H.D. Veldhuis

ACTH, cyclic nucleotides, and brain protein phosphorylation in vitro

Endogenous phosphorylation of proteins from rat brain synaptosomal plasma membranes was studied in vitro. Cyclic AMP (cAMP) markedly stimulated32P incorporation in three protein bands with molecular weights of 75,000, 57,000, and 54,000, respectively. The effect of the behaviorally active peptide ACTH1–24 on this endogenous phosphorylation in vitro was studied using peptide concentrations from 10−10 to 10−4 M. In a number of protein bands, a biphasic effect of ACTH1–24 was observed: in concentrations of 10−4–10−5 M, a reduced amount of32P was found; in concentrations of 10−6–10−7 M, hardly any effect could be detected, whereas consistently at concentrations around 10−8 M, a significant decrease was again observed. The phosphoprotein bands affected by in vitro addition of ACTH1–24 were of a smaller molecular weight than those affected by in vitro addition of cAMP.

Glucocorticoids facilitate the retention of acquired immobility during forced swimming

The adrenalectomy-induced decrease in the level of immobility during a 5 min retest period in the Porsolt swimming test could be reversed by glucocorticoids administered s.c. 15 min after the initial forced swimming exposure. The synthetic glucocorticoids dexamethasone and RU 28362 were active in the microgram dose range while corticosterone was only active at a 500 times higher dose. Aldosterone and progesterone were both ineffective. Treatment of adrenalectomized rats with the synthetic antiglucocorticoid RU 38486 prior to dexamethasone administration dose dependently blocked the effect of the glucocorticoid. Intact rats treated with the antiglucocorticoid RU 38486 prior to the initial forced swimming exposure behaved like adrenalectomized animals during the 5 min retest period. Removal of the adrenal medulla only temporarily impaired swimming behavior. It is concluded that intact adrenocortical secretion of glucocorticoids is sufficient for retention of acquired immobility during forced swimming.

Adrenal Steroids and Behavioral Adaptation: Relationship to Brain Corticoid Receptors

The words adrenocortical hormones and adaptation have been linked for almost 40 years. The concept as formulated by Selye (1950) related the adrenal Steroids to physiological, primarily peripheral adaptive processes. It has, however, become evident that the brain is an important target organ for corticosteroids. Psychological changes, such as mood alterations and psychotic reactions, have been frequently observed in patients receiving corticosteroid therapy (see von Zerssen 1976). Although the majority of reports hardly exceeded anecdotal level, therapeutic hopes were stirred for the mentally ill. The hopes and wishes of the early 1950s remained unfulfilled. Regular basic research concerning the pituitary-adrenal system and behavior was not started until the 1960s. Concepts which stemmed from this research led to the endocrine view of brain function and dysfunction held today. In this paper we want to review the past (not seeking completeness), to describe the present State of research, and to provide our view of the future of corticosteroid-brain interactions in understanding the endocrine modulation of behavioral adaptation.

Spatial learning and the hippocampal corticosterone receptor system of old rats: effect of the ACTH4-9 analogue ORG 2766.

Old (26 months) and young (6 months) male Wistar rats were treated chronically for 2 weeks with ORG 2766 or with vehicle, delivered via subcutaneously implanted minipumps (0.5 microgram peptide/0.5 microliter/h). Learning of a spatial task was not impaired in the old animals, except for one measure, i.e. the latency to find the goal box. In neither age group did ORG 2766 influence behavioral performance. The number of corticosterone receptor sites was decreased in the hippocampus of senescent rats, but restored to the level observed in young rats following ORG 2766 treatment. It is concluded that the number of hippocampal corticosterone receptor sites is a sensitive index of brain aging and effectiveness of ORG 2766.

Hippocampal corticosterone receptors and novelty-induced behavioral activity: Effect of kainic acid lesion in the hippocampus

Rats were injected bilaterally in the dorsal and ventral hippocampus with kainic acid (KA) or with artificial CSF and their behavior and brain corticosterone (B) receptor systems were studied. The hippocampal KA injection destroyed part of the pyramidal neurons and of the dentate gyrus neurons. These neurons contain a receptor system for B. At 2 weeks after the KA lesion this B receptor system displays an increase in apparent maximal binding capacity (Bmax) of approximately 25%. The compensatory increase in B receptor concentration is reflected in an increased uptake of [3H]B in cell nuclei of hippocampal slices incubated in vitro with saturating concentrations of the steroid. Administration of a tracer dose of [3H]B shows that labelled steroid can enter in vivo the cell nuclear compartment of the KA-lesioned lobe. The role of B was investigated on novelty-induced behavioral activities of KA-lesioned and sham-lesioned animals in a large open and a small closed field at 10 days after bilateral adrenalectomy (ADX) or sham-ADX which is 14 days after the (sham) lesion. B (300 micrograms/kg rat) was administered s.c. 1 h prior to the test. KA lesion resulted in an increase in exploratory activity and a reduction in grooming and immobility. After ADX the effect of KA on exploration was reduced in the 5 min open field and abolished in the 30 min closed field. ADX animals displayed more grooming behavior (closed-field). B replacement of ADX rats reinstated the exploratory hyperactivity of KA-lesioned rats. On some components of the behavior such as ambulation in open-field and locomotion in closed field, there was even a larger responsiveness to B in the KA-lesioned rats than in the control animals. It is concluded that (1) after KA lesion of receptor containing neurons, the remaining tissue displays a compensatory increase in number of B receptor sites; (2) B is required for full expression of exploratory activity of rats with or without KA lesions; (3) the KA-lesioned rats display a larger responsiveness to B; and (4) the increased number of B receptor sites may underlie the larger responsiveness to B.

Significance of ACTH4-10 in the control of hippocampal corticosterone receptor capacity of hypophysectomized rats

The effect of hypophysectomy on the number of corticosterone receptor sites was investigated in three rat brain regions and was compared with the effect of long-term adrenalectomy. Subsequently, the effect on receptor capacity was measured after the hypophysectomized rats had received as substitution therapy ACTH1-24 and smaller peptide fragments lacking corticotropic activity. All rats (sham and hypophysectomy) were adrenalectomized 24 h prior to sacrifice for depletion of endogenous adrenal hormones and replacement therapy was discontinued at that time. Extensive perfusion with saline was carried out at the time of sacrifice. 3H-corticosterone binding was measured in cytosol by means of an in vitro assay. 2 weeks after hypophysectomy, the apparent maximal binding capacity (Bmax) of the corticosterone receptor system was increased by 60, 36 and 72% in hippocampus, hypothalamus and septum, respectively. The increase in Bmax in the hippocampus of hypophysectomized rats was 30% higher than the increase in animals adrenalectomized 2 weeks previously. Replacement with ACTH1-24 markedly decreased the binding capacity in all brain regions investigated. Replacement with the behaviorally active ACTH4-10 sequence reduced the number of corticosterone receptor sites in the hippocampus by 21%, while the behaviorally inactive ACTH11-24 sequence was ineffective. Des-glycinamide-arginine-vasopressin was also ineffective. There were no alterations in binding affinity for corticosterone in hippocampal cytosol after the surgical procedures or after the different replacement therapies. It is concluded that the neurotropic ACTH4-10 sequence reduces the number of corticosterone receptor sites in the hippocampus of the hypophysectomized rat. The action of ACTH4-10 was specific for the hippocampus and was not observed in other brain regions or plasma transcortin.

Corticosteroid effects on morphine-induced antinociception as a function of two types of corticosteroid receptors in brain

The antinociceptive effect of parenterally and intracerebroventricularly injected morphine and beta-endorphin in adrenalectomized rats and in adrenalectomized rats treated with adrenal steroids was examined employing the hot-plate method. (1) Adrenalectomy sensitized the rats to an analgesic effect of morphine and beta-endorphin. (2) Replacement therapy (chronic and acute) with corticosterone, dexamethasone or RU 28362 (glucocorticoid receptor agonist) effectively reversed the increase in the sensitivity to the analgesic effect of peripherally injected morphine (5 mg/kg i.p.) induced by adrenalectomy to the level of sham-operated animals. Glucocorticosteroids administered to non-adrenalectomized rats did not change the sensitivity to morphine. (3) Corticosterone had a biphasic, dose-dependent effect; the most significant attenuation of the hypersensitivity to morphine-induced antinociception in adrenalectomized rats was achieved after 0.01 mg and after 10 mg (per kg b.w.). Doses of corticosterone of 0.005 mg/kg and in a range of 0.05-0.30 mg/kg were ineffective. (4) Corticosterone in a dose of 0.01 mg/kg (s.c.) had suppressant effects on the adrenalectomy-induced increase in the sensitivity to antinociception induced by morphine when given prior to morphine (60, 30 and 5 min) as well as after the injection of morphine (before the first and the second testing on the hot-plate, 15 and 5 min, respectively). (5) Intracerebroventricularly (i.c.v.) injected morphine and beta-endorphin also displayed the hypersensitivity to the analgesic effect in adrenalectomized rats which in both cases could be suppressed by 0.01 mg/kg of corticosterone given subcutaneously 5 min prior to administration of the opiate.(ABSTRACT TRUNCATED AT 250 WORDS)

Behavioural actions of vasoactive intestinal peptide (VIP)

The effect of vasoactive intestinal peptide (VIP) was studied on fear-motivated behaviours, exploration of a novel environment and on novelty and ACTH-induced grooming. VIP was administered via a plastic cannula into the lateral ventricle. Retention of a step-through passive avoidance task was inhibited by 10 and 30 ng VIP injected 1 hour before the retention test. Extinction of pole-jumping active avoidance behaviour was facilitated by 10 and 100 ng VIP. Mild effects were observed in an open field test on exploration and grooming activity. In conclusion, VIP produces inhibitory effects on fear-motivated behaviours.

Glucocorticoids modulate the response of ornithine decarboxylase to unilateral removal of the dorsal hippocampus

The effect of unilateral removal of the dorsal hippocampus and of glucocorticoid administration was measured on the activity of ornithine decarboxylase (ODC) in the remaining contralateral hippocampus lobe. Unilateral hippocampectomy (Hx) resulted in a rapid rise of ODC activity in the contralateral lobe. The effect on ODC was maximal at 6 h after surgery and lasted two days. In the absence of the adrenals the effect of Hx on the enzyme was more potent and more prolonged. Elevated ODC activity was still detectable at 5 days after surgery, but not at 10 days. Chronic replacement with dexamethasone (DEX) offered in drinking water decreased the Hx-induced ODC response of ADX rats at 3 days after surgery to the level of enzyme activity observed in the S-ADX Hx subject. The effect of the steroid seemed related to the extent of occupation of the pool of glucocorticoid receptor sites in cytosol of rat hippocampus. In contrast, a single injection of a high dose of DEX to Hx-ADX animals at 3 days after surgery increased ODC activity in addition to the lesion-induced ODC in the contralateral lobe. It is concluded that after unilateral removal of the dorsal hippocampus ODC is a biochemical marker for cellular responses taking place in the contralateral lobe. Glucocorticoids modulate the lesion-induced ODC response.

The hippocampal corticosterone receptor system of the homozygous diabetes insipidus (Brattleboro) rat

The binding of [3H] corticosterone to hippocampal cytosol receptors of Brattleboro rats homozygous for diabetes insipidus (Ho-Di) and of normal Brattleboro rats (Ho-No) was investigated at 24 h after removal of the adrenals. The apparent maximal binding capacity of the Ho-Di hippocampal corticosterone receptor system was about 30% less than that of the Ho-No rats. Substitution of the vasopressin deficient rats with 1E pitressin tannate in oil partially restores the hippocampal corticosterone receptor level towards that of the control animals.