Mass spectrometric assay and physiological-pharmacological activity of androgenic neurosteroids.
Steroid hormones play a key role in the pathophysiology of several brain disorders. Testosterone modulates neuronal excitability, but the underlying mechanisms are obscure. There is emerging evidence that testosterone-derived "androgenic neurosteroids", 3alpha-androstanediol and 17beta-estradiol, mediate the testosterone effects on neural excitability and seizure susceptibility. Testosterone undergoes metabolism to neurosteroids via two distinct pathways. Aromatization of the A-ring converts testosterone into 17beta-estradiol. Reduction of testosterone by 5alpha-reductase generates 5alpha-dihydrotestosterone, which is then converted to 3alpha-androstanediol, a powerful GABA(A) receptor-modulating neurosteroid with anticonvulsant properties. Although the 3alpha-androstanediol is an emerging neurosteroid in the brain, there is no specific and sensitive assay for determination of 3alpha-androstanediol in biological samples. This article describes the development and validation of mass spectrometric assay of 3alpha-androstanediol, and the molecular mechanisms underlying the testosterone modulation of seizure susceptibility. A liquid chromatography-tandem mass spectrometry assay to measure 3alpha-androstanediol is validated with excellent linearity, specificity, sensitivity, and reproducibility. Testosterone modulation of seizure susceptibility is demonstrated to occur through its conversion to neurosteroids with "anticonvulsant" and "proconvulsant" actions and hence the net effect of testosterone on neural excitability and seizure activity depends on the levels of distinct testosterone metabolites. The proconvulsant effect of testosterone is associated with increases in plasma 17beta-estradiol concentrations. The 5alpha-reduced metabolites of testosterone, 5alpha-dihydrotestosterone and 3alpha-androstanediol, had powerful anticonvulsant activity. Overall, the testosterone-derived neurosteroids 3alpha-androstanediol and 17beta-estradiol could contribute to the net cellular actions of testosterone in the brain. Because 3alpha-androstanediol is a potent positive allosteric modulator of GABA(A) receptors, it could serve as an endogenous neuromodulator of neuronal excitability in men. The 3alpha-androstanediol assay is an important tool in this area because of the growing interest in the potential to use adjuvant aromatase inhibitor therapy to improve treatment of epilepsy.
Reddy
Doodipala S
DS
North Carolina State University, Department of Molecular Biomedical Sciences, Raleigh, NC 27606, USA. samba_reddy@ncsu.edu
D000728:Androgens / Q000032:analysis* / Q000494:pharmacology*; D015113:Androstane-3,17-diol / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D047072:Aromatase Inhibitors / Q000494:pharmacology; D004967:Estrogens / Q000502:physiology; D006801:Humans; D013058:Mass Spectrometry; D018377:Neurotransmitter Agents / Q000032:analysis* / Q000096:biosynthesis / Q000494:pharmacology*; D011963:Receptors, GABA-A / Q000502:physiology; D013256:Steroids / Q000032:analysis* / Q000096:biosynthesis / Q000494:pharmacology*; D013739:Testosterone / Q000378:metabolism
D000728:Androgens; D047072:Aromatase Inhibitors; D004967:Estrogens; D018377:Neurotransmitter Agents; D011963:Receptors, GABA-A; D013256:Steroids; D015113:Androstane-3,17-diol; C028895:androstane-3,17-diol glucuronide; D013739:Testosterone