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The binding motif to the androgen receptor by DAX-1 is still to be elucidated. Another corepressor, FoxG1, appears to be a likely candidate for interaction with the androgen receptor in vivo but studies are necessary to prove whether this is the case (Obendorf et al. Anabolic steroids bind to the androgen receptor with different affinities. The order of relative binding affinities in comparison with methyltrienolone, which had the strongest affinity, was nandrolone>methenolone>testosterone>mesterolone; a group which had relatively high and generally similar affinity for the androgen receptor in all three tissues. These investigators did not rank 17-methyltestosterone, but it had a relative binding affinity of 0., top 100 steroid sites. The relative binding with fluoxymesterone, methandienone and stanozolol was much weaker and that with oxymetholone and ethylestrenol was too low to be determined. There is a large discrepancy as to what is known about the in vivo activities of these steroids compared with their in vitro activity, even taking into account possible differences in the bioavailability and clearance of these steroids (not least determined by the affinity to sex hormone-binding globulin in the blood circulation). For example, oxymetholone and stanozolol have low relative binding affinity compared with 17?-methyltestosterone in the in vitro study, but, conversely, these steroids have a relatively high myotrophic activity compared with the same steroid when administered to the castrated rat (see Table 1 ). Furthermore, Feldkoren and Andersson (2005) found that stanozolol and methandienone have significantly lower binding affinities compared with testosterone but all three steroids were potent activators in a cell-based androgen receptor-dependent transactivation assay. Clearly, the degree of physical binding to the androgen receptor, as measured by ligand-binding assays, does not fully explain the biological activity of anabolic steroids. Distinct target gene expression profiles due to androgen receptor activation by structurally different androgens has also been reported (Holterhus et al. The model used was three structurally different androgen promoter constructs in co-transfected Chinese hamster ovary cells. All the steroids proved to be potent activators of the androgen receptor, but the anabolic steroids and the testosterone precursors showed characteristic promoter activation profiles distinct from the virilizing androgens. The assumption is that the specific ligand-induced conformation determines how the hormone receptor complex can specifically interact with coregulators and neighbouring transcription factors and also that the transactivation capability depends on the structure of the response element. Even though many co-activators have been identified as enhancing the ligand-induced transcriptional activity of the androgen receptor, their relative importance with respect to particular cell types and tissues is unclear (Heinlein and Chang, 2002b). The emerging knowledge concerning androgen receptor interaction with its coregulators in different tissues clearly has relevance to understanding how anabolic steroids exert their actions and will give further insight into how favourable anabolic’androgenic dissociation may be achieved. Most recently, in vitro bioassays have been employed to determine that the designer anabolic steroid THG is indeed a potent androgen. The authors make the important point that it is not known whether yeast-based systems express the repertoire of coregulators that is present in mammalian androgen-responsive tissues. These investigators found that THG and DHT modulated in a similar fashion 671 genes in the mouse levator ani muscle, 95 genes in the gastrocnemius muscle and 939 genes in the prostate. The use of in vitro assays based on androgen receptor expression, as described above, can help to assess whether future designer steroids have anabolic’androgenic activity, and can help to minimize in vivo experiments. These approaches can provide useful evidence to government agencies involved in the regulation of drugs to protect public health. Moreover, the employment of such assays should be of particular benefit to sporting authorities to help stifle legal challenges based on the premise that new designer steroids have unproven anabolic activity and thus should not be subject to doping control and the penalties associated with their administration. It is accepted that the administration of anabolic steroids to healthy women and children has an anabolic effect, and that with the virilizing effects, there is a gain in muscle mass and strength. However, for many years, it was difficult to prove conclusively that the administration of these steroids had a myotrophic effect in healthy young sportsmen, as discussed by Ryan (1976) (see the section ‘Anabolic steroids as performance enhancers in sport’). Around that period, an interesting but speculative biochemical explanation for this difference in response between the sexes was that due to the exposure to testosterone during puberty in men, there is a downregulation of receptors (decrease in responsiveness of receptors often followed by decrease in numbers) in the skeletal muscle and that the androgen receptor population is then saturated with testosterone in the adult, so that no further response can be induced by pharmacological doses of androgens (Wilson, 1988).
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