Biochemical indices of reactivity and habituation in rats with hippocampal lesions

The response of rats with hippocampal lesions to acute and repeated footshock stress was assessed by measurement of pituitary cyclic AMP, plasma corticosterone and plasma prolactin. Levels of pituitary cyclic AMP and plasma prolactin and corticosterone were similar in never-shocked sham controls, and never-shocked hippocampal and neocortical lesion groups. Acute first time shock markedly elevated all measured stress indices with no statistically significant differences observed among surgical groups. In rats subjected to repeated stress (one 15 min footshock session per day for 10 days) and sacrificed 24 hours after the last shock session, levels of pituitary cyclic AMP and plasma hormones were similar to levels in never-shocked shams with the exception of the hippocampal animals. The rats with hippocampal lesions had higher levels of pituitary cyclic AMP, plasma corticosterone and plasma prolactin compared to never-shocked animals. We suggest that these data reflect a hyperreactive response of the hippocampal animals to a situation previously associated with shock. Finally, rats in all surgical groups subjected to repeated stress and sacrificed immediately after the last shock session showed a diminished cyclic AMP response to the stressor as compared to first footshock session response, demonstrating a habituation to the stressor as measured by this index. No differences in habituation were observed among hippocampal, neocortical and sham groups. Plasma hormone responses did not habituate in any group. These data support the behavioral observations of hyperreactivity in hippocampal animals and indicate that hippocampal animals are able to habituate to repeated stressful stimuli.     

Cyclic AMP and cyclic GMP response to stress in brain and pituitary: Stress elevates pituitary cyclic AMP

Male rats were exposed to six stressors (saline injection, cold, forced running, Formalin injection, immobilization, electric footshock) for 15, 30, or 60 min. Following sacrifice by microwave irradiation, cyclic AMP and cyclic GMP levels were measured in pituitary, pineal and 8 regions of rat brain. All stressors except saline increased plasma corticosterone, plasma prolactin and pituitary cyclic AMP levels compared to control animals. The magnitude of the pituitary cyclic AMP response was highly correlated with the intensity of the stress as determined by the levels of plasma prolactin. Electric footshock increased pituitary cyclic AMP levels over 10 fold and plasma prolactin over 60 fold. Cyclic AMP levels in other brain regions were not altered. Cerebellar cyclic GMP was increased only by stressors that involved increased motor activity.     

Influence of repeated cocaine exposure on the endocrine and behavioral responses to stress in rats

Previous studies have determined that chronic cocaine exposure inhibits the serotonergic stimulation of hormone secretion. The present experiments were conducted to determine whether the endocrine responses to stress could be a useful approach to assess the influence of cocaine exposure on neuronal function. Male rats received twice daily injections of cocaine (1–15 mg/kg, IP) for 7 days. Animals were subsequently exposed to different stressors, i.e. conditioned emotional stress utilizing a low (0.5 mA) or high (1.5 mA) intensity footshock during training, or to immobilization stress. Immediately after the stress procedures, blood samples were collected for radioimmunoassay of plasma corticosterone, prolactin, and renin concentrations. Repeated cocaine exposure attenuated the stress-induced elevations of corticosterone and prolactin secretion, and attenuated some of the behavioral effects of the low intensity conditioned emotional stress. When exposed to the high intensity conditioned emotional stress, cocaine did not alter the endocrine or behavioral effects of stress. Finally, repeated cocaine exposure modified the immobilization stress-induced elevation of renin secretion; low doses of cocaine (1 or 5 mg/kg) attenuated, while higher doses (10 mg/kg) potentiated the renin response to immobilization stress. Thus, the influence of repeated cocaine exposure on the endocrine and behavioral responses to stress appears to depend upon the type and intensity of the stressor. Compared with previous studies which found altered neuroendocrine responses to serotonin releasers and agonists following cocaine exposure, the hormonal responses to stress are less consistently modified by cocaine.     

Single and repeated air blast stress and brain histamine

Exposure of rats to air blasts for 1, 5 and 15 min resulted in a significant increase in plasma corticosterone level and in the hypothalamic histamine concentration. Midbrain histamine content was increased after 1 and 5 min of exposure but cortical histamine increased following 1 min of exposure only. Stress of longer duration (30 mins did not significantly affect histamine concentration in any of the three brain regions investigated, although plasma corticosterone level remained very significantly (14.5-fold) elevated. Repeated exposure of rats to air blasts of 15 min duration resulted in a significant elevation of hypothalamic histamine concentration while midbrain and cortical histamine was not significantly altered. Plasma corticosterone level was again very significantly (10-fold) increased. Present results suggest the involvement of brain histamine in the response to stress.     

Effects of naloxone on corticosterone response to stress

The effect of naloxone on the corticosterone response to restraint stress was examined. Naloxone (8 mg/kg, IP) did not alter basal corticosterone or the magnitude of the corticosterone response to restraint stress. Naloxone did, however, retard the fall in corticosterone following the end of restraint stress; thus the drug prolonged the stress response. These data suggest that endogenous opiates play a role in the restoration of corticosterone levels back to normal after stress.     

Chronic unpredictable stress, but not chronic predictable stress, enhances the sensitivity to the behavioral effects of cocaine in rats

RATIONALE: Chronic unpredictable stress, in which the type and timing of stress exposures are varied, alters protein levels in the mesolimbic DA system in a manner previously shown to be associated with enhanced behavioral responsiveness to cocaine. Chronic exposure to the same or predictable stress (restraint) does not. Thus, we examined the effects of chronic unpredictable and chronic predictable (restraint) stress on the locomotor activating and place conditioning effects to low cocaine doses. OBJECTIVE: To test whether chronic unpredictable stress enhances the sensitivity to the behavioral effects of cocaine. METHODS: Rats were exposed to 10 days of chronic unpredictable stress, of chronic predictable (restraint) stress, or were not stressed. One day following cessation of stress exposure, locomotor activity to cocaine (0 or 7.5 mg/kg) was assessed for 4 consecutive days and corticosterone levels on the last day were determined. In other experiments, the effects of the chronic stress procedures on cocaine (0.5 and 7.5 mg/kg) place conditioning using an unbiased procedure were assessed. RESULTS: Chronic unpredictable, but not chronic predictable, stress transiently increased the locomotor activating effects of cocaine and this was correlated positively with corticosterone levels. Chronic unpredictable, but not chronic predictable, stress also enhanced the place conditioning effects of cocaine: increased place preference was seen with the low dose and a pronounced place aversion occurred with the high dose. CONCLUSIONS: These data demonstrate that chronic unpredictable stress enhances the behavioral effects of cocaine, including its aversive effects, whereas chronic predictable stress (restraint) is without effect.     

Prior stress attenuates the analgesic response but sensitizes the corticosterone and cortical dopamine responses to stress 10 days later

This study demonstrates that pre-exposure to stress influences subsequent effects of stress on pain sensitivity (stress-induced analgesia) and on plasma corticosterone and brain catecholamine activity. Animals exposed to a 30 min shock session (S1=8, 5.0 s shocks) 10 days earlier showed a significant attenuation of shock-induced analgesia, as measured by increased latency of tail withdrawal from a hot water bath immediately after a 40 s, 1.6 mA footshock (S2). Animals exposed to shock 10 days before testing also exhibited a higher plasma corticosterone response to testing than did all other groups. Norepinephrine (NE) levels in the frontal cortex and dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) levels in the frontal cortex and nucleus accumbens were not altered in any group. However, the DOPAC/DA ratio in the frontal cortex was increased by analgesia testing, and this increase was enhanced only by the combination of shock 10 days before testing and shock immediately before the test (S1+S2). These results are consistent with previous reports from this laboratory which indicate that an animal's acute response to stress is strongly influenced by its past history of stress.     

Comparison of the antidepressant sertraline on differential depression-like behaviors elicited by restraint stress and repeated corticosterone administration

Depressive disorder involves emotional, cognitive, autonomic and endocrine alterations and also evidences support the role of stress in the development of this disorder. Because the hypothalamic-pituitary-adrenal axis is involved in the stress response with a concomitant rise in plasma corticoids, the present study compares the antidepressant effects of sertraline (10 mg/kg, i.p.) on behavioral changes elicited by (i) restraint stress (2.5 h/day for 13 days) and (ii) corticosterone injections (30 mg/kg, s.c., for 13 days). Stressed animals, but not corticosterone-treated animals displayed anxiety behavior and a reduction in the acquisition of a conditioned avoidance response to 25% of control levels (8.0 ± 2.2 vs. 31.7 ± 3.2), being this effect partly sensitive to sertraline. Stressed, but not corticosterone-treated, animals displayed an increased escape failure compared with the control group (24.6% ± 3.5 vs. 1.6 ± 0.7), an effect partly prevented by sertraline treatment (7.3% ± 2.0). Both stressed rats and corticosterone-treated rats showed an increase in immobility in the forced swim test, an effect prevented by sertraline. These results suggest that the altered behaviors elicited by stress and corticosterone can be explained by neural modifications that are sensitive to the sertraline antidepressant.     

Exposure of the amygdala to elevated levels of corticosterone alters colonic motility in response to acute psychological stress

The amygdala is important for integrating the emotional, endocrine and autonomic responses to stress. Exposure of the amygdala to elevated levels of corticosterone (CORT) induces anxiety-like behavior and a hypersensitive colon in rodents; however, effects on colonic transit are unknown. Micropellets releasing CORT alone or combined with a selective glucocorticoid (GR) or mineralocorticoid (MR) receptor antagonist were implanted bilaterally at the dorsal boundary of the central amygdala in male rats. Inactive cholesterol implants served as controls. Seven days later, rats received water avoidance stress (WAS) for 1 h and the fecal pellet output was measured. Colorectal transit was also evaluated following the stressor by recording the time for expulsion of a glass bead placed into the colorectum. Plasma CORT levels were evaluated before WAS, after 60 min of WAS and 90 min post-WAS. Exposure of the amygdala to elevated CORT did not alter daily fecal pellet production or the number of fecal pellets released in response to WAS. However, following WAS, rats with CORT implants on the amygdala showed a delay in colorectal transit compared to cholesterol-implanted controls. Plasma CORT measurements showed that basal and WAS-induced increases in plasma CORT were similar in all groups but a prolonged increase in plasma CORT was observed 90 after cessation of WAS in rats with CORT implants. The post-WAS changes in colonic motility and plasma CORT were prevented by antagonism of GR or MR in the amygdala, suggesting their importance in driving stress-associated changes in colonic motility.     

Effect of prolonged exposure to nicotine and stress on the pituitary-adrenocortical response; the possibility of cross-adaptation

Daily IP injections of nicotine (200 μg/kg body weight) resulted in an adaptation of the nicotine induced rise in plasma corticosterone. By 30 days the plasma corticosterone rise was not significantly different from that seen in control animals receiving an injection of saline. A similar adaptation to the plasma corticosterone response to the stress of signalled, irregular footshock was also observed. However, in the case of the exposure to stress, while the corticosterone response at day 40 was significantly less than the response seen on day 1, it was still significantly greater than the plasma corticosterone level from unstressed control animals. Cross-adaptation experiments were conducted in which animals were adapted to the steroidogenic action of nicotine and then subjected to a novel exposure to footshock stress, and vice versa. In both situations the animals responded to the novel stimulus, either stress or nicotine, with a significant rise in plasma corticosterone. It was postulated that nicotine and psychological stress act upon the hypothalamo-pituitary-adrenal axis via functionally separate pathways at the level of the corticotrophin releasing factor neuron. The separate pathways appear to differ in their ability to be inhibited by corticosterone feedback.     

The behavioural effects of levallorphan,cyprenorphine (M 285) and amphetamine on repeated Y-maze performance in rats

The effects of levallorphan and cyprenorphine (M 285) were studied on repeated Y-maze performance in rats. It was postulated that these narcotic antagonists—implicated as psychotomimetics—would disrupt the experiencing process and augment fear-motivated behaviour. These compounds produced differential effects on behaviour. Levallorphan induced bizarre-excitation and disrupted habituation; cyprenorphine was ineffective in these respects. No augmentation of fear-motivated behaviour was observed. Amphetamine increased locomotor activity without affecting habituation.     

Effect of stress on brain histamine

Stress of short duration (5 min) resulted in a significant increase in plasma corticosterone level and a significant decrease in the midbrain histamine concentration in rats. Exposure to 15 min stress caused a significant elevation in the hypothalamic histamine level. Stress of longer duration (30 or 60 min) did not affect hypothalamic, cortical or midbrain histamine concentration although plasma corticosterone level remained elevated. Repeated exposures of rats to 15 min stress did not significantly alter histamine concentration of any of the brain regions studied. Plasma corticosterone concentration was only 28% of that observed in animals exposed to single 15 min stress. Present data suggest a role of brain histamine in response to stress.     

Locomotor and stress responses to nicotine differ in adolescent and adult rats

Since adolescence is a critical period for the initiation of tobacco use, we have systematically compared behavioral and endocrine responses to nicotine in Sprague-Dawley rats of both sexes at early adolescence (postnatal day (P) 28), mid- adolescence (P38) and adulthood (P90). Locomotion and center time in a novel open field were evaluated for 30 min following intravenous injection of saline or nicotine (60 μg/kg), followed by measurement of plasma corticosterone. Complex age and sex differences in behavioral and endocrine response were observed, which were dependent on the functional endpoint examined. Whereas there were age differences in nicotine effects on all functional measures, sex differences were largely restricted to adult stress-related corticosterone and center-time responses. Although significant drug effects were detected at P28 and P90, there was no effect of nicotine at P38 on any measure examined. In saline-treated males, but not females, there were significant positive correlations across age between initial ambulatory counts and both initial vertical counts and total center time. Nicotine treatment increased correlations in both sexes, and yielded a significant negative interaction between initial ambulatory counts and plasma corticosterone. The unique responses of adolescents to nicotine are consistent with an immature function of nicotinic acetylcholine receptors at this age.     

Raised corticosterone in the rat after exposure to the elevated plus-maze

Rats given one or two 5-min trials in the elevated plus-maze had plasma corticosterone concentrations significantly higher than the home cage control group and there was no sign of habituation in the group given two trials. In rats given two plus-maze trials the corticosterone responses were significantly higher in the group given 10-min rather than 5-min trials. A previous experience of cat odour (1 week earlier) has no effect on the plasma corticosterone response, but did have an anxiogenic effect that could be detected by a decrease in the percentage of time spent on the open arms of the plus-maze. The results are discussed with reference to the nature of anxiety generated by trials 1 and 2 and by the trial duration in the plus-maze, and with respect to dissociation between behavioural and endocrinological measures.     

The corticosterone stress response and mercury contamination in free-living tree swallows, Tachycineta bicolor

We determined mercury concentrations in tree swallows, Tachycineta bicolor, from Massachusetts and Maine with different levels of contamination. Baseline and stress-induced plasma corticosterone concentrations from adults and nestlings (Massachusetts only) were compared with mercury concentrations. In Massachusetts, adult baseline corticosterone was negatively correlated with blood mercury, but showed a nearly-significant positive correlation with feather mercury. There was a negative relationship between baseline corticosterone and blood mercury in nestlings and between baseline corticosterone and egg mercury. There was no relationship between mercury and stress-induced corticosterone in any of the groups, or with baseline corticosterone in Maine sites where mercury levels were lower. The findings suggest blood and egg mercury may be a better indicator of current condition than feather mercury. Further, mercury contamination may not alter stress-induced corticosterone concentrations in tree swallows but appears to have a significant impact on baseline circulating corticosterone.     

Association between increase in cyclic AMP and subsequent induction of tyrosine hydroxylase in rat adrenal medulla

Summary After male rats (100 g body weight) have completed 7 min of swimming at 15°C, their rectal temperature is decreased by 15°C. As expected, the increase of cAMP and the decrease of cGMP concentrations in adrenal medulla are delayed by the time period necessary for the body temperature to return to normal. Thus, taking into consideration the delaying effect of hypothermia, the swimming stress experiments are in agreement with the view that the enhancement of cyclic AMP/cGMP concentration ratios may function as the second messengers for the induction of tyrosine hydroxylase in adrenal medulla.     

Dissociation between changes in cyclic AMP and subsequent induction of TH in the rat superior cervical ganglion and adrenal medulla

Summary Changes in the levels of cyclic AMP were studied in the superior cervical ganglion and adrenal medulla of male Sprague-Dawley rats under experimental conditions which lead to a trans-synaptic induction of tyrosine hydroxylase (TH) in these organs.In the superior cervical ganglion an intermittent 2-hour swimming stress or treatment with reserpine (5 or 10 mg/kg s.c.) failed to change cyclic AMP levels significantly (P>0.1) while 48 h later TH was markedly (P<0.001) increased. In the adrenal medulla the increase in cyclic AMP occurring during cold exposure was much greater than that observed during swimming stress. However, the increase in TH activity resulting from swimming stress was markedly greater then that resulting from cold exposure. The same inverse relationship holds true when comparing the effect of reserpine with that of cold stress. Moreover, during cold exposure cyclic AMP levels were maximal (700%) within 20 min and had fallen almost to control levels (180%) after 60 min. However, no significant increase in TH (P>0.1) was observed 48 h after a 1-hour cold stress whereas a 2-hour cold stress produced a marked (P<0.001) increase in TH activity. Thus, the second hour of cold stress during which cyclic AMP was only marginally elevated seems to be essential for initiating the biochemical events ultimately leading to an increased synthesis of TH.It is concluded that either cyclic AMP is not involved in trans-synaptic induction of TH or that changes in a small pool are essential and are overshadowed by a much larger pool of cyclic AMP not reacting in the same way.     

Long-term effects of fetal ethanol exposure on pituitary-adrenal response to stress

Pregnant female rats were fed either a 5.0–5.5% w/v ethanol-containing liquid diet ad lib or pair-fed the isocaloric control diet during gestation weeks 2 and 3. At 75–105 days of age, female offspring of the ethanol-treated dams showed significantly greater corticosterone responses than pair-fed- or normally-derived offspring to the stress of cardiac puncture or of noise and shaking, while pituitary-adrenal responses to exposure to a novel environment, cold or 2–3 days of fasting were normal. Adrenal sensitivity to ACTH in dexamethasone-suppressed adult offspring was unaffected by the prenatal treatment. The results demonstrate that fetal ethanol exposure enhances adult pituitary-adrenal responses to certain stressors, including alcohol as demonstrated previously, and suggest that the long-term effects may be mediated by developmental actions of alcohol on central neural mechanisms involved in the regulation of this neuroendocrine system.     

Sound vibration,a non-invasive stress: antagonism by diazepam

Rats, subjected to sound-vibration stress, showed an abrupt increase in plasma corticosterone (CS). This stimulation was reliably produced using a Burgess brand vibro-graver, a standard tool used for engraving. With the tool set at 8 or coarse, the barrel of the tool was placed on the animal's flank and the point held against the side of the metal cage for 15 s. Plasma CS increased to 29.3±4.7 µg/dl at 15 min and 15.7±1.8 µg/dl at 30 min. These levels were significantly higher than animals pretreated with diazepam, 5 mg/kg IV, 2 h prior to stimulation (9.2±2.0 and 7.4±1.5 µg/dl, respectively). Animals which were pretreated with CGS-8216 (a mixed agonist/antagonist at the benzodiazepine receptor), 2 mg/kg IV, 30 min prior to diazepam had the protective effects of diazepam abolished. Sound/vibration produced a significant elevation in plasma CS in animals given CGS-8216 alone; but, this elevation was significantly lower than in vehicle-treated controls. This comparatively lower plasma CS level suggests a partial-agonist, diazepam-like effect by CGS-8216. Experiments were done in conscious unrestrained male Sprague-Dawley rats with chronic IV catheters. Except for 15 s stimulation exposure, all animals remained isolated in sound-attenuated one-way vision boxes for the duration of the serial blood sampling. Control stimulation exposure involved similar handling without turning on the engraving tool. These results demonstrate: 1) the usefulness of this tool to provide a repeatable stress stimulus; 2) the ability of diazepam to abolish the stress response; 3) that CGS-8216 can antagonize the action of diazepam; and 4) a demonstration of the partial agonist effects of CGS-8216.     

Rapid and discrete changes in hypothalamic catecholamine nerve terminal systems induced by audiogenic stress, and their modulation by nicotine-relationship to neuroendocrine function

The effects of acute audiogenic stress, with or without simultaneous nicotine treatment (0.3 mg/kg i.v.), on catecholamine levels in discrete dopamine and noradrenaline nerve terminal systems of the hypothalamus, and on the secretion of adenohypophyseal hormones and of corticosterone, were studied using quantitative microfluorometric evaluations of catecholamine stores and radioimmunoassays for the determination of serum hormone levels. Audiogenic stress and nicotine induced very rapid and discrete decreases in noradrenaline levels in the subependymal layer (SEL), in the parvocellular part of nuc. paraventricularis hypothalamic (PA FP) and in the posterior periventricular hypothalamic systems, (PV II); the decreases were apparent 2 min following the onset of treatment. Increases of arterial blood pressure were observed after nicotine treatment but could not have been a major factor in producing the changes in catecholamine levels. These changes in NA levels may be related to the nicotine- and stress-induced increases of ACTH (SEL and PA FP) and prolactin secretion (PV II) found in the present experiments. Stress enhanced the rapid but variable increase in vasopressin secretion induced by nicotine, suggesting one possible mechanism by which stress combined with smoking can contribute to the development of increased arterial blood pressure and finally to sustained hypertension.