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  • Author: P. J. A. Woutersen x
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J. Th. J. Uilenbroek, P. J. A. Woutersen and P. D. M. van der Vaart


Corpora lutea could be identified under the dissection microscope up to 7 days after formation. They were isolated during the oestrous cycle and pseudopregnancy and the progesterone and 20α-OH-progesterone contents were compared with serum values of these steroids. The pattern of progesterone in serum resembled that found in the corpora lutea. However, the pattern of 20α-OH-progesterone concentrations in serum and corpora lutea were different. While 20α-OH-progesterone concentrations in the corpora lutea showed large variations during the cycle, changes in serum concentrations of 20α-OH-progesterone were relatively small. Measurement of hormone concentrations in isolated corpora lutea is therefore a sensitive method for studying corpus luteum activity.

To study whether corpora lutea derived after ovulation of immature follicles showed deficient luteal activity, rats at dioestrus (2 days before pro-oestrus) were induced to ovulate by the injection of 10 IU human chorionic gonadotrophin (hCG) and subsequent luteal activity was studied by measuring hormone concentrations in the corpora lutea on day 5 of pseudopregnancy. Concentrations of progesterone, but not of 20α-OH-progesterone, in corpora lutea derived from follicles induced to ovulate at dioestrusday 1 were significantly lower than those in corpora lutea derived from follicles induced to ovulate at prooestrus. This difference was observed not only when pseudopregnancy was induced by cervical stimulation but also when it was induced by implantation of a pituitary gland under the kidney capsule. However, in the latter case, corpora lutea already present on the day of hCG injection also became activated.

The present experiments demonstrate that by measuring hormone concentrations in isolated corpora lutea changes in luteal activity can be studied effectively. Moreover, it appears that corpora lutea derived from immature follicles contained less progesterone than those derived from fully mature follicles.

Journal of Endocrinology (1989) 120, 325–330

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J. Th. J. Uilenbroek, P. J. A. Woutersen and R. van der Linden

To determine changes in steroidogenesis by rat ovaries during sexual maturation, ovaries obtained at various ages (days 10–35) and at the first pro-oestrus were incubated in the absence or presence of LH and the accumulation of steroids in the medium was measured.

Basal and LH-stimulated oestradiol-17β and testosterone release into the medium, expressed in pmol/4 h per mg ovary, was high at day 10 of age and at first pro-oestrus. Between days 20 and 35 basal oestradiol and testosterone release was low and could not be stimulated by LH. Addition of testosterone to the culture medium increased oestradiol production at all ages studied. Release of progesterone occurred at all ages even in LH-free medium. Incubation in the presence of LH resulted in a dose-dependent increase in progesterone with a maximal response at pro-oestrus. Androsterone and 5α-androstane-3α,17β-diol production in the absence or presence of LH was high during the entire prepuberal period. Production of 5α-reduced androgens in response to LH increased from days 10 to 20 but decreased thereafter. Similarly, 5α-reductase activity, measured in ovarian homogenates, increased from days 10 to 20 but was decreased again by first pro-oestrus. A further decrease in basal and LH-stimulated 5α-reduced androgen production occurred after first ovulation.

These results demonstrated age-related changes in steroid release after in-vitro incubation. At day 10 progesterone can be converted to aromatizable androgens allowing production of oestrogens, while after day 10 progesterone is converted to 5α-reduced C19 steroids. The decrease in 5α-reductase activity correlates with an increase in LH-stimulated testosterone and oestradiol production at the first pro-oestrus.

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Testosterone was measured in plasma pools from male and female fetal guinea-pigs between the ages of 30 and 55 days of pregnancy. Between days 33 and 36 the testosterone concentration in the plasma of males (1·4 ng/ml) was several times higher than that found at other ages or that measured in female fetuses. After infusion of tritiated testosterone for 2 h into pregnant guinea-pigs at day 36 of pregnancy, high levels of testosterone and androstenedione were found in maternal plasma. Nevertheless, tritiated testosterone and androstenedione could hardly be shown in the fetuses. Similar large differences in plasma progesterone levels appeared to exist between the maternal and the fetal circulation. Therefore, only a very small fraction of these steroids can penetrate from the maternal circulation into that of the fetus. This finding might be explained by the large difference in androgen-binding capacity between maternal and fetal plasma, as was shown by equilibrium dialysis.

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J. Th. J. Uilenbroek, H. M. A. Meijs-Roelofs, P. J. A. Woutersen, P. Kramer, W. A. van Cappellen, L. A. Gribling-Hegge and W. J. de Greef


To determine whether the decrease in ovarian 5α-reduced androgen production before first ovulation might be caused by an increase in serum LH, prepuberal female rats were injected at 28–31 days of age with low doses of human chorionic gonadotrophin (hCG) (0·05–0·075 i.u., four times daily). This treatment resulted in ovulation of six to ten ova per rat on day 32 in all animals.

Treatment with hCG resulted in a gradual decrease in ovarian content and production (i.e. content in ovary and medium after 4 h of incubation) of 5α-dihydrotestosterone (DHT) and 5α-androstane-3α,17β-diol. The ovarian content of DHT and the production of 5α-androstane-3α,17β-diol decreased within 24 h after the first injection of hCG. Oestradiol content and production increased between 24 and 48 h after the start of treatment and was maximal on day 31 (day of pro-oestrus).

Activities of 5α-reductase and aromatase were measured in ovarian homogenates obtained on days 29–31. Activity of 5α-reductase in hCG-treated rats was lower than that in control rats on all days studied. Aromatase activity in hCG-treated rats increased between days 29 and 31.

It was concluded that multiple injections of low doses of hCG, which may induce ovulation, cause a decrease in 5α-reduced androgen production, which is probably due to a decrease in 5α-reductase activity. The subsequent increase in oestradiol production corresponds with an increase in aromatase activity. The results indicate that the decrease in 5α-reductase activity as observed in ovaries of spontaneously ovulating rats might be caused by the gradual increase in serum LH, which has been found to occur during the last week before first ovulation.

J. Endocr. (1985) 107, 113–119