RU 3117 a steroidal compound with high affinity for sigma sites in rat testis membranes.
RU 3117 belongs to a new series of steroids which exhibited a high relative binding affinity (RBA) for (+)[3H]PPP sites in rat testis membranes; its RBA was about 40 times higher than that of progesterone. Furthermore, it is devoid of any binding to classical steroid receptors; therefore in order to study its binding parameters on rat testis membranes it was tritiated. [3H]RU 3117 bound at least two distinct sites with Ka values of 0.4 +/- 0.06 x 10(9) M(-1) and 1.3 +/- 0.2 x 10(7) M(-1). Using this marker, competition studies with cold haloperidol showed that a part of this binding was haloperidol-sensitive, whereas another part was haloperidol-resistant. Interestingly, progesterone described as a sigma ligand competes with [3H]RU 3117 binding, with an RBA of 1.6%. When haloperidol was preincubated (250 nM) with rat testis membranes, in order to mask the sigma sites, we observed that DTG (1,3-di-O-tolylguanidine) and haloperidol displayed a very low RBA (< 0.1%) and were not able totally to displace the [3H]RU 3117 binding up to 50 microM. Furthermore, benztropine exhibited a significant RBA of 19% but its displacement curve showed a plateau (500-50,000 nM). These results showed that part of the haloperidol-resistant sites was benztropine sensitive but another part was displaced neither by haloperidol nor by benztropine. The presence of these remaining binding sites was confirmed by preincubating a mixture of haloperidol and benztropine with testis membranes. Under these conditions, [3H]RU 3117 displayed a Ka of 1.0 +/- 0.01 x 10(7) M(-1), and we observed that these sites were recognized, up to now, only by the steroids RU 1968 and RU 54173 which are also devoid of any binding to classical nuclear steroid receptors.
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Centre de Recherches Roussel UCLAF, Romainville, France.
Humbert
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Philibert
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D000818:Animals; D001590:Benztropine / Q000378:metabolism; D001667:Binding, Competitive; D004963:Estrenes / Q000737:chemistry / Q000378:metabolism; D004970:Estrone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D006220:Haloperidol / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008566:Membranes / Q000378:metabolism; D018377:Neurotransmitter Agents / Q000378:metabolism; D011281:Pregnatrienes / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D017480:Receptors, sigma / Q000378:metabolism*; D013256:Steroids / Q000378:metabolism; D013737:Testis / Q000378:metabolism*
C104062:3-(beta-ethylaminoethoxy)-delta-1,3,5-estratriene-17-one; C104061:3-(n-dimethylaminoethyloxy)estra-1,3,5-triene; D004963:Estrenes; D018377:Neurotransmitter Agents; D011281:Pregnatrienes; C104063:RU 1968; D017480:Receptors, sigma; D013256:Steroids; D001590:Benztropine; D004970:Estrone; D006220:Haloperidol