title: Opiate binding in calf thalamic membranes: a selective mu 1 binding assay.
abstract: In the present study, we examined the binding of [3H][D-Ala2,D-Leu5]enkephalin ([ 3H]DADL) to bovine thalamic membranes. Scatchard plots were linear with a KD of 0.7 nM. However, competition experiments suggested binding heterogeneity. Approximately 20% of [3H]DADL binding was easily inhibited by [D-Pen2,D-Pen5]enkephalin (DPDPE) and was insensitive to morphine, implying labeling of delta receptors. The remaining 80% of binding was quite sensitive to both morphine and [D-Ala2,MePhe4,Gly(ol)5]enkephalin (DAGO) and insensitive to DPDPE, consistent with a mu receptor. However, this binding did not correspond to classical morphine-selective mu receptors. Unlike morphine-selective receptors, this binding had similar affinities for morphine, DAGO, DADL and [D-Ser2,Leu5]enkephalin-Thr6 (DSLET). In addition, it was far more sensitive to naloxonazine's wash-resistant inhibition and magnesium-induced enhancement of binding than either the morphine-selective (mu 2) or delta sites. [3H]DSLET binding yielded results very similar to those using [3H]DADL. In conclusion, approximately 80% of [3H]DADL binding in thalamus corresponds to a mu receptor distinct from the classical morphine-selective site. Based upon the results of our studies, we feel that this binding represents mu 1 receptors. DPDPE (10 nM) can effectively inhibit the binding of [3H]DADL to delta receptors, leaving a relatively homogeneous labeling of mu 1 sites. The availability of this selective binding assay should facilitate additional studies of mu 1 receptors.
mesh_terms: D000818:Animals; D002417:Cattle; D004399:Dynorphins / Q000378:metabolism; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D020881:Enkephalin, D-Penicillamine (2,5)-; D004743:Enkephalin, Leucine / Q000031:analogs & derivatives / Q000378:metabolism; D016308:Enkephalin, Leucine-2-Alanine; D004745:Enkephalins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D009270:Naloxone / Q000031:analogs & derivatives / Q000494:pharmacology; D009842:Oligopeptides / Q000378:metabolism; D011957:Receptors, Opioid / Q000032:analysis / Q000378:metabolism*; D017465:Receptors, Opioid, delta; D017450:Receptors, Opioid, mu; D013788:Thalamus / Q000378:metabolism*
chemical_list: D004745:Enkephalins; D009842:Oligopeptides; D011957:Receptors, Opioid; D017465:Receptors, Opioid, delta; D017450:Receptors, Opioid, mu; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D009270:Naloxone; D004743:Enkephalin, Leucine; D016308:Enkephalin, Leucine-2-Alanine; D004399:Dynorphins; C034318:enkephalin, Ser(2), Leu(5), Thr(6)-; C034643:naloxonazine; D020881:Enkephalin, D-Penicillamine (2,5)-