Rat pup isolation distress and the brain benzodiazepine receptor.

Pharmacologic studies have demonstrated that benzodiazepines can modulate the ultrasonic vocalizations (USV) associated with social separation of rat pups. In this study, in vivo receptor autoradiography was used to determine if brain benzodiazepine receptors were functionally less available to bind an exogenous ligand during social separation. The labeled benzodiazepine receptor antagonist. 3H-RO 15-1788, was given to 10-day-old rat pups with varying schedules of social separation. In initial studies with homogenized and solubilized tissue, we found a 30% reduction in binding to cortex when pups were separated for 25 min beginning 5 min prior to tracer injection. In subsequent autoradiographic studies with this same separation schedule, the binding of 3H-RO 15-1788 was examined in 21 brain regions. Again binding was decreased in neocortex (frontal, motor, and somatosensory). In addition, we found significantly decreased binding in hippocampus, dentate gyrus, and superior and inferior colliculi. These same regions showed no alteration of in vitro binding of 3H-RO 15-1788. Therefore, these decreases in in vivo binding do not reflect changes in receptor number. The interpretation of decreased in vivo binding and implications of these results for defining the neural substrates of separation behavior are discussed.

Behavioral and autonomic responses to peer separation in pigtail macaque monkey infants.

Brief maternal separations of young nonhuman primates have been used extensively to study the behavior and physiology of attachment, loss, and bereavement. The physiological responses to the loss of alternative attachment figures, such as peers, is less well documented in nonhuman primates. This study examined both autonomic and behavioral responses of peer-reared pigtail macaque infants to separation. Eight infants were removed from their mothers at birth and reared in four peer pairs. At 6 months of age, each monkey was implanted with a multichannel biotelemetry device which transmitted heartrate, body temperature, EEG, EMG, and EOG. Blood was collected twice weekly for immunological assessment. Behavioral and physiological data, including sleep, were collected for 1 week of baseline, 2 weeks of separation, and 1 week of reunion. Behavioral and physiological results indicated agitation but not depression following separation from their peer attachment figures. We found reduced mitogenic responses to pokeweed consequent to peer separation, suggestive of altered B-cell function. REM variables were the only sleep measures affected by the separation, and were suggestive of agitation but not depression.

Cardiovascular structural changes induced by isolation-stress hypertension in the rat.

The cardiovascular structural remodelling associated with psychogenic hypertension was investigated in genetically normotensive rats subjected to isolation stress. Male Wistar rats were stressed by intermittent social isolation and compared to control rats living in groups. The stressed rats had higher systolic blood pressures than the control rats throughout the study. After 1 week of isolation, ornithine decarboxylase activity, a marker for hypertrophy, was increased in the right ventricle of the stressed rats. After 6 weeks of intermittent isolation, the myocardium of the stressed rats was hypertrophied, involving both right and left ventricles. The aorta was also hypertrophied, whereas the tail artery remained unaffected. Later, after 12 weeks of isolation, the left ventricular hypertrophy persisted whereas the right ventricle and aorta returned to normal. It seems, therefore, that social stress hypertension is accompanied by very early structural changes, which affect at least the heart and the aorta, and cannot be directly linked to the severity or duration of hypertension.

Time-dependent enhancement of lymphocyte activation by mitogens after exposure to isolation or water scheduling.

The effects of isolation and water scheduling on mitogen induced lymphocyte proliferation were investigated. Isolated rats were animals which had been raised in group-housed conditions and then transferred to individual cages with ad lib access to water for a 1 or 2 week period. Water scheduled rats were maintained in group housing (5 rats per cage) with ad lib access to food but with access to water for a single 30 minute session each day. Responses of these groups were compared to those of animals which had been continuously group-housed with ad lib access to food and water. No differences in lymphocyte responses to phytohemagglutinin (PHA) were found 1 week after exposure to isolation. However, after 2 weeks, splenic and blood T lymphocytes from isolated animals demonstrated an increased proliferative response to suboptimum and maximum concentrations of PHA. Splenic B lymphocyte responses to lipopolysaccharide (LPS) from isolated animals were also increased by 2- to 3-fold compared to group-housed controls. Two weeks of exposure of animals to daily water scheduling similarly increased the splenic lymphocyte proliferation. This increased responsiveness to PHA was not accompanied by a significant change in the sensitivity of the lymphocytes to PHA, in the total number of white blood cells, or the proportion of splenic T or T helper lymphocytes. Our results show that the increase in lymphocyte proliferation is time-dependent, requires greater than 1 week of exposure to isolation and is due to factors other than changes in sensitivity to mitogen or T lymphocyte number.

The ACTH4-9 analog ORG 2766 'normalizes' the changes in motor activities of rats elicited by housing and test conditions.

Motor activities of rats were decreased by short-term (7 days) social isolation as well as by intense light test conditions. The ACTH4-9 analog ORG 2766, s.c. administered 50 min before testing, dose-dependently decreased the high motor activities of group-housed rats tested under low light conditions and increased the low motor activities of short-term isolated rats tested under intense light conditions (ED50: 0.01-0.03 microgram/kg). Structure-activity studies suggest that the essential structure for these effects may be located in the C-terminal tripeptide Phe-D-Lys-Phe. Treatment with ACTH4-10 (100 micrograms/kg) tended to enhance some of the effects of the environmental conditions. Pretreatment of rats with the opioid antagonist naltrexone (450 micrograms/kg, s.c.) completely blocked the 'normalizing' effects of ORG 2766, implicating endogenous opioids in this action of ORG 2766. Since social behaviors of rats are similarly affected by ORG 2766 as motor activities, it is suggested that this peptide affects the integration of sensoric stimuli rather than the specific motor output systems of these behaviors.

Causes of changes in brain noradrenaline systems and later effects on responses to social stressors in rhesus monkeys: the cascade hypothesis.

Disruption of social attachments in social primates produces a protest-despair response. In rhesus monkeys, the response is probably adaptive in the feral environment, although the despair stage resembles human depression in many respects. The severity of the response varies between individuals and is affected by deprivation of certain classes of social stimuli during development. Social deprivation is associated with differences in the concentrations of noradrenaline (NA) in cerebrospinal fluid and in responses to agents that affect catecholamine systems. Thus, early rearing conditions and pre-existing genetic or perinatal differences between monkeys can have long-term effects on the response to social separation, and NA system release and/or receptor mechanisms are involved. NA systems appear to mediate adaptation to the environment from the level of perception to reorganization of neural tissue. Adaptation to the social environment may involve a cascade of changes that begins with behavioural coping attempts and terminates in structural reorganization of regions of the cerebral cortex. Processes at each level occur within environmentally appropriate but neurobiologically constrained time-frames. The cerebral NA system may be an adaptive mechanism that can fail or be damaged. Behavioural changes caused by such damage or failure would be manifested by inappropriate responses to environmental contingencies and inability to change behaviour to adapt to the prevailing environment. These features of NA system disorder could be common to depression and several other forms of human psychopathology.

Rapid effects of environmental disturbance on rat plasma unesterified fatty acid and tryptophan concentrations and their prevention by antilopolytic drugs.

1 Changes of plasma unesterified fatty acid (UFA) and tryptophan concentration in group-housed rats following removal of their cage-mates and the effects of antilipolytic drugs on these changes were investigated. 2 Removal of group-housed 24 h fasted rats but not fed rats from cages resulted in increased plasma UFA concentration in the remaining rats which was associated with significant increases of the proportion of free tryptophan but significant falls of total tryptophan concentration. These rapid changes were not associated with brain tryptophan changes. Plasma tyrosine concentration was unaffected. 3 The fall of plasma tryptophan did not appear to be due to passage into red cells as erythrocyte tryptophan concentration remained unchanged. 4 Plasma UFA concentrations correlated positively and significantly with corticosterone concentrations which were also increased following removal of cage-mates. 5 Plasma UFA increases and tryptophan changes in the fasting rats were both prevented by nicotinic acid or propranolol. Corticosterone concentration was increased by nicotinic acid but unaffected by propranolol. 6 The possible importance of these rapid changes of plasma tryptophan and of their prevention by antilipolytic drugs is discussed.

Supersensitivity to isoprenaline induced in rats by prolonged isolation.

1 The effect of isoprenaline on diastolic blood pressure and heart rate was determined in anaesthetized male rats which had been housed individually for 6-8 weeks after weaning, and compared with its effect in group-housed litter-mate controls.2 Isoprenaline caused a significantly greater fall in diastolic pressure in isolated rats and a greater increase in heart rate.3 Low doses of noradrenaline (5-10 ng) caused vasodepressor responses in isolated, but not in group-housed rats.4 The pressor response to noradrenaline, which was smaller in isolated rats, was increased to the level of group-housed controls by (+/-)-propranolol.5 Prolonged isolation of rats may bring about an increase in sensitivity of beta-adrenoceptors to isoprenaline and noradrenaline.