Topography and characteristics of specific binding sites for non-opioid gamma-type endorphins in the rat brain as studied by autoradiography with [35S]Met-desenkephalin-gamma-endorphin.
An in vitro autoradiographic study was performed to characterize specific rat brain binding sites for non-opioid neuroleptic-like gamma-type endorphins, using [35S]Met-des-enkephalin-gamma-endorphin ([35S]Met-DE gamma E; [35]S-beta-endorphins(5-17)) with high specific activity as radioligand. The binding sites appeared to be confined to rat forebrain regions, e.g., orbital cortex, frontal cortex, cingulate cortex, piriform cortex, nucleus accumbens, amygdala, mediodorsal nucleus of the thalamus and arcuate and periventricular nuclei of the hypothalamus. These regions are part of the mesocorticolimbic feedback circuit. Densitometric analysis of the autoradiographs revealed that the density of the binding sites was highest in the mediodorsal nucleus of the thalamus and the amygdala. Concentration-dependent displacement of [35S]Met-DE gamma E (500 pM) with DE gamma E yielded an IC50 of 0.6 nM whereas DE alpha E (beta-endorphin(6-16)) had an IC50 of 210 nM. Various endorphins, sharing the gamma-endorphin C terminus, displaced [35S]Met-DE gamma E to the same extent as non-labelled DE gamma E (at 10(-6) M) whereas non-endorphin peptides did not show displacing capacity. Possible relationships of the binding sites with opioid receptors were investigated. DAMGO (mu) and DPDPE (delta) displaced [35S]Met-DE gamma E to some extent at 10(-6) M whereas U69,593 (kappa) was inactive, suggesting that the binding sites for gamma-type endorphins may resemble mu- and delta-opioid receptors in some aspects. Similarly, relationships with dopamine receptors were investigated. Haloperidol partially displaced [35S]Met-DE gamma E whereas sulpiride, SKF38,393 and 3-PPP at 10(-6) M did not induce significant displacement. Thus, binding sites are distinct from dopamine receptors.(ABSTRACT TRUNCATED AT 250 WORDS)
Ronken
E
E
Rudolf Magnus Institute, Department of Medical Pharmacology, University of Utrecht, The Netherlands.
Wiegant
V M
VM
Kaspersen
F M
FM
van Nispen
J W
JW
de Boer
T
T
Bruning
H W
HW
Rust
C J
CJ
Tonnaer
J A
JA
D000679:Amygdala / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D001345:Autoradiography; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003720:Densitometry; D004723:Endorphins / Q000378:metabolism*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D011954:Receptors, Dopamine / Q000378:metabolism; D011957:Receptors, Opioid / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013462:Sulfur Radioisotopes; D001615:beta-Endorphin / Q000276:immunology
D000911:Antibodies, Monoclonal; D004723:Endorphins; D011954:Receptors, Dopamine; D011957:Receptors, Opioid; D013462:Sulfur Radioisotopes; D001615:beta-Endorphin; C033558:des-enkephalin-gamma-endorphin