Alain Sarrieau

Neonatal Handling Alters Adrenocortical Negative Feedback Sensitivity and Hippocampal Type II Glucocorticoid Receptor Binding in the Rat

Adult rats handled (H) daily for the first 3 weeks of life show a dramatically altered adrenocortical response to stress. We found that H animals secreted less ACTH and corticosterone (B) during and following the termination of stress than did nonhandled (NH) controls. In contrast, H and NH animals did not differ in basal B secretion at any point in the diurnal cycle, nor in adrenocortical responses to exogenously administered oCRF or ACTH. Moreover, the clearance rate for B was similar in H and NH animals. H animals were more sensitive than NH animals to the inhibitory effects of either B or dexamethasone on stress-induced adrenocortical activity. In a dose-response study, both glucocorticoids administered 3 h prior to testing suppressed the adrenocortical response to a 20-min restraint stress to a greater extent in the H animals. Handling increased type II, glucocorticoid receptor binding capacity in the hippocampus of adult animals (approximately 50% increase in capacity, with no change in affinity). There were no handling-induced changes in type II receptor binding capacity in the hypothalamus or pituitary, nor in type I receptor binding capacity in the hippocampus. Following chronic (5 mg/kg/day) treatment with B, hippocampal type II receptor binding capacity was significantly reduced in the B-treated H animals, compared with saline-treated H animals, and indistinguishable from saline-treated NH animals. Down-regulated H animals, like NH animals, hypersecreted B following the termination of stress in comparison to the saline-treated H animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Postnatal development and environmental regulation of hippocampal glucocorticoid and mineralocorticoid receptors.

We report here on developmental changes in [3H]RU 28362 (glucocorticoid receptor) and [3H]aldosterone (mineralocorticoid receptor) binding capacity in soluble fractions prepared from hippocampal tissue. [3H]RU 28362 binding was low on Day 3 of life (less than 30% of that observed in adults) and increased towards adult values during the second and third weeks of life, a pattern virtually identical to that previously reported for dexamethasone binding. In contrast, [3H]aldosterone binding on Day 3 of life was only slightly lower than that observed in adults and reached adult values by Day 7. Postnatal handling of rat pups, which has been shown to increase dexamethasone binding in hippocampus, resulted in a significant increase in [3H]RU 28362 binding capacity in hippocampus, but had no effect on [3H]aldosterone binding.

Differential effects of social stress on central serotonergic activity and emotional reactivity in Lewis and spontaneously hypertensive rats.

Social stress by repeated defeat has been shown to be endowed with neuroendocrine and behavioural effects that render this stress model useful to identify adaptive mechanisms. Among these mechanisms, those related to central serotonergic systems (e.g., hippocampal 5-HT1A receptors, cortical 5-HT2A receptors) have been particularly underlined. Nonetheless, how (i) the neuroendocrine and behavioural effects of social stress are affected by the genetic status of the animal, and (ii) this status affects the relationships between central serotonergic systems and adaptive processes has not been studied so far. The present study has thus analysed the effects of repeated defeat (once a day for seven days) by Long-Evans resident rats upon the psychoneuroendocrine profile of Lewis rats and spontaneously hypertensive rats previously characterized for their contrasting social and anxiety-related behaviours. Repeated defeat decreased in a time-dependent manner, body weight growth and food intake in both strains, these decreases being, however, more severe and longer lasting in Lewis rats. This strain-dependent difference could not be accounted for by differences in physical contacts with the resident rats as the number of attacks and their latency throughout the stress period were similar between spontaneously hypertensive and Lewis rats. When exposed to an elevated plus-maze test of anxiety, the unstressed Lewis rats entered less the open arms than their spontaneously hypertensive counterparts, thus confirming that Lewis rats are more anxious than spontaneously hypertensive rats. This difference was amplified by stress as the latter increased anxiety-related behaviours in Lewis rats only. These strain- and stress-related differences were associated with differences in locomotor activity, this being increased in unstressed Lewis compared with spontaneously hypertensive rats; moreover, stress triggered hypolocomotion in the former but not the latter strain. Lastly, in the forced swimming test. Lewis rats spent more time immobile than spontaneously hypertensive rats with stress increasing immobility in a strain-independent manner. Beside the aforementioned metabolic changes, the activity of the hypothalamo-pituitary-adrenal axis was slightly stimulated in a strain-independent manner by the stressor, as assessed by increased corticosterone levels and adrenal weights, and decreased thymus weights. In Lewis, but not in spontaneously hypertensive rats, midbrain serotonin metabolism was increased by stress, a difference associated with an increased Bmax value of cortical [3H]ketanserin binding at 5-HT2A receptors. On the other hand, the Bmax value of hippocampal [3H]8-hydroxy-2-(di-n-propylamino)tetralin binding at 5-HT1A receptors was decreased by stress, this reduction being amplified in spontaneously hypertensive compared with Lewis rats. This study shows that the psychoneuroendocrine responses to social stress may have a genetic origin, and that the use of socially stressed Lewis and spontaneously hypertensive rats may provide an important paradigm to study adaptive processes. However, whether the aforementioned strain-dependent differences in central serotonergic systems (partly or totally) underlie the distinct profiles of emotivity measured in spontaneously hypertensive and Lewis rats, is discussed in the context of the relationships between serotonergic systems and behavioural responses to novel environments.

Behavior and pituitary-adrenal function in large white and Meishan pigs.

Six-wk-old piglets of both sexes from European Large White (LW, n = 36) and Chinese Meishan (MS, n = 24) breeds were individually exposed to a novel environment, a stressful stimulation. Behavioral and pituitary-adrenal reactivity were investigated. When compared with LW, MS pigs displayed low locomotion (18.5 +/- 2.2 vs. 41.0 +/- 3.8 squares crossed/10 min; P < 0.0001), and defecation scores (0.58 +/- 0.15 vs. 4.86 +/- 0.37 fecal boli; P < 0.0001). Basal concentrations of cortisol were higher in MS (96.1 +/- 1.1 vs. 44.9 +/- 1.1 ng/ml; P < 0.0001), although no differences between breeds were found in basal concentrations of adrenocorticotropic hormone (ACTH). In response to novel environment exposure, the ACTH increase was greater in LW than in MS, but the cortisol response was not different on a log scale. To further investigate the pituitary-adrenal differences between the two breeds, the 24-hr profile of ACTH and cortisol plasma concentrations, a corticotropin-releasing factor (CRF) and a coupled dexamethasone-ACTH test were studied. Five castrated male 9-wk-old piglets from each breed were fitted surgically with a jugular vein catheter. A classic marked circadian rhythm of cortisol and a weak nycthemeral variation of ACTH were found. Cortisol concentrations were approximately twice higher in MS exclusively during the early light phase (from 0800-1200 hr) of the cycle, but no significant interbreed difference was found in the circadian rhythm of ACTH. Administration of CRF (1 microgram/kg iv) induced the same significant increase in plasma ACTH and cortisol concentrations in both breeds. Administration of ACTH (10 micrograms/kg i.v.) increased significantly cortisol concentrations and revealed no difference in plasma cortisol response to ACTH. These data suggest that the hypercortisolism of MS pigs is of adrenal origin, and related to extrapituitary factors that control the adrenal sensitivity during the light phase of the diurnal cycle.

Characterization and autoradiographic distribution of vasoactive intestinal peptide binding sites in the rat central nervous system.

Biochemical characteristics and topographical distribution of mono-[125I )vasoactive intestinal peptide (VIP) binding sites in rat brain were studied on tissue sections and by quantitative autoradiography. Biochemical investigations show two classes of binding sites with a dissociation constant of 1.03 +/- 0.11 nM and 68 +/- 14 nM and a maximal binding capacity of 43.3 +/- 5.1 fmol/mg protein and 713 +/- 117 fmol/mg protein respectively. The order of potency of various peptides to inhibit 125I-VIP binding to brain sections is: VIP greater than PHI greater than secretin greater than VIP greater than hGRF. Autoradiography reveals the highest densities of binding sites in the pineal gland, the dentate gyrus of the hippocampus, the central amygdaloid nucleus and in various thalamic nuclei such as the mediodorsal, lateral posterior, submedius, dorsolateral and medial geniculate nuclei. Similar high densities are observed in the olfactory bulbs as well as in the suprachiasmatic and dorsomedial nuclei of the hypothalamus and in the superior colliculus. These data together with the distribution of the endogenous peptide suggest a physiological role for VIP both in the regulation of CNS activities and pituitary functions.

Characterization and autoradiographic distribution of neurotensin binding sites in the human brain

The characteristics and topographical distribution of monoiodo 125I-Tyr3-neurotensin (NT) binding sites in normal human brain tissue were studied on brain sections and by quantitative autoradiography. Sections at the level of the substantia nigra show a dissociation constant and maximal binding capacity of 4.8 +/- 0.8 nM and 70 +/- 7 fmol/mg protein, respectively. High density of 125I-NT binding sites were mainly found in dopaminergic (DA)-rich areas such as the substantia nigra, the ventral tegmental area, the striatum and the nucleus accumbens, further supporting an interaction between NT and DA neurons in human brain.

CONTRIBUTION OF SEX AND GENETICS TO NEUROENDOCRINE ADAPTATION TO STRESS IN MICE

Male and female C57BL/6 (B6) and DBA/2 (D2) mice were subjected to either acute or 5 days of repeated restraint in ventilated, 50 ml centrifuge tubes. Control animals were not disturbed. The acute restraint animals were killed immediately following 15, 30 or 60 min of restraint and blood collected for corticosterone (CORT) analysis. The results of the acute restraint procedure revealed a strain difference in time to peak CORT in plasma with D2 animals showing an earlier peak. The males of both strains evinced similar maximum response and similar to B6 females; however, the D2 females showed a 2-fold greater CORT response than did the B6 females. Repeated restraint consisted of 5 days of 12 h in the tubes. At the end of 5 days, the animals were weighted and adrenalectomized in preparation for determination of brain corticosteroid receptors. Upon sacrifice, brains, thymus, adrenals and blood were harvested, the last for corticosteroid binding globulin (CBG). Five days of repeated restraint produced body weight loss in both strains, with B6s less affected than D2s. Repeated restraint reduced the mass of the adrenals in the B6s only. Restraint also reduced the mass of the thymus in both strains and sexes, but to a greater extent in the B6s. Plasma CBG densities were also sensitive to restraint, but only in females, showing a restraint-related decrease. Repeated restraint had no effect on hippocampal glucocorticoid or mineralocorticoid receptors; however for the latter, we observed significant strain and sex effects with D2 having higher Bmax than B6 and females having higher Bmax than males. In the pituitary, glucocorticoid receptors (GR) were reduced by repeated restraint in males, but increased in females, especially in the B6. These findings lend preliminary evidence for involvement of sex and genetics as sources of individual differences in bioadaptation to stress.

Autoradiographic localization of glucocorticosteroid and progesterone binding sites in the human post-mortem brain

The presence of glucocorticosteroid and progesterone binding sites in the human brain has been investigated with an in vitro autoradiographic approach using [3H]RU 38486. The present data revealed that the hippocampus, the entorhinal cortex, the subiculum, the fimbria and the amygdala showed a different pattern of [3H]RU 38486 labeling using various unlabeled steroids (RU 38486, RU 28362, cortisol, RU 27987 and dexamethasone). RU 38486 is the best competitor in all these structures. The hippocampus seems to possess mainly glucocorticosteroid binding sites whereas the entorhinal cortex and the subiculum reveal the presence of both glucocorticosteroid and progesterone binding sites. Furthermore, the amygdaloid complex and the fimbria show a high density of glucocorticosteroid binding sites.

Behavioral and neuroendocrine reactivity to stress in the WKHA/WKY inbred rat strains: a multifactorial and genetic analysis

Genetic factors have been shown to influence the nature and the intensity of the stress responses. In order to understand better the genetic mechanisms involved, we have studied the behavioral and neuroendocrine responses to novel environments in the WKHA/WKY inbred strains and we have investigated the genetic relationships between these traits in a segregating F2 intercross. The animals were submitted to behavioral tests known to provide both indices of activity and fear (activity cages, open field and elevated plus-maze). The plasma levels of prolactin, ACTH, corticosterone, glucose and renin activity were determined after a 10-min exposure to novelty. Our results showed that WKHA rats, compared to WKYs, were more active in a familiar as well as in novel environments. They exhibited also less anxiety-related behaviors and lower neuroendocrine responses. A principal component analysis performed on the behavioral F2 results defined three independent factors: general activity, anxiety and defecation, none of them being correlated with the neuroendocrine measures. Thus this study suggests that these different responses to stress are independent components that may have distinct molecular bases.

Regulation of interleukin-1 receptor expression in mouse brain and pituitary by lipopolysaccharide and glucocorticoids.

Interleukin-1 (IL-1) receptors have been characterized in mouse pituitary and brain. Previous studies have demonstrated that IL-1 receptor density is high in the dentate gyrus in the hippocampus and that lipopolysaccharide (LPS) injection caused an 80% decrease in the number of hippocampal IL-1 receptors, while pituitary receptors in the anterior pituitary were unaffected. In order to investigate on the role of glucocorticoids (GC) in the control of IL-1 receptor expression in the brain and pituitary, the effect of stress, exogenous GC administration or adrenalectomy (ADX) on IL-1 receptor density was determined. Assays were achieved under basal and LPS-stimulated conditions by in situ quantitative autoradiography technique using human recombinant 125I-IL-1 alpha as a tracer. An increase of GC concentration in serum, following immobilization stress or dexamethasone (DEX) treatment (short and long term), did not modify, in the hippocampus, the density of IL-1 receptors under basal conditions or after peripheral LPS injection. On the contrary, ADX significantly decreased IL-1 receptor density in LPS-treated animals. In the anterior pituitary, a significant increase in the density of basal IL-1 receptors was observed 6 h following immobilization stress or after 7 days of DEX treatment while short-term DEX treatments are ineffective. In contrast to what was observed in the hippocampus, no changes in pituitary receptor densities were observed in ADX mice. These results indicate that hippocampal and pituitary IL-1 receptor expressions are differentially regulated by GC. Therefore, this report constitutes the first demonstration of an in vivo regulation of IL-2 receptors in the pituitary.