The present study was carried out to determine steroid biosynthesis from cholesterol in the brain of adult male Japanese quails. As an initial step of the experiments, the concentrations of pregnenolone, dehydroepiandrosterone and their sulfate esters in the brain and plasma extracts were measured by specific radioimmunoassays (RIAs). To exclude the possibility that these steroids in the brain are derived from peripheral steroidogenic glands, hypophysectomized and sham-operated birds were used in this experiment. The pregnenolone concentration was much larger in the brain than in the plasma in these two groups. Hypophysectomy led to decreases in the plasma and brain pregnenolone concentrations, but the change in the brain was less pronounced than that in the plasma. Although pregnenolone sulfate ester, dehydroepiandrosterone and its sulfate ester were also detected in brain extracts, those levels were low in both hypophysectomized and sham-operated birds. The biochemical demonstration of cholesterol metabolism was further conducted in intact mitochondria. When mitochondrial fractions obtained from the whole brain were incubated with cholesterol at 37°C, the pregnenolone level in mitochondria increased as a function of incubation time. Finally, Western immunoblot analysis using a purified IgG with polyclonal antibodies against the purified bovine adrenal cytochrome P450scc was performed after SDS-gel electrophoresis of homogenates of the hypothalamus and preoptic area. A protein recognized the antibody as a band of electrophoretic mobility in the proximity of reference bovine P450scc. These results suggest that the brain produces pregnenolone from cholesterol in adult male Japanese quails. Most accumulation of pregnenolone in the brain may be independent of the peripheral endocrine gland system.
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