Subcellular Distribution of 3α-Hydroxysteroid Dehydrogenase and Antiestrogen Action on Androgen-Metabolizing Enzymes in Rat Pituitary Gland

Abstract: Gonadectomy of male rats led to a threefold increase of 3α-hydroxysteroid dehydrogenase (3α-HSDH) activity in pituitary homogenates that could be completely reversed by chronic administration of estradiol or 5α-dihydrotestosterone (DHT). 3α-HSDH was found to be distributed mainly between the 10,000 g and 100,000 g sediments from whole homogenates. The microsomal enzyme activity showed a substantial specificity for NADH whereas the cytosolic enzyme (100,000 g supernatant) demonstrated a slight preference for NADPH. The changes in V _max found in homogenates following gonadectomy and gonadal steroid administration reflected changes in NADH- linked activity of the microsomal, but not the cytosolic enzyme. Estradiol-induced suppression of NADH-linked 3α-HSDH activity in pituitary homogenates from gonadectomized rats of either sex was accompanied by a similar suppression of NADPH-linked 5α-reductase activity and a marked decrease of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release. In the ovariectomized rat chronic administration of nonsteroidal antiestrogens had strong estrogenic effects on 3α-HSDH activity and LH release, but not on 5α-reductase activity and FSH release. In the gonadectomized male rat, which was much less sensitive to intrinsic estrogenicity of the antiestrogens tested, nafoxidine completely blocked estradiol-induced suppression of 5α-reductase activity and FSH release and partially antagonized suppression of LH release. The trans -isomeric, substituted triphenylethylenes, tamoxifen, and enclomiphene, as well as nitromifene (mixture of trans and cis isomers) were able partially to counteract estradiol-induced suppression of 5α-reductase, but not 3α-HSDH activity. It is concluded that estradiol action on pituitary 5α-reductase, but not 3α-HSDH activity, involves an estrogen receptor mechanism.     

Steroid hormone effects on growth and apical dominance of sunflower

External application of estradiol-17β increased shoot growth but decreased root growth of sunflower seedlings. It completely inhibited cotyledonary axillary bud development in decapitated plants at the concentration of 1 μg/plant. Concentrations lower than this promoted cotyledonary axillary bud formation. Testosterone on the other hand inhibited both shoot and root growth and promoted cotyledonary axillary bud formation at all the concentrations used. Progesterone at high (0.25,μg/plant) concentration promoted shoot growth but inhibited root growth. A low concentration (0.1 μg/plant) of progesterone produced the opposite effect.     

Failure to detect delta 5-3 beta-hydroxysteroid oxidoreductase activity in the preimplantation rabbit embryo

Preimplantation rabbit embryos were incubated with pregnenolone and dehydroisoandrosterone under conditions which gave formazan precipitation by the histochemical technique. The metabolic fate of the labeled steroids was assessed simultaneously. There was no concomitant transformation of pregnenolone to progesterone and dehydroisoandrosterone was not transformed to androstenedione. It is concluded that the formazan precipitation is coupled with an "activity" other than delta 5-3 beta-hydroxysteroid oxidoreductase.     

Stabilization of rat liver lysosomes by new anti-inflammatory steroids in vitro

Steroid acid esters, synthesized by modifying the 17-ketol side chain of prednisolone, were tested for their in vitro ability to stabilize heavy mitochondrial lysosomes prepared from rat liver. Membrane stabilization was determined by assessing capability of steroids to decrease extrusion of the marker enzymes (acid phosphatase, beta-glucuronidase and aryl sulfatase) from lysosomes incubated in hypo-osmotic sucrose-Tris acetate buffer. Results indicated that prednisolone (1) significantly inhibited the lysosomal release of acid phosphatase as did the new anti-inflammatory steroid, methyl 20-dihydroprednisolonate. Methyl prednisolonate exhibited weak membrane stabilization capacities and 20-dihydroprednisolonic acid, a metabolic product of methyl 20-dihydroprednisolonate, showed virtually no membrane stabilization.     

Androgen metabolism in male and female breast tissue

Incubation studies have been carried out using normal breast tissue and breast tissue from patients with gynecomastia, mammary dysplasia and breast carcinoma to determine the pattern of androstenedione metabolism. All tissues formed estrone (E₁) and testosterone (T) in all incubations. Estradiol (E₂) was isolated in incubations of tissue from 1 to 6 patients with mammary dysplasia, 5 of 6 patients with gynecomastia and in all incubations with normal and carcinoma tissue. Estrone formation was lowest in mammary dysplasia and gynecomastia, and higher in apparently normal breast tissue. The greatest E₁ formation was found in incubations with breast carcinoma tissue, although there was considerable variation within this tissue group. Estradiol formation was low in all tissues, with the highest conversion rates in carcinoma tissue. Testosterone formation in carcinoma tissue was greater than in mammary dysplasia or gynecomastia, but similar to apparently normal tissue. These results indicate that breast tissue from different pathological states varies in its capacity to aromatize androstenedione (A) to estrogenic products and to convert it to other androgens. They have also shown that the pattern of metabolism is distinctive for the nature of the pathological abnormality.     

Phospholipid chain immobilization and steriod rotational immobilization in acetylcholine receptor-rich membranes from torpedo marmorata

1. The ESR spectra of both phosphatidylcholine and phosphatidylethanolamine spin labels reveal an immobilized lipid component (τ_R ? 50 ns), in addition to a fluid component (τ_R ～ 1 ns), in acetylcholine receptorrich membranes prepared from Torpedo marmorata electroplax according to the method of Cohen et al. (Cohen, J.B., Weber, M., Huchet, M. and Changeux, J.P. (1972) FEBS Lett. 26, 43–47). 2. The ESR spectra of the androstanol spin label display a component corresponding to molecules which are immobilized with respect to rotation about the long molecular axis (

), in addition to the fluid lipid bilayer component in which the molecules are rotating rapidly about their long axes (

). This immobilized component is observed throughout the temperature range 2–22°C, at an approximately constant relative intensity of approx. 45% of the total, which is quantitatively the same as previously observed with fatty acid spin labels.     

Biosynthesis of chondroitin sulfate: Microsomal proteoglycans

A microsomal preparation from chick embryo epiphyseal cartilage was incubated with UDP-[¹⁴C]glucuronic acid and UDP-N-acetylgalactosamine to form [¹⁴C] chondroitin-labeled proteoglycan. Two [¹⁴C]proteoglycan populations were obtained which differed in size, [¹⁴C]glycosaminoglycan content, and susceptibility to alkali. One population of [¹⁴C]proteoglycan appeared near the void volume on Sepharose 2B, while the other population was smaller, similar in size to monomer proteoglycan. The larger [¹⁴C]proteoglycan contained long [¹⁴C]chondroitin chains added to short primers; these chains were in part resistant to alkali cleavage from protein. The smaller [¹⁴C]proteoglycan contained mainly [¹⁴C]chondroitin chains of intermediate length added to endogenous chondroitin sulfate; these chains were all susceptible to alkali cleavage from protein. The larger [¹⁴C]proteoglycan may represent a precursor proteoglycan present at the site of glycosaminoglycan chain synthesis.     

Assignment of the gene for glyoxylase I to mouse chromosome 17 by somatic cell genetics

Evidence is presented for the assignment of the gene for glyoxylase I to mouse chromosome 17 using mouse × Chinese hamster somatic cell hybrids. GLO I was not expressed concordantly with any known marker enzymes which represented 11 linkage groups. The presence of chromosome 17 and expression of GLO I were concordant in 31/31 clones. GLO I is thus linked to the H-2 histocompatibility locus in the mouse.     

In vivo metabolism of 3H-testosterone in adult male rats: effects of estrogen administration.

The metabolism of testosterone (T) was studied in normal adult male rats using a constant infusion of trace amounts of the 3H-steroid into a tail vein for 3 h in order to attain a state of equilibrium. Samples of plasma, liver, kidney, prostate, seminal vesicles and muscle were analysed for 3H-testosterone, 3H-5alpha-dihydrotestosterone (5alphaDHT) and 3H-5alpha-androstanediol (Adiol). When compared to the 3H-T level in plasma there were high levels of 3H-T in kidney and of 3H-5alphaDHT in prostate and seminal vesicles. Intraperitoneal estradiol valerate administration (100 mug/day) for 4 days decreased and 3H-5alphaDHT levels in the prostate and seminal vesicles. The estrogen administration increased the T metabolic clearance rate from 17.5 1/24 h/100 g body wt to 22.6 1/24 h/100 g body wt.