Hypothalamic-pituitary-adrenal-cortical activity in anorexia nervosa and bulimia

We studied hypothalamic-pituitary-adrenal-cortical (HPA) activity in nine underweight women with anorexia nervosa, 12 women of normal body weight with bulimia, and nine control subjects. The measures of HPA activity were the pattern of plasma cortisol secretion over 24 hr and the responses of plasma cortisol to dexamethasone suppression and to low dose ACTH stimulation. The patients with anorexia nervosa had significantly elevated 24 hr concentrations of plasma cortisol compared to the controls and showed significantly less cortisol suppression following dexamethasone. There was no difference between patients with anorexia nervosa and controls in the rise in plasma cortisol following ACTH. On most measures of HPA activity, the normal weight patients with bulimia were indistinguishable from the controls. These results suggest that HPA activity is normal in most patients of normal body weight with bulimia and that the psychological and behavioral disturbances common to both anorexia nervosa and bulimia are, in the absence of significant weight loss, insufficient to produce major alterations in HPA activity.     

Lymphocyte glucocorticoid receptor binding in depressed patients with hypercortisolemia

Despite elevated levels of serum and urinary cortisol, patients with depressive illness manifest none of the clinical stigmata of glucocorticoid excess. This hypercortisolemia in the absence of clinical effects suggests a state of hormone resistance and could be mediated by alterations in the glucocorticoid receptor. Earlier studies have shown that small doses of glucocorticoids cause a decrease in glucocorticoid receptor binding in normal human lymphocytes. White cells from depressed patients with significant hypercortisolemia would be expected to show a similar change in receptor concentration if peripheral tissues are adequately exposed to and sensitive to the hormone. In this study we compared the binding of [³H]dexamethasone to lymphocytes from normal subjects and depressed patients with hypercortisolemia. Lymphocytes from normal subjects had a mean receptor concentration of 10.2 ± 0.66 fm/10⁶ cells (S.E.M.) and a dissociation constant of 4.8 ± 0.47 nM. Lymphocytes from depressed patients with abnormal 0800 h serum cortisol after dexamethasone had a mean receptor concentration of 8.8 ± 0.75 fm/10⁶ cells, which was not significantly different from that in lymphocytes from normal subjects or from depressed subjects with normal post-dexamethasone cortisol levels (9.4 ± 0.95 fm/10⁶ cells). Lymphocytes from depressed patients with elevated urinary free cortisol excretion (UFC) also had normal receptor concentration and binding affinity for dexamethasone. The lack of a change in lymphocyte glucocorticoid receptor concentration in the presence of cortisol excess suggests the possibility that hypercortisolemia in depressive illness represents a state of peripheral glucocorticoid resistance.     

Glucocorticoid binding sites in human temporal cortex

This work describes the presence of glucocorticoid binding sites in human temporal cortex obtained following partial lobectomy in two epileptic patients. Using [³H]dexamethasone as radioligand and cold cortisol or RU 28362 as competitor we found an apparent K_d of 2.8 nM with a B_max of approximately 34 fmol/mg protein. The order of potency of various unlabeled steroids to compete for [³H]dexamethasone binding was as follows: RU 28362 = RU 38486 = cortisol = dexamethasone > progesterone > spironolactone > estradiol. These data provide evidence for an intracellular mechanism by which circulating glucocorticoids might regulate neuronal function in the human cortex.     

Desipramine treatment has minimal effects on the brain accumulation of glucocorticoids in P-gp-deficient and wild-type mice

Hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis in patients with depression can be reduced by antidepressants, which are thought to improve endogenous glucocorticoid-mediated negative feedback. A proportion of peripherally released glucocorticoids need to enter brain tissue, protected by the blood-brain barrier (BBB), in order to achieve this negative feedback effect at the level of the central nervous systems (CNS). The multidrug resistance transporter P-glycoprotein (P-gp) has been shown to actively transport glucocorticoid hormones and has been implicated in the regulation of glucocorticoid access to the CNS. Using an in situ brain/choroid plexus perfusion method, we tested the hypothesis that the antidepressant desipramine increases glucocorticoid accumulation in the mouse brain by inhibiting P-gp, following either chronic treatment (8 days, 20 mg/kg/day, IP) or acute administration (20 min brain perfusion in the presence of either 0.9 μM or 10 μM desipramine). Contrary to our hypothesis, chronic treatment with desipramine did not affect the accumulation of [3H]dexamethasone in any sample compared to saline-treated mice. Acute desipramine had limited and variable effects on glucocorticoid accumulation in the CNS, with accumulation of [3H]dexamethasone increased in the cerebellum, accumulation of [3H]cortisol reduced in the frontal cortex, hypothalamus, and cerebellum, and accumulation of [3H]corticosterone (the endogenous glucocorticoid in rodents) not affected. Overall, under the conditions tested, these results do not support the hypothesis that treatment with desipramine can inhibit P-gp at the BBB and subsequently increase the accumulation of glucocorticoids in the brain.     

Glucocorticoid receptors of mononuclear leukocytes from myasthenia gravis patients

The present study was performed to analyse glucocorticoid receptor (GR) binding in peripheral blood mononuclear leukocytes (MNL) from 39 myasthenia gravis (MG) patients (unoperated patients (n = 13), thymectomized patients (n = 14) and patients receiving glucocorticoids: thymectomized (n = 11) and unoperated (n = 6]. A whole cell binding assay with 3(H) dexamethasone was used. GR mean values were significantly higher in the MNL of MG patients (thymectomized or not) not receiving glucocorticoid than in the MNL of healthy donors. Affinity was within the normal range. Sex, age or clinical forms of illness did not influence the results. In patients receiving prednisone (Pd) the GR values were significantly lower than in MG patients without Pd therapy, independent of Pd dose or time of administration. No differences in receptor binding between normal subjects and MG patients receiving Pd have been found.     

Prolactin responses in dexamethasone suppression test in patients with anorexia nervosa

In anorexia nervosa (AN), abnormalities are present in the hypothalamic-pituitary-adrenal axis, but the prolactin (PRL) response to dexamethasone suppression test (DST) has not yet been studied. In order to study the interrelationships between the various endocrine abnormalities, we investigated the responses of PRL and cortisol to DST (1 mg of dexamethasone at 2300) in AN patients. The subjects were 12 female inpatients with AN and 8 age- and sex-matched healthy controls. The percentage suppression and absolute change in PRL levels before and after dexamethasone administration were significantly different in the 2 groups. In the control group PRL levels suppressed to 36.5 +/- 3.7% of basal, while AN levels declined to 79.4 +/- 8.9% of basal. When the percentage suppression of PRL was compared between patients with and without cortisol suppression, the mean PRL level was 68.9 +/- 7.8% of the basal level for the cortisol-suppressed patients and 100.4 +/- 19.1% for the nonsuppressed patients. Hence in both groups, the percentage PRL suppression was significantly reduced compared with the control group, and indeed nonexistent in cortisol-nonsuppressed patients. The finding that there was less PRL suppression in the cortisol-suppressed patients than in the controls suggests that, in AN, there may be an abnormality in PRL secretion not related to the hypothalamic-pituitary-adrenal axis. Further work is needed to distinguish between the PRL response to stress and potential hypothalamic abnormality.     

The Dexamethasone Suppression Test for Japanese with Eating Disorders

Abstract: A one-mg oral overnight dexamethasone suppression test (DST) was conducted on 22 inpatients with eating disorders. To confirm that the dexamethasone tablets had been ingested, we measumd the plasma concentrations of dexamethasone the next morning (at 0900 hr after DST). The diagnosis of anorexia nervosa and bulimia was made according to the criteria for DSM-III, respectively. Of the 22 patients with eating disorders, 16 satisfied the criteria for anorexia nervosa and 6 for bulimia. The DST was carried out within 2 weeks of hospitalization on each patient. The subjects were given 1 mg of dexamethasone in the evening (at 2300 hr) and blood samples were collected the following day (at 0900, 1600 and 2100 hr, respectively). The plasma cortisol and dexamethasone levels were concurrently determined by RIA. The criterion for non-suppression was a failure to suppress the plasma cortisol levels below 5.0 μg/dl in any one of the three samples. All but one patient with bulimia had ingested the dexamethasone. Thirteen (62%) of 21 patients with eating disorders were nonsuppressors. We found a significant positive correlation between the plasma cortisol levels at 1600 hr or 2100 hr and a decrease in ideal body weight (n = 16, r = 0.613, p < 0.05; r = 0.75, p < 0.01, respectively) and a significant inverse relationship between the plasma dexamethasone levels at 0900 hr and the plasma cortisol levels at 1600 hr was recognized (n = 21, r - 0.631, p < 0.01). These results suggest that the blood dexamethasone levels as well as body weight might contribute to the abnormalities of DST seen in patients with eating disorders.     

The dexamethasone suppression test for Japanese with eating disorders

A one-mg oral overnight dexamethasone suppression test (DST) was conducted on 22 inpatients with eating disorders. To confirm that the dexamethasone tablets had been ingested, we measured the plasma concentrations of dexamethasone the next morning (at 0900 hr after DST). The diagnosis of anorexia nervosa and bulimia was made according to the criteria for DSM-III, respectively. Of the 22 patients with eating disorders, 16 satisfied the criteria for anorexia nervosa and 6 for bulimia. The DST was carried out within 2 weeks of hospitalization on each patient. The subjects were given 1 mg of dexamethasone in the evening (at 2300 hr) and blood samples were collected the following day (at 0900, 1600 and 2100 hr, respectively). The plasma cortisol and dexamethasone levels were concurrently determined by RIA. The criterion for non-suppression was a failure to suppress the plasma cortisol levels below 5.0 micrograms/dl in any one of the three samples. All but one patient with bulimia had ingested the dexamethasone. Thirteen (62%) of 21 patients with eating disorders were nonsuppressors. We found a significant positive correlation between the plasma cortisol levels at 1600 hr or 2100 hr and a decrease in ideal body weight (n = 16, r = 0.613, p less than 0.05; r = 0.75, p less than 0.01, respectively) and a significant inverse relationship between the plasma dexamethasone levels at 0900 hr and the plasma cortisol levels at 1600 hr was recognized (n = 21, r = 0.631, p less than 0.01). These results suggest that the blood dexamethasone levels as well as body weight might contribute to the abnormalities of DST seen in patients with eating disorders.     

Development of neuronal responses in cat posteromedial lateral suprasylvian visual cortex

Exchange assays have often been used to quantitate steroid receptors when endogenous ligands are present; however, there are no reports of their successful application to activated glucocorticoid-Type II receptor complexes. In addition to investigating the reasons for this failure, the present study also examined the effects of progesterone on glucocorticoid dissociation from, and reassociation with unactivated and activated Type II receptors. Molybdate-stabilized brain cytosol from adrenal-ovariectomized mice was incubated with [3H]dexamethasone ( +/- [1H]DEX) for 40 h at 0 degree C. Afterwards free steroid was removed on Sephadex G-25 columns in the presence (unactivated receptors) or absence (activated receptors) of molybdate. Activation, as measured by DNA-cellulose binding, was achieved by incubating molybdate-free cytosol at 22 degrees C for 20 min followed by G-25 filtration in the presence of molybdate. The rates of dissociation and reassociation were then measured by incubating cytosol with [1H]triamcinolone acetonide (TA) or [3H]TA ( +/- [1H]TA) at 12 degrees C. An exchange assay was also employed in which cytosol was incubated first with [1H]DEX for 40 h at 0 degree C followed by bound-free steroid separations and 12 degrees C incubations with [3H]TA ( +/- [1H]TA). Both approaches revealed that even though activation reduced the rate of DEX dissociation from Type II receptors by 40%, it eliminated the ability of the newly unoccupied receptors to rebind glucocorticoid. Adding [1H]progesterone to occupied receptor preparations increased dissociation rate constants by nearly 3-fold, for both unactivated and activated Type II receptors. Since [1H]TA failed to prevent this effect, progesterone appears to act at an allosteric site(s) which cannot be occupied by glucocorticoids. Exchange assays revealed that progesterone-facilitated dissociation increased the rate of glucocorticoid rebinding to unactivated, but not activated Type II receptors. These results suggest that spontaneous and progesterone-facilitated termination of glucocorticoid genomic actions could be mediated by steroid dissociation since unoccupied activated Type II receptors do not rebind agonist steroid.     

Basal activity of the hypothalamic-pituitary-adrenal axis and cognitive function in anorexia nervosa

Anorexia nervosa is associated with abnormalities in neuroendocrine function including sustained hypercortisolism, which has been shown elsewhere to be associated with impairment of function in learning, memory and attention. Cognitive impairment has also been observed in anorexia nervosa. These effects may be mediated in part through cortisol effects on the hippocampus, which is dense with glucocorticoid receptors. We investigated the association between cortisol levels and cognitive function in anorexia nervosa by measuring both 24-hour urinary cortisol counts and performance on tasks of learning, memory and attention in patients suffering from the disorder. Cortisol secretion was shown to be significantly higher in the patient group than in a matched control group and patients were also shown to be impaired in memory and attention. However, no correlations were found between the cognitive deficits and cortisol measures. It is suggested that more sensitive profiling of cortisol levels throughout the circadian cycle may be useful in future studies of cognitive function in anorexia nervosa. Received: 14 April 1999 / Accepted: 29 October 1999     

Fetal dexamethasone exposure sensitizes neonatal rat brain to hypoxia: effects on protein and DNA synthesis.

Fetal exposure to glucocorticoids is known to produce long-term alterations in cell development within the central nervous system. The current study examines whether some of the adverse effects of prenatal dexamethasone treatment on brain development represent sensitization to hypoxia-induced damage. Pregnant rats were given 0.2 or 0.8 mg/kg of dexamethasone on gestational days 17, 18 and 19 and their offspring were challenged by exposure to 7% O2 on postnatal days 1 and 8. In control rats at 1 day of age, hypoxia evoked an acute decrease in protein synthesis, assessed by [3H]leucine incorporation, in both the midbrain + brainstem and forebrain. The decrease was also seen in animals receiving the low dose of dexamethasone, but was of smaller magnitude in the midbrain + brainstem than in the control cohort. At the higher dose of dexamethasone, hypoxia failed to evoke a decrease in protein synthesis; instead, protein synthesis was significantly increased. By 8 days of age, the animals receiving the lower dose of dexamethasone also displayed the anomalous increment in [3H]leucine incorporation during hypoxic challenge, whereas the effect in the high dose group was less notable. Similarly, parallel examination of incorporation of [3H]thymidine into DNA on postnatal day 1 indicated that control animals would reduce their macromolecule synthetic rate in a hypoxic environment, but that animals exposed to the high dose of dexamethasone would not; unlike the case with protein synthesis, however, the dexamethasone group never showed an increase in DNA synthesis during hypoxia. By 8 days of age, the interaction between the high dose of dexamethasone and hypoxia was no longer apparent for DNA synthesis.2     

Partial 11 beta-hydroxylase activity suppression after dexamethasone in patients with major depression

Eleven beta-hydroxylase activity was assessed by measuring the cortisol to 11-deoxycortisol ratio in 20 control subjects, 38 patients with major depression, and five patients with Cushing's disease before and after 1 mg of dexamethasone. The mean levels of 11 beta-hydroxylase activity did not differ among groups before dexamethasone. After dexamethasone patients with Cushing's disease showed a nonsignificant increase in 11 beta-hydroxylase activity while patients with major depression and controls subjects both showed a decrease. Endogenous depressive patients were no more likely to show high 11 beta-hydroxylase activity than neurotic depressive patients; however, depressed patients with cortisol nonsuppression after dexamethasone were. Post-dexamethasone 11 beta-hydroxylase activity is positively correlated with age in both control subjects and patients with depression.     

Effects of glucocorticoids on declarative memory function in major depression.

BACKGROUND: Major depression has been associated with hypercortisolemia in a subset of patients with depression. Administration of exogenous cortisol and other glucocorticoids to healthy human subjects has been observed to result in a transient impairment in verbal declarative memory function. The purpose of this study was to assess the effects of the glucocorticoid, dexamethasone, on verbal declarative memory function in patients with untreated unipolar major depressive disorder (MDD). METHODS: Fifty two men and women with (n = 28) and without (n = 24) MDD received placebo or dexamethasone (1 mg and 2 mg on 2 successive days) in a double-blind, randomized fashion. Declarative memory was assessed with paragraph recall at baseline (day 1) and day 3. RESULTS: There was a significant interaction between diagnosis and drug (dexamethasone vs. placebo) on paragraph recall. In the healthy subjects, memory improved from baseline to day 3 with placebo and was unchanged with dexamethasone, whereas in MDD patients memory function showed a pattern of decreasing with placebo and improving with dexamethasone from baseline to day 3. CONCLUSIONS: These findings are consistent with an altered sensitivity of declarative memory function in MDD to regulation by glucocorticoids. Possible explanations of the findings include alterations in glucocorticoid receptors in the hippocampus or other brain regions mediating declarative memory, or differential sensitivity to dexamethasone-induced reductions in cortisol, in patients with MDD.     

Enhanced cortisol suppression in eating disorders with impulsive personality features

Evidence of both blunted and enhanced cortisol suppression with the dexamethasone test (DST) is available in eating disorders (ED), suggesting that different subtypes of ED might be characterized by distinct neurobiological stress response dysfunctions. Other evidence indicates that ED patients with impulsive clinical features might have enhanced cortisol suppression similar to patients with impulsive personality disorders. A group of 52 patients with restrictive anorexia, binge eating-purging anorexia and bulimia nervosa were studied with a very low dose (0.25 mg) dexamethasone test and measures of phenomenology, personality and impulsivity. Patients with bulimic symptoms had significantly higher rates of cortisol suppression than controls and than restrictive anorectic patients. Percent cortisol suppression showed a strong and significant correlation with the patient's score on the Barratt Impulsiveness Scale. A hypersensitive cortisol response to dexamethasone, which might reflect hypothalamic-pituitary-adrenal axis dysfunctions might be specifically associated with impulsive subtypes of eating disorders.     

Daily profiles of melatonin, cortisol and gonadotropins in 8 adolescents with anorexia nervosa

From a biological study of 8 patients with anorexia nervosa and an analysis of the daily profiles of gonadotropins, cortisol and melatonin, the authors discuss the possible relationship between anorexia nervosa and affective disorders. At the initial phase of the illness, plasmatic levels of FSH, LH and oestradiol were very low. 24-hour plasma cortisol values were comparable with those of a depressive population; when weight loss was equal to or higher than 25% of the initial weight, there was no suppression by dexamethasone. The daily profile of melatonin was maintained, with melatonin plasma levels significatively higher than in a control group of depressed patients.     

Disturbances in the hypothalamo-pituitary-adrenal and other neuroendocrine axes in bulimia

Disturbances in the hypothalamo-pituitary-adrenal (HPA) and other endocrine axes were assessed in 24 women with bulimia and healthy controls. Overnight blood samples for measuring nocturnal plasma cortisol, prolactin (PRL), growth hormone (GH), luteinizing hormone (LH), and follicle stimulating hormone (FSH) were obtained at 30-min intervals. A 1.5 mg dexamethasone suppression test (DST) and a TRH-test were performed. Patients were monitored closely while their nutritional intake was recorded over 21 days. Compared with healthy controls, nocturnal cortisol plasma levels were not elevated in the bulimics. There was a trend toward insufficient cortisol suppression in the DST in patients with bulimia, which was most pronounced in patients with signs of restricted caloric intake. Plasma dexamethasone levels were significantly reduced in bulimics compared with healthy controls. There was a trend for blunted thyrotropin stimulating hormone (TSH) responses to thyrotropin releasing hormone (TRH) in bulimia. The prolactin response to TRH was significantly reduced in bulimics with a history of anorexia nervosa. Plasma LH and plasma FSH were significantly reduced in bulimics with signs of reduced caloric intake [low T3, high levels of beta-hydroxy-butyric acid (BHBA), reduced daily caloric intake, high number of fasting days] as compared with healthy controls. Bulimics with high BHBA levels had significantly reduced nocturnal prolactin plasma levels. Results show that multiple neuroendocrine disturbances exist in bulimia in a milder form than in anorexia nervosa. Evidence for the impact of caloric intake on endocrine functions is presented. Endocrine dysfunctions in our bulimic sample did not show a positive association with the presence of depressive symptoms.     

The dexamethasone suppression test: importance of dexamethasone concentrations

Plasma dexamethasone concentrations and cortisol response to dexamethasone were measured in 29 normal healthy volunteers, 23 depressed patients, and 10 patients with anorexia nervosa at 4:00 PM postdexamethasone. In each of the 3 groups, nonsuppressors had lower dexamethasone concentrations than suppressors. Of the subjects with plasma dexamethasone at or below 0.7 ng/ml, a significantly higher proportion (48%) were nonsuppressors compared to the proportion above 0.7 ng/ml (14%), all of whom were patients. Plasma dexamethasone concentrations in a subgroup of depressed nonsuppressors were high (mean 1.35 ng/ml), whereas the remainder were low (0.42 ng/ml) and were similar to the normal nonsuppressors (0.35 ng/ml), suggesting different mechanisms for nonsuppression in the subgroups. Plasma dexamethasone concentrations were similar in nonendogenous and endogenous depressives, in men and women, and in medicated and drug-free patients. None of the variables of age, weight, history of weight loss, Hamilton depression rating score, predexamethasone cortisol, or postdexamethasone cortisol were significantly correlated with plasma dexamethasone, except for body weight and a history of weight loss in the depressed group only. Mean plasma dexamethasone concentrations increased significantly from week 1 to week 2 in 7 depressed patients, whereas plasma cortisol decreased; however, the relationship between dexamethasone and cortisol varied considerably for individual patients.     

Rat C6 glioma cells contain type I as well as type II corticosteroid receptors

Rat brain cytosol contains Type I corticosteroid receptors. Unlike Type II (glucocorticoid) receptors, Type I receptors have high affinity for the endogenous corticosteroids - aldosterone, deoxycorticosterone, and corticosterone - and much lower affinities for synthetic glucocorticoids. In the present study, we report that Type I corticosteroid receptors are present in C6 glioma cells. Type I receptors were identified in C6 cell cytosol and whole cells by the binding of [3H]aldosterone. The specific glucocorticoid RU 26988 was used to block Type II receptors. Measured in whole C6 cells, Type I receptors had a density of 2.1 +/- 1.1 fmol/10(6) cells and a dissociation constant (Kd) for [3H]aldosterone of 0.41 +/- 0.06 nM. The density of Type I receptors was only 2% of the density of Type II corticosteroid receptors (96 +/- 7 fmol/10(6) cells), measured in whole C6 cells by [3H]triamcinolone binding. The steroid specificity of glial cytosolic Type I receptors (deoxycorticosterone greater than corticosterone greater than aldosterone greater than dexamethasone greater than triamcinolone much greater than RU 26988) was identical to the steroid specificity of Type I receptors in rat brain cytosol. The potency of deoxycorticosterone was somewhat reduced when measured in whole cells. The steroid specificity of the Type I receptor differed markedly from that of the Type II (glucocorticoid) receptor (triamcinolone greater than dexamethasone greater than RU 26988 corticosterone greater than deoxycorticosterone greater than aldosterone). Since Type I receptors in the kidney mediate effects of aldosterone upon renal transport of sodium and potassium, it is proposed that glial Type I corticosteroid receptors may be involved in the regulation of glial ion transport.(ABSTRACT TRUNCATED AT 250 WORDS)     

Increased glucocorticoid production and altered cortisol metabolism in women with mild to moderate Alzheimer's disease.

BACKGROUND: Abnormalities at several levels of the hypothalamic-pituitary-adrenal axis, which may promote glucocorticoid dependent neurodegeneration, have been reported in patients with Alzheimer's disease. In this study we have explored the possibility that peripheral cortisol metabolism is enhanced and glucocorticoid production is increased compensatory in patients with mild to moderate Alzheimer's disease. METHODS: Urinary excretion of cortisol and its principal conjugated metabolites was studied in ten women with mild to moderate Alzheimer's disease, seven healthy elderly women and seven young women. RESULTS: There was an age-related fall in glucocorticoid production (median, 2009 [range 1828--4201] vs. 9315 [range 3613--16,244] microg/24 hours in elderly vs. young control subjects). A-ring metabolism (i.e., the irreversible inactivation of cortisol by 5 alpha- and 5 beta-reductases) was reduced in old age. However, patients with Alzheimer's disease showed persistence of cortisol metabolite excretion. Glucocorticoid production was significantly increased in patients with Alzheimer's disease versus healthy elderly control subjects (median, 7269 [range 6005-15,335] vs. 2009 [range 1828--4201] microg/24 hours), and patients with Alzheimer's disease had increased A-ring reduction of cortisol. CONCLUSIONS: An increased glucocorticoid production is an early feature of Alzheimer's disease and may be secondary to enhanced metabolic clearance of cortisol by A-ring reduction.     

Characterization of glucocorticoid receptors in whole and cellular subfractions of embryonic chick spinal cord

Steroid-specific binding sites for tritiated corticosterone have been localized, via autoradiography, to motoneurons in the lateral motor columns of the adult rat spinal cord. Binding sites in adult rat spinal cord have been characterized biochemically and shown to possess the properties of a putative glucocorticoid receptor. The presence of receptors for glucocorticoids in embryonic chick spinal cord was determined and their characterization undertaken as a prelude to the study of the functions under regulation by glucocorticoids during development. Assay conditions were defined and binding of [3H]dexamethasone [( 3H]Dex) to cytosols of 6- and 10-day embryonic spinal cord and cellular subfractions of 6-day spinal cord determined. Saturable, high-affinity binding of [3H]Dex to cytosols prepared from both whole 6- and 10-day spinal cords and cells of all 3 cellular subfractions of 6-day spinal cords was observed. The binding component in 10-day cytosols was (1) proteinaceous, as binding was eliminated by heating cytosols, and (2) a macromolecular species, as it displayed a sedimentation coefficient of 8S upon centrifugation in sucrose gradients. The putative receptor displayed binding specific for glucocorticoids in a competition assay, with the exception that some inhibition of binding by the androgen ligand, methyltrienolone (R1881) was observed. The binding affinity decreased as the values for KD increased from 3.4 +/- 0.9 nM on day 6 to 15.7 +/- 1.8 nM on day 10, while the values for Bmax increased from 270 to 855 fmol/mg protein over the same period.(ABSTRACT TRUNCATED AT 250 WORDS)