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Binding of the steroidal pheromone 17 alpha 20 beta dihydroxy 4 pregnen 3 one to goldfish carassius auratus olfactory epithelium membrane preparations



Binding of the steroidal pheromone 17 alpha 20 beta dihydroxy 4 pregnen 3 one to goldfish carassius auratus olfactory epithelium membrane preparations



Chemical Senses 16(2): 143-154



The binding of the steroidal pheromone 17.alpha.,20.beta.-dihydroxy-4-pregnen-3-one (17.alpha.,20.beta.-P) to membranes prepared from the goldfish olfactory epithelium was investigated using an in vitro radioreceptor assay. Maximum specific binding occurred after a 60-min incubation at 4.degree. C. Binding increased linearly with increasing amounts of tissue. 17.alpha.,20.beta.-P binding saturated at a ligand concentration of approximately 4.0 nM, matching well with data obtained from electrophysiological recordings of the in situ sensitivity of the olfactory epithelium to 17.alpha.,20.beta.-P. Scatchard analysis yielded a single class (r = 0.95) of high-affinity (Kd = 1.02 .+-. 0.06 nM), low-capacity (Bmax = 1.38 .+-. 0.24 pmol/mg protein) receptors. Olfactory epithelial membranes were highly enriched in 17.alpha.,20.beta.-P receptors; binding was 10 to 20 times higher than in membranes prepared from gut, liver or brain. The specificity of binding was not absolute. Progestins (progesterone, 17.alpha.-hydroxyprogesterone) and androgens (adrostenedione, testosterone) competed with 17.alpha.,20.beta.-P for binding; however, these steroids were less potent at displacing radiolabeled 17.alpha.,20.beta.-P than was 17.alpha.,20.beta.-P itself. Estrogens and glucocorticoids did not displace 17.alpha.,20.beta.-P in this preparation. These results are consistent with respective abilities of these compounds to elicit electro-olfactogram activity, and confirm the presence of membrane-bound steroidal receptors in olfactory tissue.

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