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Biomedical subjects

B Eckstein

Publications and source records attributed to B Eckstein.

At least 37 records · Page 2Linked to original sources

Enzymes of steroid hormone metabolism in ovarian tissue of teleosts.

1. Substrate-velocity kinetics and some other characteristics of three enzymes involved in steroid hormone metabolism in the ovary of the common carp, Cyprinus carpio and of the grey mullet, Mugil cephalus were determined. 2. Labeled steroid products of the enzyme reactions were identified by thin-layer and gas-liquid chromatographies, as well as by formation of derivatives. 3. 5 alpha-Steroid reductase of carp ovary was found to have a Michaelis-Menten constant undistinguishable from that of rat ovarian tissue, while the Km of the enzyme in mullet ovarian tissue was found to be different. 4. The Km of 20 alpha-hydroxysteroid dehydrogenase of mullet ovarian tissue was similar to the value reported for mammalian gonadal tissue, but the constant for 17 beta-hydroxy-steroid dehydrogenase in the carp ovary was different. 5. It is for the first time that these enzymes of steroid hormone metabolism have been quantitatively determined in teleost ovarian tissue.

17-Hydroxysteroid Dehydrogenases↗

Effect of human chorionic gonadotropin and prolactin on 20 alpha-hydroxysteroid dehydrogenase activity in granulosa cells of immature rat ovary.

The effects of FSH, hCG, and PRL on the activity of 20 alpha-hydroxysteroid dehydrogenase (20 alpha-SDH) of separate ovarian components of hypophysectomized, diethylstilbestrol-treated rats were studied. Enzyme activity was found to reside mainly in granulosa cells. FSH induced an increase in enzyme activity. A preparation of FSH was purified by adsorbing its LH contamination on rat corpora lutea membranes and by further neutralizing LH traces with an antiserum to the beta-subunit of LH. This purified FSH retained the ability to induce a 6-fold increase in specific enzyme activity in granulosa cells of hypophysectomized, diethylstilbestrol-treated rats. Administration of hCG to rats treated with purified FSH, further enhanced 20 alpha-SDH activity in granulosa cells up to 11.5-fold above control. PRL, which is known to inhibit 20 alpha-SDH activity in regressing rat corpora lutea, suppressed the FSH-induced increase in enzyme activity in the granulosa cells.

20-Hydroxysteroid Dehydrogenases↗

Properties of microsomal delta4-3-ketosteroid 5alpha-reductase in immature rat ovary. Inhibition by estradiol-17beta.

The subcellular distribution of immature rat ovarian 5alpha-reductase has been studied by utilizing 20alpha-hydroxypregn-4-en-3-one as substrate. The main enzyme activity was found to be associated with the microsomal fraction, with lower activities in the 1000 X g and 10 000 X g fractions. The enzyme activity associated with the microsomes exhibited an apparent Km of 3.0 +/- 1.1 micrometer for 20alpha-hydroxypregn-4-en-3-one and 36.3 +/- 6.7 micrometer for testosterone as substrate. Progesterone and testosterone competitively inhibited the 5alpha-reduction of 20alpha-hydroxypregn-4-en-3-one; estradiol-17beta was found to be a noncompetitive inhibitor of this reaction. A concentration of estradiol-17beta as low as 1 micrometer when added to 25 micrometer concentration of 20alpha-hydroxypregn-4-en-3-one exerted a significant inhibition of 5alpha-reductase activity.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Changes in steroid concentration in the ovaries of immature rats treated with pregnant mare serum gonadotrophin and human chorionic gonadotrophin.

The concentrations of testosterone, progesterone and 20alpha-hydroxypregn-4-en-3-one (20alpha-OHP) were measured in the ovaries of immature rats in which ovulation was induced by treatment with pregnant mare serum gonadotrophin (PMSG) and, 48 h later, with human chorionic gonadotrophin (HCG). The concentration of testosterone in the tissue increased significantly 48 h after treatment with PMSG, reached a peak 4 h after the administration of HCG and declined to the basal level 4 h later. Increases in the levels of progesterone and 20alpha-OHP were observed 4 h after the administration of HCG. Whereas the level of 20alpha-OHP continued to rise during the subsequent 30 h, progesterone levels declined near the presumed time of ovulation (12 h after administration of HCG). It is concluded that 20alpha-hydroxysteroid dehydrogenase activity is present in the immature rat ovary before ovulation and that an increase in the production of testosterone in the ovaries of rats treated with PMSG and HCG precedes increased production of progesterone and 20alpha-OHP in these ovaries.

20-alpha-Dihydroprogesterone↗

Identification of two 5alpha-reduced pregnanes as major metabolites of progesterone in immature rat ovaries (1000 x g supernatant) in vitro.

Incubation of the 1000 x g supernatant obtained from 23-day-old rat ovarian homogenate with labeled progesterone resulted in the production of 3 major metabolites; 5alpha-androstane-3alpha,17beta-diol, and two 5alpha-reduced pregnanes that were identified as 3alpha-hydroxy-5alpha-pregnan-20-one and 3alpha,17alpha-dihydroxy-5alpha-pregnan-20-one. The 3alpha,17alpha-dihydroxy-5alpha-pregnan-20-one has not been hitherto isolated from mammalian ovaries. The steroids were identified by their mobilities on thin-layer and gas-liquid chromatography, by mass spectroscopy, derivative formation and by recrystallization to constant specific activity. In another experiment, incubation of the 1000 x g supernatant from 23-day-old rat ovaries with 3alpha-hydroxy-5alpha-pregnan-20-one as substrate resulted in the production of 5alpha-androstane-3alpha,17beta-diol. It is suggested that 5alpha-androstane-3alpha,17beta-diol is produced in immature rat ovaries by a pathway in which the identified 5alpha-reduced pregnanes serve as intermediates.

Androstane-3,17-diol↗

Suppression of luteinizing hormone release by 5alpha-androstane-3alpha, 17 beta-diol and its 3 beta epimer in immature ovariectomized rats.

The effect of 5alpha-androstane-3alpha, 17 beta-diol, its 3 beta epimer and oestradiol benzoate on suppression of LH release after ovariectomy was studied in immature rats. At doses of 50 and 100 mug/100 g body weight/day the 3alpha compound suppressed LH release after ovariectomy to the same extent as 0-1 mug oestradiol benzoate/100 g/day. 3alpha-Androstanediol at a dose of 25 mug/100 g/day suppressed LH release only up to day 45 of life, while the same dose of the 3beta epimer had no effect on LH suppression. The effect of 3beta-androstanediol on inducing precocious vaginal opening was found to be mediated by the ovaries, since it was eliminated by ovariectomy. These results confirm our previous findings on the participation of androstanediol in the normal regulation of LH and in the mechanism of onset of puberty in the rat.

Androstane-3,17-diol↗

Pre-ovulatory changes in steroidogenesis in ovaries from immature rats treated with pregnant mare serum gonadotrophin.

The metabolism of progesterone in the 1000 g supernatant fraction of homogenates of ovaries from PMSG-treated immature rats was determined. As early as 52 h after a single injection of 50 i.u. PMSG, still before the LH surge, 5alpha-pregnane-3alpha, 20alpha-diol was identified as the main metabolite, together with small quantities of 3alpha-hydroxy-kalpha-pregnan-20-one and 5alpha-androstane-3alpha, 17beta-diol. Similar incubations of untreated rat ovaries at the same age did not produce 5alpha-pregnane-3alpha, 20alpha-diol. The quantities of 3alpha-hydroxy-5alpha-pregnan-20-one and 5alpha-androstane-3alpha, 1mbeta-diol were reduced in PMSG-treated rat ovaries as compared with control ovaries. When progesterone metabolism was examined 64 h after PMSG administration, 5-7 h after the peak of LH surge but still before ovulation, 75% of the substrate was converted to 5alpha-pregnane-3alpha, 20alpha-diol, while 3alpha-hydroxy-5alpha-pregnan-20-one as well as 5alpha-androstane-3alpha, 17beta-diol could not be detected.

Age Factors↗

Androstanediol sulphates in peripheral blood of immature rats and some of their biological effects.

The conjugation of 5alpha-androstane-3alpha,17beta-diol (3alpha-A) and its 3beta epimer (3beta-A) was determined in the peripheral blood of immature female rats. About two thirds of these steroids were present in blood as sulphates and one third as glucuronides; no free steroids were detected. Administration of 3beta-A sulphate (25mug/100 g body weight/day) and of 3alpha-A sulphate (50 mug/100 g/day) from day 21 of life until the day of vaginal opening, advanced the day of the first ovulation. Administration of the 3beta-A sulphate did not induce precocious vaginal opening whereas the free alcohol was active in this respect. Implantation of 3beta-A sulphate, but not of the 3alpha epimer, into the basal medial hypothalamus resulted in the dealth of all animals within 24 h.

Androstane-3,17-diol↗