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F Vilchis

Publications and source records attributed to F Vilchis.

35 records · Page 2Linked to original sources
PubMed1997

Codon 89 polymorphism of the human 5alpha-steroid reductase type 2 gene.

The existence of a genetic polymorphism within the coding region of the human 5alpha-steroid reductase type 2 (5alpha-SR2) gene is reported in a Mexican population. Genotypic variation was assessed in 100 unrelated, healthy volunteers (50 males; 50 females), using single-stranded conformational polymorphism and direct sequencing analysis. Examination of exon 1 DNAs disclosed the presence of sequences encoding for valine (GTA) or leucine (CTA) at codon 89 of the gene. Of the subjects screened, 45% were homozygous for GTA (89Val), 50% had a heterozygous pattern GTA/CTA (89Val/89Leu) and the remaining 5% were homozygous for CTA (89Leu). These data support the view that the G/C condition at codon 89 of the 5alpha-SR2 gene represents a silent polymorphism which does not alter phenotypical development in the human.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1996

Identification and androgen regulation of a 156-kDa hemeprotein in the harderian gland of the Syrian hamster.

An abundant cytosolic protein from hamster Harderian glands was partially purified by ultracentrifugation and analyzed on SDS-PAGE. This 156-kDa protein occurs in male but not in female glands. Castration of adult males for different time periods resulted in a gradual disappearance of the protein over 5 weeks. Daily administration of testosterone, 5 alpha-dihydrotestosterone, 3 alpha- or 3 beta-androstanediol (0.5 mg/day/15 days), to castrated males maintained the levels of the protein. In females, the protein was induced with the same compounds but not with 17 beta-estradiol. These observations indicate an androgen-mediated hormonal regulation. In developing animals, the 156-kDa protein was detectable in glands of 2-week-old males and females; it was not detected in 4-week-old females. Its specific reactions to 3,3',5,5'-benzidine derivatives strongly suggest heme groups associated with this Harderian gland protein.

Androgens↗
PubMed1995

Molecular interactions of levonorgestrel and its 5 alpha-reduced derivative with androgen receptors in hamster flanking organs.

The 5 alpha-reduction of levonorgestrel (LNG) as well as its binding capacity to the androgen receptors of the hamster flank organ were investigated. Furthermore, the effects of LNG and its 5 alpha-reduced metabolite in the flank organ test and on [U-14C]glucose incorporation into lipids by this tissue were determined. Homogenates from female hamster flank organs were incubated in the presence of [3H]LNG at pH 7.4. The radioactive 5 alpha-LNG metabolite was isolated and its purity was assessed. Competition experiments for androgen binding receptors were carried out with 1.38 nM [3H-7 alpha-17 alpha]dimethyl-19- nortestosterone (DMNT), Kd, plus a range of increasing concentrations of the different unlabeled steroid hormones. The flank organ test was performed in vivo, and [U-14C]glucose incorporation into lipids was determined under organ culture conditions. The 5 alpha-LNG had the same binding capacity to androgen receptors (AR) as LNG in male flank organs. The flank organ test demonstrated that 5 alpha-LNG activity was similar to that observed for levonorgestrel and testosterone (T) on gonadectomized male hamster flank organs. Topical applications of LNG or 5 alpha-LNG increased [U-14C]glucose incorporation into lipids in a way similar to that of T. The overall data indicate that LNG and 5 alpha-LNG produced androgenic activity in the lipid pathway of male flank organs, and that 5 alpha-reduction is not essential for the LNG effect on this tissue.

Animals↗
PubMed1995

Pituitary-ovarian dysfunction in rats with induced nephrotic syndrome.

The reproductive hormonal profile was evaluated in female rats with experimental nephrotic syndrome induced with a single subcutaneous dose of puromycin aminonucleoside (PAN). Serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (P4), testosterone and 17 beta-estradiol (E2) were determined sequentially in control and experimental groups on days 1, 3, 7 and 10 after PAN administration. Prolactin levels were also assessed on day 10. In both groups, vaginal smears were taken daily throughout the study to evaluate cyclic histological changes. At the end of the experiment the histological appearance of the ovaries was evaluated by light microscopy. Nephrotic rats had a rapid loss of the estrous cycle starting on day 4, which set them at diestrus. At the same time the hormonal evaluation indicated a gradual decrease in E2, LH and P4 concentrations, starting from days 3, 7 and 10, respectively. No significant changes were noted in FSH or testosterone values. Besides, on day 10, prolactin concentrations remained unmodified. Even though most hormonal levels were found low on day 10, all values except E2 (undetectable) corresponded to the interval reported for the diestrus phase. Likewise, histological examination of ovarian tissue from nephrotic rats showed a considerable increase in the number of atretic follicles. These findings indicate that female rats with nephrotic syndrome develop an important endocrine dysfunction that probably involves steroidogenic tissues (ovary and/or adrenal gland), and suggest the existence of a hypothalamic-hypophyseal impairment.

Animals↗
PubMed1994

Steroid 5 alpha-reductase activity in the harderian glands of male and female Syrian hamster (Mesocricetus auratus).

The activity of the enzyme steroid 5 alpha-reductase in Harderian glands of adult syrian hamsters was assessed by measuring the conversion of testosterone (T) to 5 alpha-dihydrotestosterone (DHT). The optimal conditions for this reaction were determined in vitro using whole gland homogenates and [3H]testosterone as substrate. Enzyme activity was maximal at pH 5.5. The Michaelis-Menten constant of the Harderian enzyme for T was 4.6 +/- 1.2 x 10(-6) M in females and 4.2 +/- 0.39 x 10(-6) M in males, estimated by Eadie-Hofstee plots. On the basis of relative maximum velocity values, there was 9 or 10 times more 5 alpha-reductase in females (2.8 +/- 0.67 nmol/mg protein/hr) than in males (0.289 +/- 0.029 nmol/mg protein/hr). Consistently, glands of intact male hamsters had lower 5 alpha-reductase activities than those of females. Castration of males significantly increased the enzymatic activity, which within 4 weeks reached female-like values. The levels of 5 alpha-reductase mRNA also increased with castration. There was a direct correlation between activity and mRNA levels of the enzyme in castrated male glands. Further, the administration of T or DHT to ovariectomized hamsters led to intact male values in the enzymatic activity of the gland. The sex differences in 5 alpha-reductase activity may be of relevance to the differential regulation exerted by androgen upon the physiology of male and female glands. The results are consistent with the view that 5 alpha-dihydrotestosterone is the active androgen in the Harderian gland.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1992

Mechanisms of hormonal and antihormonal action of contraceptive progestins at the molecular level.

19-Nor synthetic progestins undergo extensive metabolism at the target cells. The resulting metabolic conversion products interact with putative steroid receptors within the cells, and through those interactions, they may exert either agonistic, synergistic and antagonistic hormonal effects. Studies conducted in our laboratories have disclosed that norethisterone (NET) and D-(1) norgestrel (LNG), two widely used contraceptive progestins, are biotransformed to several A-ring reduced (dihydro and tetrahydro) derivatives. The resulting metabolites 5 alpha-dihydro NET (5 alpha-NET) and 5 alpha-dihydro LNG bind with relative high affinity to the progesterone and androgen receptors. To gain insight into the underlying molecular events mediating the mode of action of NET and its neutral metabolites, we have examined the expression of their biological effects at target organs by using the rabbit uteroglobin gene model and the beta-glucuronidase activity of the mouse kidney. The results of a series of experiments seem to indicate that the enzyme-mediated formation of the 5 alpha (trans A/B ring junction) NET derivative results in a significant diminution of its progestational and androgenic potencies. Furthermore, 5 alpha-NET acquire a potent anti-progestational/contragestational effect as assessed in the female rabbit. These results demonstrated that 5 alpha-reduction of 19-nor progestins exerts a paradoxical effect, at least in terms of their hormone-like effects. The overall data are in line with the concept that metabolism of synthetic progestins at hormone-sensitive organs modulates their mechanisms of action.

Animals↗
PubMed1992

Mechanism of action of levonorgestrel: in vitro metabolism and specific interactions with steroid receptors in target organs.

Levonorgestrel (LNG) is a synthetic steroid that displays potent progestational and androgenic effects but it lacks estrogen-like activity. To examine the mode of action of this progestin, we studied its metabolism in vitro in target organs and the specific interactions of LNG and its metabolites with putative steroid receptors. The results demonstrated that [3H]LNG was efficiently converted to A-ring reduced derivatives when incubated with rat hypothalamus and pituitary. Under optimal incubation conditions, [3H]5 alpha-dihydro LNG (5 alpha-LNG) and [3H]3 alpha,5 alpha-tetrahydro LNG (3 alpha,5 alpha-LNG) were identified as the major metabolic conversion products, while [3H]3 beta,5 alpha-LNG formation occurred to a lesser extent. A-ring reduction of LNG was NADPH-dependent. Assessment of the relative binding affinities of LNG and its derivatives to progesterone (PR), androgen (AR) and estrogen (ER) receptors by displacement analysis revealed that unchanged LNG binds with high affinity to PR and AR but not to ER. 5 alpha-LNG exhibited a diminished though significant interaction with PR and an enhanced binding affinity for AR as compared with LNG, indicating that 5 alpha-reduction of LNG increases its affinity for AR. The most striking finding was that further reduction of the 5 alpha-LNG molecule at C-3 abolished its binding activity to PR, AR, and even to ER. The overall data provides a plausible explanation for the lack of estrogen agonistic action of LNG and for its potent progestational and androgenic effects.

Animals↗
PubMed1991

Steroid hormone binding in the Harderian gland of birds: characteristics of the androgen, estrogen, and progestin receptors of Anas platyrhynchos and Gallus domesticus.

A series of experiments was carried out in immature female chicks and ducks to establish whether the avian Harderian gland contains specific receptors for sex steroids. Cytosol preparations of Harderian glands were submitted to hormone saturation analysis using radiolabeled estradiol, ORG-2058, and dimethylnortestosterone as ligands. In addition, the sedimentation characteristics of the hormone-receptor complexes were studied by ultracentrifugation of linear sucrose gradients. The presence of high affinity binding sites for estrogens (Kd = 2.4 and 1.6 nM), progestins (0.8 and 1.0 nM), and androgens (1.0 and 1.0 nM) was indicated in the chick and duck glands, respectively. The sedimentation coefficients were 7-7.5 S, 7-8 S, and 8 S for estrogen, progestin, and androgen receptor-ligand complexes, respectively. The concentration of the androgen receptor was significantly higher in chick than in duck Harderian glands whereas the estrogen and progestin receptor concentrations were similar in both species. A striking finding was the presence of progestin receptors, which apparently do not exist in the glands of many mammals. Priming with estrogens did not modify the concentration of ORG-2058 binding sites in either species studied, indicating that gland progestin receptor is not estrogen-regulated. Overall the data suggest intracellular mechanisms whereby circulating gonadal hormones regulate avian Harderian gland function.

Animals↗
PubMed1989

Steroid hormone receptors and the sexual phenotype of the Harderian gland in hamsters.

To investigate the participation of intracellular steroid hormone receptors in the sexual transformation process of the Harderian gland, a series of experiments were undertaken in adult golden hamsters. The invitro labelling of cytosolic steroid-binding sites with appropriate radioligands revealed the presence of androgen, oestrogen and glucocorticoid but not progestin receptors in the glands from animals of both sexes. The androgen receptor of the female gland was further characterized because it was found to be the predominant intracellular steroid receptor. Studies of binding kinetics using [3H]7 alpha,17 alpha-dimethyl-17 beta-hydroxy-4-oestren-3-one (DMNT) as ligand, demonstrated a high affinity androgen-binding site with an apparent dissociation constant (Kd) of 0.7 nmol/l and maximal saturation binding capacity of 84.0 +/- 3.0 (S.D.) fmol/mg protein. Specificity of the androgen receptor was assessed by displacement analysis; DMNT, 5 alpha-dihydrotestosterone, testosterone and 3 alpha-androstanediol were efficient competitors for the androgen-binding site, while oestradiol-17 beta, progesterone and dexamethasone exhibited very little, if any, competitive potency. The sedimentation coefficient of the androgen receptor in sucrose density gradients was 8-9 S. These data indicate that the physicochemical characteristics of the androgen receptor from the female gland are similar to those previously described in the male gland. The striking observation of a complete lack of oestrogen-inducible and oestrogen-insensitive progestin receptors in glands cytosol, even after stimulation with cholera toxin, adds further support to the concept that the androgen receptor is the key molecule mediating the hormone-induced sexual transformation of the Harderian gland in this species.

Animals↗
PubMed1987

The metabolism of 19-nor contraceptive progestins modulates their biological activity at the neuroendocrine level.

In this communication, a series of studies from our laboratory dealing with the mechanism of action of 17 alpha-ethinyl derivatives of 19-nor testosterone are reviewed. The administration of norethisterone (NET) to long-term castrated female rats induces the nuclear translocation of pituitary estradiol receptors and is followed by some estrogenic-like effects at the hypothalamic-pituitary unit. It is established that an A-ring reduced metabolite of NET, the 3 beta,5 alpha-tetrahydro NET derivative, is responsible for the observed in vivo estrogenic effects of the parent compound. 3 beta,5 alpha-NET binds to the estrogen receptor and is efficient in inducing the pituitary estrogen-dependent progesterone receptor and in increasing the uterine weight in long-term castrated rats. Furthermore, administration of 3 beta,5 alpha-NET and the 5 alpha-reduced metabolite of NET (5 alpha-NET) are able to inhibit the release of gonadotropins in the castrated animal to a greater extent than NET. Moreover, pretreatment with tamoxifen, an estrogen binding site competitor, results in a significant diminution of the antigonadotropic potency of 3 beta,5 alpha-NET but not of the 5 alpha-NET, which is only inhibited by the administration of cyproterone acetate. These findings underline the importance of the metabolic rate of NET for the expression of its biological effects at the hypothalamic-pituitary unit.

Animals↗
PubMed1987

Hormone regulation of the rodent Harderian gland: binding properties of the androgen receptor in the male golden hamster.

Studies were conducted in castrated golden hamsters to assess whether sexual dimorphism and sensitivity to sex steroid hormones in the rodent Harderian gland are mediated by an interaction of androgens with specific intracellular receptors. Physical properties, binding kinetics and stereospecificity of the androgen receptor were analysed using [3H]mibolerone as the radioligand. The presence of [3H]mibolerone-androgen receptor complexes with a sedimentation coefficient of 7-8S was demonstrated in Harderian gland cytosol by a linear sucrose gradient ultracentrifugation technique using a vertical rotor. Kinetic analysis revealed an androgen-binding site with an apparent dissociation constant of 0.3 +/- 0.07 (S.D.) nmol/l and a saturation binding capacity of 113 +/- 15 fmol/mg protein. Displacement studies indicated that unlabelled mibolerone, methyltrienolone, 5 alpha-dihydrotestosterone and testosterone were efficient competitors for the androgen-binding sites, while progesterone, 17 beta-oestradiol, dexamethasone, dehydroepiandrosterone, ethiocholanolone and 5 alpha-16-androsten-3-one were not. Experiments in longterm castrated animals revealed that the Harderian gland androgen receptor concentration and sedimentation coefficient remained unmodified. The results of these studies were interpreted as demonstrating the presence of a specific high-affinity intracellular androgen receptor in the male hamster Harderian gland.

Animals↗
PubMed1986

Evidence that a non-aromatizable metabolite of norethisterone induces estrogen-dependent pituitary progestin receptors.

Neutral reduced metabolites of norethisterone (NET) specifically interact with intracellular estrogen receptors in target organs. To determine if this interaction can effectively initiate estrogen-dependent cellular responses, the effects of an A-ring-reduced NET derivative upon the induction of cytosol-located pituitary progestin receptors (PR) and uterine growth were studied in adult castrated female rats. Different doses of 17 alpha-ethynyl-5 alpha-estran-3 beta, 17 beta-diol (3 beta, 5 alpha-NET) were s.c. administered to ovariectomized animals for 6 days. 17 beta-Estradiol (E2) and oil-treated rats served as experimental controls. Pituitary PR were labeled in vitro by a post-gradient technique using [3H]ORG-2058 as the ligand. PR binding specificity was determined by the use of an excess of radioinert steroids. The results demonstrated that administration of 3 beta, 5 alpha-NET induced specific 8-9S pituitary cytosol PR in a dose-dependent manner. Binding properties of the 3 beta, 5 alpha-NET-induced progestin binding sites (Kd = 1.0 X 10(-9) M; NBS = 1.2 X 10(-9) M) appear indistinguishable from those induced by E2. In addition, 3 beta, 5 alpha-NET administration resulted in a significant increase in uterine weight at the expense of myometrium and endometrium growth in a similar fashion to that observed in the E2-treated group. When 3 alpha, 5 alpha-epimeric alcohol (3 alpha, 5 alpha-NET) was administered, induction of pituitary PR and uterine growth were also observed although to a lesser extent. Inasmuch as the results demonstrate that neutral non-aromatizable NET metabolites induce biochemical and morphological estrogenic responses, they offer an alternative explanation for the mechanism of estrogen-like action of this synthetic contraceptive progestin.

Animals↗
PubMed1986

A-ring reduction enhances the antigonadotropic potency of norethisterone.

To assess whether structural modifications on the A-ring of norethisterone (NET) could modify its antigonadotropic potency, comparative studies using NET, 5 alpha-dihydro NET (5 alpha-NET) and its 3 beta,5 alpha and 3 alpha,5 alpha tetrahydro derivatives in castrated adult rats were undertaken. The antigonadotropic effect of these compounds was evaluated by measuring the serum and pituitary immunoreactive concentrations of LH and FSH following their chronic sc administration to animals depleted of progesterone receptors. The results demonstrated that 3 beta,5 alpha-NET and 5 alpha-dihydro-NET exhibited a significantly greater gonadotropic inhibiting activity as compared with that of their parent compound. The simultaneous administration of tamoxifen with 3 beta,5 alpha-NET resulted in a significant diminution of its antigonadotropic potency, particularly for LH. These data indicate that the potent antigonadotropic effect of 3 beta,5 alpha-NET metabolite was mediated via oestrogen receptors. The LH inhibitory activity of 5 alpha-dihydro-NET was not suppressed by the non-steroidal antioestrogen administration, thus suggesting that 5 alpha-NET might exert its effect via androgen receptors. The overall data were interpreted as demonstrating that metabolic conversion products of NET exhibit potent antigonadotropic effect. The data are consistent with an A-ring enhancement of the antigonadotropic potency of this synthetic progestin and open an alternate approach to the development of fertility regulating agents.

Animals↗
PubMed1985

Stereospecificity of the intracellular binding of norethisterone and its A-ring reduced metabolites.

The interaction of norethisterone (NET) and four A-ring reduced metabolites of NET with cytosol receptors for progesterone (PR), androgen (AR), and estrogen (ER) was investigated. Cytosol preparations from: uteri of adult estrogen-primed castrated rats, ventral prostates of adult castrated rats and uteri of immature rats were used as the source of PR, AR, and ER respectively. 3H-Labeled ORG-2058, R-1881, and 17 beta-estradiol were used as the radioligands. The results of competitive studies disclosed that: the most efficient competitor for PR binding sites was NET (Ki = 1.1 X 10(-7) M) followed by 5 alpha-dihydro NET (5 alpha-NET), whereas the 3 alpha,5 alpha; 3 beta,5 alpha and 3 alpha,5 beta-tetrahydro NET derivatives were ineffective the most efficient competitor for AR binding sites was 5 alpha-NET (Ki = 1 X 10(-8), immediately followed by NET, while the three tetrahydro NET derivatives were not competitors and remarkable competition for ER binding sites was only exhibited by the 3 beta,5 alpha-tetrahydro NET derivative (Ki = 4.6 X 10(-8) M) and to a lesser extent by its 3 alpha,5 alpha-epimeric alcohol, while NET and 5 alpha-NET were completely ineffective. These findings demonstrate the stereospecificity of the intracellular binding of NET and its reduced metabolites with cytosol steroid putative receptors, and provide biochemical support to the understanding of the variety of hormone-like effects observed after the in vivo administration of NET.

Animals↗
PubMed1983

Interaction of medroxyprogesterone acetate with cytosol androgen receptors in the rat hypothalamus and pituitary.

The binding of medroxyprogesterone acetate (MPA) with cytosol androgen receptors from rat pituitary and hypothalamus was studied. The pituitary and hypothalamic cytosol androgen receptors from adult castrated female rats were in vitro labeled using 3H natural (testosterone (T) and 5 alpha-dihydrotestosterone (DHT] and [3H]synthetic (methyltrienolone) androgens as radioligands. The [3H]androgen-receptor complexes sedimented with a coefficient of 8S in linear sucrose gradients. When incubated with an excess of radioinert MPA, specific binding was abolished indicating interaction of MPA with androgen receptors. Furthermore specific [3H]MPA-androgen cytosol receptor complexes could be identified in these neuroendocrine tissues when a post-gradient receptor labeling technique was used in the absence or presence of radioinert MPA, DHT, and triamcinolone acetonide. A study of binding kinetics disclosed that the equilibrium dissociation constant and saturation binding capacity for the MPA binder, were similar to those exhibited by DHT binding to androgen receptors in both studied tissues under identical experimental conditions. The overall results were interpreted as demonstrating that MPA interacts with cytosol steroid receptors other than those of progesterone in the rat hypothalamus and anterior pituitary. The data are consistent with MPA binding to androgen receptors.

Animals↗