Difference in luteal and placental P45017α: substrate preference and hormonal regulation in the rat

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Abstract
The purpose of this study was to assess the substrate specificity of P45017α in both the corpus luteum and placenta of pregnant rats, and to analyse the site at which LH/human chorionic gonadotrophin (hCG) regulates the activities of this enzyme. To distinguish the substrate preference, placentas and corpora lutea were obtained from rats on day 15 of pregnancy. Tissues were homogenized and the 10 000 g supernatants incubated in the presence of equimolar concentrations of [14C]progesterone and [3H]17α-hydroxyprogesterone as substrate with either NADH or NADPH as cofactors for 2, 8, 16 and 24 min. The labelling pattern of both 17α-hydroxyprogesterone and testosterone indicated that the corpus luteum produced testosterone preferentially from progesterone, whereas the placenta principally used 17α-hydroxyprogesterone and synthesized six times as much testosterone from 17α-hydroxyprogesterone than from progesterone. Addition of either NADPH or NADH as cofactors had no effect on substrate preference. The products of the two enzymatic activities were identified by recrystallization to constant 14C/3H ratios. The ratio of 14C/3H in testosterone produced by the corpus luteum was 16-fold higher than in that produced by the placenta. To explore which of the two activities of P45017α is regulated by the gonadotrophin, rats were treated with either 1·5 IU hCG or vehicle between days 13 and 15 of pregnancy. Hydroxylase and lyase activities were determined on day 15 after incubation for 2,8,16 or 24 min in the presence of either NADH or NADPH. Administration of hCG significantly inhibited NADH-dependent 17α-hydroxylase in the placenta at each time-point studied. The inhibition reached 69% at 24 min. Human chorionic gonadotrophin did not affect the NADPH-dependent 17α-hydroxylase and had only a slight inhibitory effect on both NADH- and NADPH-dependent 17,20-lyase activities in the placenta. In contrast to its effect on the placenta, hCG stimulated both NADH- and NADPH-linked 17,20-lyase activities but had no measurable effect on 17α-hydroxylase activities in the corpus luteum. In summary, the results of the present investigation have revealed a significant difference in the behaviour of 17α-hydroxylase/17,20-lyase activities in the placenta and corpus luteum. The substrate preference and the control of both enzyme activities by LH/hCG differs dramatically. J. Endocr. (1987) 115, 387–393