Melly S. Oitzl

Learning under stress: how does it work?

The effects of stress on learning and memory are not always clear: both facilitating and impairing influences are described in the literature. Here we propose a unifying theory, which states that stress will only facilitate learning and memory processes: (i) when stress is experienced in the context and around the time of the event that needs to be remembered, and (ii) when the hormones and transmitters released in response to stress exert their actions on the same circuits as those activated by the situation, that is, when convergence in time and space takes place. The mechanism of action of stress hormones, particularly corticosteroids, can explain how stress within the context of a learning experience induces focused attention and improves memory of relevant information.

Selective corticosteroid antagonists modulate specific aspects of spatial orientation learning

Receptors for mineralocorticoids (MRs) and glucocorticoids (GRs) display a high concentration and distinct distribution in the hippocampus. The effects of corticosteroids on behavior mediated by central MRs and GRs were assessed in rats. Spatial navigation is considered to be a sensitive measure for hippocampal functioning. Removal of circulating corticosteroids (via adrenalectomy) impaired spatial learning. In intact rats, blockade of central MRs and GRs by intracerebroventricular injection of selective MR and GR antagonists influenced different aspects of spatial learning. The analysis of the behavioral pattern revealed that treatment with the MR antagonist altered search-escape strategies in the water maze. The injection of the GR antagonist after training resulted in increased latencies to find the platform, which reflects the disturbed consolidation of spatial information. Corticosteroids affect in a differential and coordinated manner behavioral strategies and storage of spatial information.

Functional implications of brain corticosteroid receptor diversity

Summary1.Corticosteroids readily enter the brain and control gene expression in nerve cells via binding to intracellular receptors, which act as gene transcription factors. In the rat brain corticosterone binds to mineralocorticoid receptors (MRs) with a 10-fold higher affinity than to glucocorticoid receptors (GRs). As a consequence, these MRs are extensively occupied under basal resting conditions, while substantial GR occupation occurs at the circadian peak and following stress. Both receptors are colocalized in most, but not all, hippocampal neurons. In addition, some neurons contain aldosterone-selective MRs, if corticosterone is enzymatically inactivated. These aldosterone target neurons are presumably localized in the anterior hypothalamus, where they underlie central control of salt appetite and cardiovascular regulation.2.The data show that MR- and GR-mediated effects proceed in a coordinate and often antagonistic mode of action: (i) in hippocampus MR activation maintains excitability, while GR occupancy suppresses excitability, which is transiently raised by excitatory stimuli; (ii) central MRs participate in control of the sensitivity of the neuroendocrine stress response system, while GRs are involved in termination of the stress response: (iii) MRs in the hippocampus have a role in regulation of behavioral reactivity and response selection. GR-mediated effects facilitate storage of information.3.On the basis of these data, we propose that a relative deficiency or excess of MR- over GR-mediated neuronal effects may lead to a condition of enhanced or reduced responsiveness to environmental influences, alter behavioral adaptation, and promote susceptibility to stress. The findings may serve development of novel therapeutic strategies for treatment of stress-related brain diseases.

Interleukin-1β, but not interleukin-6, impairs spatial navigation learning

Abstract The effects of the cytokines interleukin-1 and -6 (IL1β, IL6; 100 ng) on spatial learning were examined in the Morris water maze. Intracerebroventricular injection of IL1 or IL6 before the training on day 1 did not influence the acquisition of spatial navigation. However, IL1 administered at 60 min, but not immediately before the training, resulted in impaired performance of spatial navigation the following day. In contrasts, IL6 administered at both times had no effect. In a second experiment the same doses of IL1 and IL6 increased the body temperature of rats in a time-related fashion. The temperature effect of IL1 developed after a delay of 120 min, while the IL6-effect was immediate. Comparable behavioral changes might accompany infections or inflammatory diseases and therapeutic cytokine administration.

The postnatal development of the hypothalamic-pituitary-adrenal axis in the mouse.

The main characteristic of the postnatal development of the stress system in the rat is the so‐called stress hypo‐responsive period (SHRP). Lasting from postnatal day (pnd) 4 to pnd 14, this period is characterized by very low basal corticosterone levels and an inability of mild stressors to induce an enhanced ACTH and corticosterone release. During the last years, the mouse has become a generally used animal in stress research, also due to the wide availability of genetically modified mouse strains. However, very few data are available on the ontogeny of the stress system in the mouse. This study therefore describes the postnatal ontogeny of peripheral and central aspects of the hypothalamic–pituitary–adrenal (HPA) axis in the mouse. We measured ACTH and corticosterone in blood and CRH, urocortin 3 (UCN3), mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) transcripts in the brain at postnatal days 1, 2, 4, 6, 9, 12, 14 and 16. Our results show that we can subdivide the postnatal development of the HPA axis in the mouse in two phases. The first phase corresponds to the SHRP in the rat and lasts from right after birth (pnd 1) until pnd 12. Basal corticosterone levels were low and novelty exposure did not enhance corticosterone or ACTH levels. This period is further characterized by a high expression of CRH in the paraventricular nucleus (PVN) of the hypothalamus. Expression levels of GR in the hippocampus and UCN3 in the perifornical area are low at birth but increase significantly during the SHRP, both reaching the highest expression level at pnd 12. In the second phase, the mice have developed past the SHRP and were now exhibiting enhanced corticosterone basal levels and a response of ACTH and corticosterone to mild novelty stress. CRH expression was decreased significantly, while expression of UCN3 and GR remained high, with a small decrease at pnd 16. The expression of MR in the hippocampus was very dynamic throughout the postnatal development of the HPA axis and changed in a time and subregion specific manner. These results demonstrate for the first time the correlation between the postnatal endocrine development of the mouse and gene expression changes of central regulators of HPA axis function.

The Effect of Corticosterone on Reactivity to Spatial Novelty is Mediated by Central Mineralocorticosteroid Receptors

Corticosterone, secreted by the adrenal glands, binds to central mineralocorticoid receptors with high affinity and to glucocorticoid receptors with a tenfold lower affinity. In previous studies we have shown that the selective activation of either mineralocorticoid receptors or glucocorticoid receptors exerts distinctly different behavioural effects. In this study we examined in particular the mineralocorticoid receptor‐mediated effect of corticosterone on the control of the behavioural response of male Wistar rats to spatial novelty. This analysis was based on our observation that in adrenal‐intact rats the presence of an object in the centre of an open field alters the time spent and distance walked in the centre compared to the peripheral area, i.e. the pattern of reactive locomotor activity is changed. Using this paradigm we found that 1 day after removal of the adrenals the rats increased their behavioural reactivity towards the object. Treatment of adrenalectomized rats with a low dose of corticosterone (50 μg/kg s.c.) 1 h prior to testing restored the behavioural reactivity to the level of sham‐operated, intact rats. Surprisingly, a high dose of corticosterone (1000 μg/kg s.c.) also increased the rats reactivity towards the object. The same high dose of corticosterone given to adrenal‐intact rats also increased behavioural reactivity. Pretreatment of these rats with an intracerebroventricular injection of the selective mineralocorticoid receptor antagonist RU28318 (100 ng/μl) prevented the corticosterone‐induced increase in behavioural reactivity, while the blockade of glucocorticoid receptors with the antagonist RU38486 (100 ng/μl) was not effective. Administration of the mineralocorticoid receptor antagonist without corticosterone to adrenal‐intact rats also increased behavioural reactivity, but this increase did not reach statistical significance. General locomotor activity was not affected by either treatment. In conclusion, we found a U‐shaped relationship between the pattern of behavioural reactivity in a novel environment and the circulating plasma corticosterone level. The response to spatial novelty appeared to be sensitive with respect to the activation and blockade of central, presumably hippocampal mineralocorticoid receptors.

Corticosteroids operate as a switch between memory systems

Stress and corticosteroid hormones are known to affect learning and memory processes. In this study, we examined whether stress and corticosteroids are capable of facilitating the switch between multiple memory systems in mice. For this purpose, we designed a task that allowed measurement of nucleus caudate-based stimulus–response and hippocampus-based spatial learning strategies. Naive mice used spatial strategies to locate an exit hole on a circular hole board at a fixed location flagged by a proximal stimulus. When the mice were either stressed or administered corticosterone before the task, 30–50% of the mice switched to a stimulus–response strategy. This switch between learning strategies was accompanied by a rescue of performance, whereas performance declined in the stressed mice that kept using the spatial strategy. Pretreatment with a mineralocorticoid receptor antagonist prevented the switch toward the stimulus–response strategy but led to deterioration of hippocampus-dependent performance. These findings (i) show that corticosteroids promote the transition from spatial to stimulus–response memory systems, (ii) provide evidence that the mineralocorticoid receptor underlies this corticosteroid-mediated switch, and (iii) suggest that a stress-induced switch from hippocampus-based to nucleus caudate-based memory systems can rescue performance.

Corticosterone, brain mineralocorticoid receptors (MRS) and the activity of the hypothalamic-pituitary-adrenal (hpa) axis: The Lewis rat as an example of increased central MR capacity and a hyporesponsive HPA axis

In this study we report a series of differences in brain and peripheral elements regulating the hypothalamic-pituitary-adrenal (HPA) axis between male LEW and Wistar rats. We found: (i) differential properties of mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) in the brain (hippocampus, hypothalamus) and pituitary: LEW rats displayed an increased capacity of MRs in the hippocampus and hypothalamus and a decreased capacity of glucocorticoid receptors GRs in the pituitary. The binding affinity (Kd) for MRs and GRs in the hippocampus was comparable. (ii) Lower concentrations of corticotropin releasing hormone (CRH) mRNA were detected in the nucleus paraventricularis of the hypothalamus of LEW rats. (iii) Adrenal weight was similar in LEW and Wistar rats; however, LEW rats had about 30% less adrenocortical cells. Subjecting adrenocortical cells to increasing doses of ACTH1-24 in vitro resulted in about a 60% smaller release of corticosterone in LEW rats. (iv) LEW rats escaped dexamethasone suppression showing increased basal levels of endogenous ACTH, but responded with a comparable release of corticosterone to the IV injection of 5 ng ACTH1-24. (v) LEW rats responded to a variety of stimuli: adrenalectomy under ether anaesthesia, a novel environment, a tail nick and restraint or an immunological challenge, with lower circulating ACTH and corticosterone plasma levels than Wistar rats. (vi) Evening levels of ACTH and corticosterone were lower in LEW than Wistar rats but did not differ in the morning. Blockade of brain MRs in the evening by a central injection of the specific MR antagonist RU28318 in LEW rats resulted in increased circulating levels of ACTH and corticosterone. (vii) Levels of corticosteroid-binding proteins were lower in one-day adrenalectomized LEW rats, indicating higher levels of free corticosterone. (viii) LEW rats had a smaller thymus than Wistar rats. Taken together, the receptor binding data correspond to a decreased neuroendocrine responsiveness of LEW rats to stress. We suggest that the shift in the central MR/GR balance of LEW rats, i.e. augmented MR-mediated effects of corticosterone, is the central regulating mechanism of the hyporeactive HPA axis in this rat strain. Lower levels of CRH mRNA in the hypothalamus and lower levels of ACTH and corticosterone in response to various stimuli, as well as the hyporesponsive adrenals to exogenous ACTH, are apparently the consequences of the life-long suppressive action of corticosterone via central MRs.

Facilitation of Feedback Inhibition Through Blockade of Glucocorticoid Receptors in the Hippocampus

In the present study the effects of intracerebroventricular (icv) and intrahippocampal administration of corticosteroid antagonists on basal hypothalamic-pituitary-adrenal (HPA) activity around the diurnal peak were compared in male Wistar rats. In two separate experiments the glucocorticoid receptor (GR) antagonist RU 38486 and the mineralocorticoid receptor (MR) antagonist RU 28318 were tested. One hour after GR antagonist injection, significant increases in plasma ACTH and corticosterone levels were observed in the icv treated rats, when compared to vehicle. In contrast, a significant decrease in ACTH levels, and a slight, but non-significant decrease in corticosterone concentrations were attained one hour after intrahippocampal injection of the GR antagonist. Injection of the MR antagonist, on the other hand, resulted in enhanced ACTH and corticosterone levels irrespective of the site of injection. These findings suggest that negative feedback inhibition at the circadian peak involves hippocampal MRs and extrahippocampal (hypothalamic) GRs. The latter feedback inhibition overrides a positive feedback influence exerted by endogenous corticosteroids through hippocampal GRs.

Spatial Learning Deficits in Mice with a Targeted Glucocorticoid Receptor Gene Disruption

Previous studies in rats using the Morris water maze suggested that the processing of spatial information is modulated by corticosteroid hormones through mineralocorticoid and glucocorticoid receptors in the hippocampus. Mineralocorticoid receptors appear to be involved in the modulation of explorative behaviour, while additional activation of glucocorticoid receptors facilitates the storage of information. In the present study we used the water maze task to examine spatial learning and memory in mice homozygous and heterozygous for a targeted disruption of the glucocorticoid receptor gene. Compared with wild‐type controls, homozygous and heterozygous mice were impaired in the processing of spatial but not visual information. Homozygous mutants performed variably during training, without specific platform‐directed search strategies. The spatial learning disability was partly compensated for by increased motor activity. The deficits were indicative of a dysfunction of glucocorticoid receptors as well as of mineralocorticoid receptors. Although the heterozygous mice performed similarly to wild‐type mice with respect to latency to find the platform, their strategy was more similar to that of the homozygous mice. Glucocorticoid receptor‐related long‐term spatial memory was impaired. The increased behavioural reactivity of the heterozygous mice in the open field points to a more prominent mineralocorticoid receptor‐mediated function. The findings indicate that (i) the glucocorticoid receptor is of critical importance for the control of spatial behavioural functions, and (ii) mineralocorticoid receptor‐mediated effects on this behaviour require interaction with functional glucocorticoid receptors. Until the development of site‐specific, inducible glucocorticoid receptor mutants, glucocorticoid receptor‐knockout mice present the only animal model for the study of corticosteroid‐mediated effects in the complete absence of a functional receptor.