Thyroid hormone deiodinases in the central and peripheral nervous system.

Thyroid hormones play a critical role in development and functioning of the nervous system. Deiodinases (type 2 [D2] and type 3 [D3]) contribute to the control of thyroid hormone action in the nervous system by regulating the local concentrations of triiodothyronine (T(3)), the main active thyroid hormone. Most brain T(3) is indeed locally formed by deiodination of thyroxine (T(4)).

Induction of type 2 iodothyronine deiodinase in astrocytes after transient focal cerebral ischemia in the rat.

This study investigated the expression of deiodinases of thyroid hormones in the rat brain after transient occlusion of the middle cerebral artery. The activity of type 2 deiodinase (D2), which catalyzes the deiodination of thyroxine into the more active thyroid hormone 3,5,3'-triiodothyronine, was strongly increased by cerebral ischemia at 6 and 24 hours in the striatum and at 24 hours in the cerebral cortex. The activity of type 3 deiodinase, which catalyzes the inactivation of thyroid hormones, was not affected by ischemia.

The role of MAP kinases in rapid gene induction after lesioning of the rat sciatic nerve.

Lesion of the sciatic nerve caused a rapid activation of p38MAP kinase in the injured nerve adjacent to the site of transection. This activation was detectable 3 min after lesioning, increased during the next 15 min and remained high for several hours. Erk1/2 activation was also observed as early as 15 min after lesioning.

Multidrug resistance genes and p-glycoprotein in the testis of the rat, mouse, Guinea pig, and human.

Study of the multidrug resistance phenomenon in tumor cell lines has led to the discovery of the product of the multidrug resistance (MDR) type 1 genes, the plasma membrane P-glycoprotein (P-gp) that functions as an energy-dependent pump for the efflux of diverse anticancer drugs. P-gp was also recently identified in normal epithelial cells with secretory/excretory functions and in the endothelial cells of the capillary blood vessels in the brain and the testis. These endothelial cells are key elements of the blood-brain and blood-testis barriers, respectively.

Type 2 deiodinase in the peripheral nervous system: induction in the sciatic nerve after injury.

Thyroid hormones are essential for the development and function of the brain and also for the maturation and repair of the peripheral nervous system. In the brain, most of the 3,5,3'-triiodothyronine is locally produced by 5'-deiodination of thyroxine catalyzed by the type 2 deiodinase. The absence of any information about thyroid hormone metabolism in the peripheral nervous system prompted us to study the expression of type 2 deiodinase (mRNA and activity) in the peripheral nervous system.

Induction of type 3 iodothyronine deiodinase by nerve injury in the rat peripheral nervous system.

Thyroid hormones are essential for the development and repair of the peripheral nervous system. The type 2 deiodinase, which is responsible for the activation of T(4) into T(3), is induced in injured sciatic nerve. To obtain information on the type 3 deiodinase (D3) responsible for the degradation of thyroid hormones, we looked for its expression (mRNA and activity) in the sciatic nerve after injury.

Reversible induction of rat hepatoma cell polarity with bile acids.

A dynamic model for inducing and isolating polarized cell colonies from differentiated rat hepatoma was established with chenodeoxycholic acid (CDCA). Cells were treated with 75 microM CDCA in a 1% solvent mix (DMSO/ethanol: 0.5%/0.

Neurosteroid progesterone is up-regulated in the brain of jimpy and shiverer mice.

Concentrations of neurosteroids have been measured in the brains of postnatal myelin mutants jimpy (jp) and shiverer (shi) mice and of their normal controls. Progesterone (PROG) concentrations were increased more than threefold in the brains of mutant mice. Marked astroglial reaction occurs in the brains of jp mice and to a much smaller extent in shi ones.

Stage-specific expression of the immunophilin FKBP59 messenger ribonucleic acid and protein during differentiation of male germ cells in rabbits and rats.

FKBP59 is an immunophilin that binds the immunosuppressant drugs FK506 and rapamycin. It is a 90-kDa heat shock protein (hsp90)-binding protein that was originally discovered as a member of steroid receptor complexes. FKBP59 is ubiquitous and well conserved, and it appears to be a multifunctional protein.

Lead affects steroidogenesis in rat Leydig cells in vivo and in vitro.

Lead is known to impede the male reproductive function, however, the mechanisms through which the adverse effects are mediated are not clearly elucidated. In order to get insight into those mechanisms, we have examined the effects of lead on the biosynthesis of steroid hormones by Leydig cells in the rat. To determine whether lead has a direct action on Leydig cells, we have compared the concentrations of testosterone secreted by Leydig cells in ex vivo experiments after animals had been injected with high doses of lead and in vitro experiments with Leydig cells from normal rats maintained in culture in presence or absence of lead.

Androgen biosynthesis in the stomach: expression of cytochrome P450 17 alpha-hydroxylase/17,20-lyase messenger ribonucleic acid and protein, and metabolism of pregnenolone and progesterone by parietal cells of the rat gastric mucosa.

Dehydroepiandrosterone (DHEA) and its conjugates persist in the rat brain, for up to 1 month after ablation of both adrenals and gonads. Since DHEA synthesis in brain from pregnenolone (PREG) was excluded, we have considered other tissular sources including the digestive tract. In situ hybridization with specific oligonucleotide probes showed that the parietal cells of the gastric mucosa, contrary to other cell types, strongly expressed P450(17) alpha messenger RNA.

Corticotropin-releasing hormone (CRH) is expressed at the implantation sites of early pregnant rat uterus.

We have shown previously that the epithelial cells of human endometrium produce CRH. The biological role of endometrial CRH is not yet known. Among other things, CRH appears to be involved in the inflammatory process, acting as an autocrine/paracrine proinflammatory regulator.

Suppressed expression of the cytochrome P45017 alpha protein in the testicular feminized (Tfm) mouse testes.

The testes of testicular feminized (Tfm) mice synthesize and secrete abnormally low amounts of testosterone, as a consequence of selectively decreased cytochrome P450(17 alpha) activity. To investigate the mechanism of this deficiency, three steroidogenic enzymes were immunolabeled in the testes of normal and Tfm adult (2.5-6 month old) mice.

Astrocytes and neurosteroids: metabolism of pregnenolone and dehydroepiandrosterone. Regulation by cell density.

The rat central nervous system (CNS) has previously been shown to synthesize pregnenolone (PREG) and convert it to progesterone (PROG) and 7 alpha-hydroxy-PREG (7 alpha-OH PREG). Astrocytes, which participate to the regulation of the CNS function, might be involved in the metabolism of neurosteroids. Purified type 1 astrocytes were obtained from fetal rat forebrain with the use of selective culture conditions and were identified by immunostaining with specific antibodies (GFAP+, A2B5-).

Immunoreactive cytochrome P-450(17 alpha) in rat and guinea-pig gonads, adrenal glands and brain.

The cytochrome P-450(17 alpha)-hydroxylase, 17----20 lyase (P-450(17 alpha)) is the key enzyme responsible for the biosynthesis of androgens in steroidogenic organs. Its cellular localization has been examined with an immunohistochemical technique. In immature rat ovary, P-450(17 alpha) was first detected in sparse interstitial cells on postnatal Day 8.

Neurosteroids: biosynthesis, metabolism and function of pregnenolone and dehydroepiandrosterone in the brain.

Pregnenolone (P) and dehydroepiandrosterone (D) accumulate in the brain as unconjugated steroids and their sulfate (S) and fatty acid (L) esters. The microsomal acyl-transferase activity is highest in immature (1-3 weeks old) male rats. The immunocytochemical and biochemical evidence for P biosynthesis by differentiated oligodendrocytes is reviewed.

The cholesterol side-chain cleavage complex in human brain white matter.

Abstract Specific antibodies obtained in rabbits after injection of bovine cholesterol side-chain cleavage enzymes cytochrome P-450scc, adrenodoxin and adrenodoxin-reductase were used for immunohistochemical studies in human brain. The three enzymes were co-localized in the white matter of the cerebellum. This observation strongly suggests the existence of steroidogenic activity in human oligodendrocytes, as previously reported in the rat, and suggests that the concept of 'neurosteroids' can be applied to Delta5-3ss-hydroxysteroid metabolites of cholesterol that accumulate in human brain.

Cell-specific variations and hormonal regulation of immunoreactive cytochrome P-450scc in the rat ovary.

Specific rabbit antibodies to the bovine cholesterol side-chain cleavage cytochrome P-450 (P-450scc) were used to cross-react with the enzyme in the rat ovary. The luteal cells of cyclic, pregnant, and pseudopregnant rats were immunostained. P-450scc was also expressed in the interstitial cells of prepubertal and cyclic adult rats, and in the thecal cells lining the preovulatory follicles.

Neurosteroids: cytochrome P-450scc in rat brain.

The steroid hormones corticosterone and testosterone are supplied to the central nervous system by endocrine glands, the adrenals and gonads. In contrast, the 3 beta-hydroxy-delta 5-derivatives of cholesterol, pregnenolone and dehydroepiandrosterone, accumulate in the rat brain through mechanisms independent of peripheral sources. Immunohistochemical studies have been performed with specific antibodies to bovine adrenal cytochrome P-450scc, which is involved in cholesterol side-chain cleavage and pregnenolone formation.

Alkaline phosphatase activity in the membrane of Xenopus laevis oocytes: effects of steroids, insulin, and inhibitors during meiosis reinitiation.

The mechanism of steroid hormone-induced reinitiation of meiosis in Xenopus laevis oocytes in vitro involves interaction of the hormone with an ooplasma membrane receptor and early changes of enzymatic activities (adenylate cyclase, p48 protein kinase). In full-grown (stage 6) oocytes, we have observed cytochemically, at the ultrastructural level, alkaline phosphatase activity in the ooplasma membrane of microvilli, its decrease by 2 hr of progesterone action, and its complete disappearance at the time of germinal vesicle breakdown (GVBD). Insulin (30 micrograms/ml) also provoked a decrease of phosphatase activity, although it did not promote GVBD under these circumstances.

Testosterone-induced meiotic maturation of Xenopus laevis oocytes: evidence for an early effect in the synergistic action of insulin.

Meiosis reinitiation in oocytes (stage 5-6 of Dumont) isolated free of follicle cells by collagenase treatment from ovarian pieces of Xenopus laevis, was studied in observing the germinal vesicle breakdown (GVBD) provoked by progesterone and testosterone (0.1 nM-1 microM), alone or in association with insulin (30 micrograms/ml). Testosterone, was more active than progesterone to elicit GVBD in vitro, raising the question of the relative roles of both steroids in the physiological maturation process in vivo.

A Membrane Receptor Mechanism for Steroid Hormones Reinitiating Meiosis in Xenopus Laevis Oocytes*: (progesterone/receptor/adenylate cyclase/protein kinase/membrane).

There is a membrane progesterone receptor in Xenopus laevis oocytes that undergo meiosis under steroid exposure. Early responses include a decrease of leucine uptake, a decrease of adenylate cyclase and alkaline phosphatase activities, and a decrease of the phosphorylation of a specific p48 protein in the membrane. These results are compatible with a decrease of membrane fluidity brought about by the hormonal message.

Induction of germinal vesicle breakdown in Xenopus laevis oocytes: synergistic action of progesterone and insulin.

Progesterone is the hormone that reinitiates the meiotic division of amphibian oocytes and insulin and insulin-like growth factors are also active on defolliculated oocytes in vitro. We have studied Xenopus laevis oocytes (stage 5-6) of different hormonal sensitivities, obtained from unstimulated and from human chorionic gonadotrophin-stimulated females. Some oocytes from unstimulated females were also precultured with a subthreshold level of progesterone.

[Potentiation of the effect of steroid hormones (progesterone and testosterone) inducing the reinitiation of meiosis in Xenopus oocytes in vitro, by inhibitors of phosphatase activity].

The induction of meiosis reinitiation by steroid hormones (progesterone and testosterone) in Xenopus laevis oocytes was studied in vitro in presence of inhibitors of phosphatase activity such as beta-glycerophosphate, considered as a competitive inhibitor, and the two ions, Zn++ and MoO--4. Kinetics of the germinal vesicle breakdown indicating the reinitiation of meiosis, have shown that while these phosphatase inhibitors were not active by themselves under the present experimental conditions, they enhanced the process elicited by progesterone or testosterone.

Induction of germinal vesicle breakdown in Xenopus laevis oocytes: response of denuded oocytes to progesterone and insulin.

Denuded oocytes freed of their vitelline envelope have been prepared by two methods, enzymatically with pronase and manually by microdissection. The response of denuded oocytes to progesterone, in terms of germinal vesicle breakdown (GVBD), was similar to that obtained with defolliculated oocytes (separated with collagenase from follicle cells, but still keeping their vitelline membrane). The same conclusion was drawn with respect to morphological features of the oocyte surface observed by transmission and scanning electron microscopy, before and after progesterone-induced GVBD.