution to journalArticle

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