11 beta-Hydroxysteroid dehydrogenase in the rat ovary: high expression in the oocyte.

The enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) catalyses the conversion of physiological glucocorticoids to inactive products, thus modifying the access of glucocorticoids to glucocorticoid and mineralocorticoid receptors. Glucocorticoids may affect ovarian function both indirectly and via binding to ovarian receptors. We have demonstrated 11 beta-HSD bioactivity and mRNA expression in rat ovary in vitro. The enzyme was localized to oocytes and luteal bodies immunohistochemically using two antibodies raised against purified rat liver 11 beta-HSD. These data are supported by in-situ hybridization studies, which also localized 11 beta-HSD mRNA expression to oocytes and luteal bodies. The results suggest that 11 beta-HSD may modulate the effects of glucocorticoid on ovarian function.

11 beta-hydroxysteroid dehydrogenase in vascular smooth muscle and heart: implications for cardiovascular responses to glucocorticoids.

The enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta-OHSD) converts the active glucocorticoid corticosterone to inactive 11-dehydrocorticosterone in the rat (or cortisol to cortisone in man), thereby protecting renal mineralocorticoid receptors from corticosterone or cortisol and allowing preferential access for aldosterone. We have previously demonstrated that cortisol-induced cutaneous vasoconstriction in man is potentiated by the 11 beta-OHSD inhibitor glycyrrhetinic acid, suggesting that 11 beta-OHSD may protect vascular corticosteroid receptors. In this study we report quantitation of 11 beta-OHSD bioactivity in homogenates of rat aorta, mesenteric artery, caudal artery, and heart, expressed as the percent in vitro conversion of 3H-corticosterone to 3H-11-dehydrocorticosterone. Nicotinamide adenine dinucleotide phosphate (NADP+)-dependent 11 beta-OHSD activity was found in all of these tissues and was significantly higher in resistance vessels than aorta (P less than 0.05) [without NADP+: caudal artery (4.2 +/- 0.2%) greater than mesenteric artery (2.5 +/- 0.7%) = heart (1.67 +/- 0.2%) greater than aorta (0.79 +/- 0.2%); with 200 microM NADP+: caudal artery (43.9 +/- 2.1%) greater than heart (20.6 +/- 1.0%) = mesenteric artery (17.7 +/- 3.1%) = aorta (11.4 +/- 0.4%); heart greater than aorta]. All of these were lower than renal cortex (29.4 +/- 1.8% without NADP+; 82.4 +/- 0.4% with NADP+; P less than 0.001). 3H-11-dehydrocorticosterone was the major metabolite of 3H-corticosterone (greater than 97% of 3H-corticosterone metabolized). Reduction of 3H-11-dehydrocorticosterone to 3H-corticosterone was not detected in these experiments. We also report localization of 11 beta-OHSD-like immunoreactivity by immunohistochemistry using antisera raised against rat liver 11 beta-OHSD, and of 11 beta-OHSD messenger RNA expression by in situ hybridization using complementary RNA probes transcribed from complementary DNA encoding rat liver 11 beta-OHSD. We found 11 beta-OHSD immunoreactivity and messenger RNA expression in vascular and cardiac smooth muscle cytoplasm but not in endothelium. Thus, 11 beta-OHSD is appropriately sited to modulate access of corticosterone to vascular receptors and could influence vascular resistance, cardiac output and thereby blood pressure.

11Beta-Hydroxysteroid Dehydrogenase Messenger Ribonucleic Acid Expression, Bioactivity and Immunoreactivity in Rat Cerebellum.

Abstract 11beta-Hydroxysteroid dehydrogenase (11beta-OHSD) metabolizes corticosterone to inactive 11-dehydrocorticosterone and thus protects non-specific mineralocorticoid receptors from exposure to corticosterone in the kidney in vivo. Clearly, 11beta-OHSD might also regulate corticosterone access to glucocorticoid receptors. We have investigated cerebellum, a tissue with high glucocorticoid receptor, but very low mineralocorticoid receptor levels and have shown marked 11beta-OHSD bioactivity with similar co-substrate requirements and inhibition kinetics to the renal enzyme. 11beta-OHSD messenger ribonucleic acid was expressed in cerebellum and was localized in Purkinje and granule cells. This distribution was confirmed immunohistochemically. Thus, we provide evidence for 11beta-OHSD in cerebellum and suggest that it may regulate the access of corticosterone to glucocorticoid receptors in addition to mineralocorticoid receptors.

Bidirectional responsiveness of the pituitary-adrenal system in old and young male and female rats.

Bidirectional responsiveness of the pituitary-adrenal system was examined in young adult (3-5 months) and old (24-26 months) male and female rats. In Experiment 1, an approach-avoidance conflict was created by exposing food- and water-deprived subjects to a flavored solution which had previously been paired with lithium chloride-induced illness in a conditioned taste aversion paradigm. Young and old males and young females elevated plasma corticosterone in response to deprivation; males showed a further elevation when reexposed to the solution, whereas young females did not. Old females did not exhibit a corticosterone elevation to deprivation or upon reexposure. In Experiment 2, water was restricted to a fixed period in the morning when corticosterone levels are typically low. Plasma corticosterone was measured before and after watering. Young and old males and young females reentrained their circadian corticosterone rhythm so that levels were elevated just prior to watering; consumption was followed by a drop in corticosterone. Old females failed to reentrain or to suppress corticosterone secretion upon drinking. These findings indicate that the lability of the pituitary-adrenal system is more markedly affected by senescence in females than in males.

The pituitary-adrenal response to novel stimulation and ether stress in young adult and aged rats.

Pituitary-adrenal responsiveness to stress induced by either psychological (novelty) or physiological (ether) stimuli was examined in young (3 to 6 months) and old (24 to 27 months) male and female rats. In Experiment 1, subjects were placed in a novel environment for 3 min. Blood samples were collected at 0, 15, 30, 45, and 60 min. In Experiment 2, a blood sample was collected from experimental subjects immediately following 15 min in a novel environment; a basal sample was collected from control subjects. In Experiment 3, basal and stress blood samples were collected from subjects exposed to ether vapors. Blood samples were assayed for plasma corticosterone content. The major findings were: (1) no change with age in either sex in basal corticosterone levels or in the time course of the adrenocortical response; (2) no change with age in males in stress-induced increments in corticoids; (3) no change with age in females in corticoid elevations induced by a mild stressor (Experiment 1), but a decrement in elevations induced by more potent stressors (Experiments 2 and 3).

Adjunctive drinking and the pituitary-adrenal response: effects of prior aversive stimulation (preshock).

Two groups of rats were exposed to an intermittent food schedule which induced polydipsia. One group of rats had previously been exposed to inescapable shock. This group showed reduced amounts of polydipsia, and the polydipsia was ineffective in reducing arousal levels (relative to those of the control group), as indexed by plasma corticosterone levels.

Sex differences in biobehavioral responses to conflict in a taste aversion paradigm.

A conditioned aversion to a novel milk solution was produced, and animals were then reexposed to milk while nondeprived (low conflict) or following a 72-hr food and water deprivation regimen (high conflict). No sex differences occurred if animals were nondeprived throughout testing. However, if deprived during the interval between conditioning and reexposure, sex differences in both behavior and adrenocortical responses occurred: (1) Presession corticoid levels of females were higher than those of males. (2) On the first reexposure day, females showed a suppression of plasma corticosterone below presession levels, while males maintained elevated or increased corticosterone levels. (3) On the second reexposure day, when no longer deprived, males showed a marked suppression of intake compared to females, and females subsequently recovered to pretoxicosis intake levels faster than males. (4) Gonadectomy eliminated these sex differences. While ovariectomized females continued to resemble intact females in both behavioral and hormonal responses, castrated males exhibited a corticoid suppression on the first reexposure day, and subsequently recovered to pretoxicosis intake levels at the same rate as females.

Long-term effects of early iron deficiency on consummatory behavior in the rat.

Two experiments were designed to investigate the effects of early iron deficiency on consummatory behavior in the adult rat. In experiment 1, animals were placed in a novel chamber, either with or without water available. Although there were no effects of iron deficiency per se, the data suggested that decreased caloric intake experienced early in life may have different long-term consequences for males and females. While ad lib control males, and females in all diet conditions, exhibited less elevation of plasma corticosterone when water was available in the novel chamber, calorically restricted males appeared unable to use the cues or reinforcement provided by consummatory behavior to reduce arousal. In Experiment 2, a conditioned taste aversion situation involving conflict, we were able to separate effects due to early iron deficiency from those due to early caloric restriction. When reexposed to milk, calorically restricted (weight control) males exhibited an attenuated plasma corticoid response, compared to that of ad lib control males, while weight control females resembled ad lib control females in their response. Thus, as in Experiment 1, early caloric restriction affected males more than females. Early iron deficiency, however, markedly altered pituitary-adrenal responsiveness in both males and females. Not only was the response to reexposure completely reversed in rehabilitated males and females, but also, the corticoid response to deprivation was increased in rehabilitated males and decreased in rehabilitated females. Taken together with previous data, these results suggest that early iron deficiency alters both behavioral and physiological arousal or responsiveness, and may do so differentially in males than females.

Schedule-induced polydipsia suppresses pituitary-adrenal activity in rats.

The effects of schedule-induced polydipsia (SIP) on pituitary-adrenal activity, as indicated by plasma levels of corticosterone, were examined in a series of experiments. Male (Experiments 1 and 3) and female (Experiment 2) rats were reduced to 80% of their free-feeding weight and given daily sessions on an intermittent-feeding schedule (fixed time of 60 sec). Half of the subjects in each experiment had water available during experimental sessions and the other half did not. Animals with water available in the experimental chamber exhibited SIP in all three experiments. In Experiment 1, blood samples were collected following (a) food consumption in the home cage, (b) a session on FT 60 sec, and (c) a session with pellets available in a cup in the experimental chamber. In Experiment 2, blood samples were taken prior to and following an FT 60-sec session, and following a session with pellets available in a cup in the chamber. In Experiment 3, pre- and postsession samples were obtained as in Experiment 2 (Part A). Subsequently, the opportunity to drink during sessions was removed, and the effect on corticoids was examined (Part B). The results indicate that (a) schedule-induced drinking suppresses pituitary-adrenal activity, (b) corticoid suppression may become a conditioned response to drinking in the chamber, and (c) corticoids return to presession levels following removal of water from the chamber. In view of these findings, it is hypothesized that SIP may serve an arousal-reducing role in intermittent-feeding situations.