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In the rat brain, gamma-hydroxybutyric-acid (GHB) increases the concentrations of 3alpha-hydroxy,5alpha-pregnan-20-one (allopregnanolone, 3alpha,5alpha-THP) and 3alpha,21-dihydroxy,5alpha-pregnan-20-one (allotetrahydrodeoxycorticosterone/3alpha,5alphaTHDOC), two neurosteroids acting as positive allosteric modulators of gamma-aminobutyric acid (GABA)(A) receptors. This study was aimed at assessing whether neurosteroids play a role in GHB-induced loss of righting reflex (LORR). Basal and GHB-stimulated brain concentrations of endogenous 3alpha,5alpha-THP and 3alpha,5alpha-THDOC were analyzed in two rat lines, GHB-sensitive (GHB-S) and GHB-resistant (GHB-R), selectively bred for opposite sensitivity to GHB-induced sedation/hypnosis. Basal neurosteroid concentrations were similar in brain cortex of the two rat lines. However, in male GHB-S rats, administration of GHB (1000 mg/kg, i.p., 30 min) increased brain cortical concentrations of 3alpha,5alpha-THP and 3alpha,5alpha-THDOC 7- and 2.5-fold, respectively, whilst male GHB-R animals displayed only a 4- and 2-fold increase, respectively. In GHB-S rats this increase lasted up to 90 min and declined 180 min following GHB administration, a time course that matches LORR onset and duration. In contrast, in GHB-R rats, which failed to show GHB-induced LORR, brain cortical 3alpha,5alpha-THP and 3alpha,5alpha-THDOC had returned to control values within 90 min. At onset of LORR, a similar increase in brain cortical levels of 3alpha,5alpha-THP and 3alpha,5alpha-THDOC (2-3-fold) was observed in GHB-S female rats and in the few female GHB-R rats that lost the righting reflex after GHB administration, but not in female GHB-R rats failing to show LORR. Sub-hypnotic doses (7.5 and 12.5 mg/kg, i.p.) of pregnanolone, administered 10 min before GHB, dose-dependently facilitated the expression of GHB-induced LORR in GHB-R male rats. These results suggest that the GHB-induced increases of brain 3alpha,5alpha-THP and 3alpha,5alpha-THDOC concentrations are implicated in the eliciting of the sedative/hypnotic action of GHB.

Within the last 20 years, rapid nongenomic actions of steroid hormones have been demonstrated to occur via an interaction with ligand-gated ion channels. For example, the pregnane neurosteroid allopregnanolone (ALLOP) is a potent positive modulator of gamma-aminobutyric acid(A) (GABA(A)) receptors. The physiological significance of fluctuations in endogenous ALLOP levels has been investigated with regard to disease states and the effect of therapeutic agents on ALLOP levels. Because the pharmacological profile of ALLOP is similar to that of ethanol (EtOH), the modulatory effect of pregnane neurosteroids on EtOH dependence and withdrawal will be the focus of this review. Data on the effects of chronic EtOH exposure and withdrawal on pregnane neurosteroid levels, biosynthetic enzymes, and changes in neurosteroid sensitivity will be summarized. Results from genetic animal models indicate that seizure-prone animals have a persistent decrease in endogenous ALLOP levels during EtOH withdrawal in conjunction with tolerance to ALLOP's anticonvulsant effect. Manipulation of endogenous ALLOP levels with finasteride also markedly reduced the severity of chronic EtOH withdrawal. Gene mapping studies provide a hint for an interaction between genes for GABA(A) receptor subunits and the biosynthetic enzyme 5alpha-reductase. Overall, the results are suggestive of a relationship between endogenous pregnane neurosteroid levels and behavioral changes in excitability during EtOH withdrawal, consistent with recent findings in humans. While the findings with ALLOP emphasize the therapeutic potential of neurosteroid treatment during EtOH withdrawal, the gene mapping studies suggest that pregnane neurosteroid biosynthesis may represent a target for therapeutic intervention in the treatment of alcohol dependence.

11 beta,18-Epoxy-2 alpha,3 alpha,18,21-tetrahydroxy-5 alpha,17 alpha- pregnan-20-one (2 alpha-hydroxy-3 alpha,5 alpha-tetrahydro-17-isoaldosterone) and its apo isomer have been identified by high-field NMR studies, supported by thermospray HPLC/MS, to be among the major polar metabolites formed from incubation of aldosterone with rat liver microsomal fraction. Indications that unreduced 2 alpha-hydroxy-aldosterone is also present among the metabolites have still to be confirmed.

Activities of 4-ene-5 alpha-reductase and the metabolism of progesterone were estimated in ovaries of guinea pigs, rats and golden hamsters. In the ovaries of 20-90-day old guinea pigs, the major C21-17-hydroxysteroids and C19-steroids formed from progesterone were 4-ene-3-ketosteroids, such as 17-hydroxy-4-pregnene-3,20-dione and 4-androstene-3,17-dione, and 5 alpha-C21-17-hydroxysteroids and 5 alpha-C19-steroids were not produced. In contrast, the major steroids formed by the ovaries of immature rats and immature hamsters were 5 alpha-steroids, such as 3 alpha, 17-dihydroxy-5 alpha-pregnan-20-one and androsterone, and no 4-ene-3-ketosteroids were found. The 5 alpha-reductase activity (nmol/g tissue/h) in the ovaries of immature hamsters was approx 150 times higher than those in the ovaries of guinea pigs of different ages. In contrast to rats and hamsters, the ovaries of immature guinea pigs do not form a significant amount of 5 alpha-androgens.

3 alpha OH-5 alpha-Pregnan-20-one (allo-THP), a steroid with strong anaesthetic properties, was found to be secreted by the adrenal gland of the rat in quantities similar to those secreted by the rat ovary. From the hypnotic potencies established for this and other endogenous steroids there can be little doubt that the total amount of steroids with anaesthetic properties produced in a female rat are sufficient to exert a depressant action on certain cells of the brain. In rats with intact adrenal glands a positive correlation existed between the adrenal secretion of allo-THP and pregnenolone or progesterone, whereas that between allo-THP and DOC was negative. This could be the result of a competition between the enzymes responsible for the oxidation and reduction of progesterone, the common precursor of allo-THP and DOC. The possibility that allo-THP could have hypotensive actions was suggested.