Harry N. Davis

Plasma Cortisol Levels of Dogs at a County Animal Shelter

Plasmacortisol levels were examined to assess the stress of dogs in a county animal shelter. Groups of dogs confined in the shelter for their 1st, 2nd, or 3rd day had higher cortisol levels than did a group maintained in the shelter for more than 9 days. Dogs in the shelter for an intermediate period (Day 4-9) had intermediate levels of cortisol. The cortisol concentrations of dogs during their first day in the shelter were greater than either those of the same dogs on Day 4/5 in the shelter or those of a group of pet dogs sampled in their own homes. There was no overall effect of 20 min of social interaction with a human (e.g., petting) on the plasma cortisol levels of dogs in the shelter on Day 1-3. However, the gender of the petter did affect cortisol levels. Those dogs interacting with a female had lower cortisol concentrations at the end of the session than did dogs interacting with a male. The results suggest that confinement in a public animal shelter produces a prolonged activation of the hypothalamic-pituitary-adrenal axis. Further, it appears that some subtle aspect of interaction with a human may be capable of moderating this response. Possible implications for the welfare of confined dogs, and for the development of behavior problems in dogs obtained from shelters, are discussed.

Stress During Pregnancy Alters Rat Offspring Morphology and Ultrasonic Vocalizations

Stress during pregnancy, or prenatal stress, is known to alter offspring behavior, morphology and physiology. We found that a heat, light and restraint stressor applied during the third trimester of pregnancy: 1) decreased the weight gain of adult female rats during pregnancy; 2) reduced the weight of pups, as well as the anogenital distance of male offspring, at birth; and 3) increased the number of ultrasonic vocalizations emitted by pups during isolation in a novel environment on Postnatal Day 14. These results closely approximate those we previously observed after peripheral administration of corticotropin-releasing factor to pregnant females during the third trimester. Together, the studies strongly suggest a role for corticotropin-releasing factor and/or other hormones of the hypothalamic-pituitary-adrenal system in mediating some of the effects of gestational stress.

Changes in the hormonal concentrations of pregnant rats and their fetuses following multiple exposures to a stressor during the third trimester.

Human and animal studies indicate that stress during pregnancy can exert long-term effects on the development of the offspring, effects that appear to be mediated in part by the hypothalamic-pituitary-adrenal (HPA) axis. In this experiment changes in levels of a variety of HPA and other hormones in both pregnant rats and their fetuses were investigated. Trunk blood was collected from pregnant females and fetuses following repeated 45-min presentations of restraint, bright lights, and heat during the third trimester. In addition, testes were harvested from the male fetuses. Hormone concentrations were determined by radioimmunoassay. Pregnant females had elevated titers of plasma corticosterone, aldosterone, and ACTH for approximately 15 min following termination of the stressor. No differences were found for beta-endorphin or prolactin. Fetuses showed a pattern of changes in plasma corticosterone and aldosterone that was similar to that of pregnant females, but no effect was observed for fetal ACTH titers. These results are consistent with a role of the HPA axis in the effects of gestational stress. Testicular levels of CRF on gestational day 21 were lower in fetuses of stressed females than in those of nonstressed females. The reduced levels of testicular CRF suggest that CRF may be involved in the altered pattern of sexual differentiation of males stressed during gestation.

CRF Administered to Pregnant Rats Alters Offspring Behavior and Morphology

Pregnant rats injected with 20 micrograms of corticotropin-releasing factor (CRF) from day 14 through 21 gained less weight during gestation than did saline-injected controls. The offspring of CRF-injected females differed from the offspring of control females in several ways: males and females weighed less during the first 2 weeks of life, males had shorter anogenital distances at birth, and males and females emitted more ultrasonic vocalizations during isolation in tests at 6 and 14 days of age. These effects are similar to those that have been observed following exposure of pregnant females to stressors, and provide support for the notion that CRF and/or CRF activation of the hypothalamic-pituitary-adrenal axis mediate effects of gestational stress.

Effects of centrally administered corticotropin-releasing factor (CRF) and α-helical CRF on the vocalizations of isolated guinea pig pups

Intraventricular corticotropin-releasing factor (CRF) was administered to guinea pig pups both with a freehand injection technique and via indwelling cannula. Behavioral effects depended upon the technique used. The highest dose of CRF (5 micrograms) inhibited the vocalizing of pups in a subsequent isolation test only when CRF was given by freehand injection. The possibility that disturbance attendant to the freehand procedure can account for this difference is discussed. To determine the effect of endogenous CRF in the absence of additional disturbance, the CRF antagonist alpha-helical CRF (ahCRF) was administered with the indwelling cannula procedure. ahCRF enhanced vocalizing during the first 10 min, and enhanced locomotor activity during the last 10 min, of a 30-min isolation test. Overall, the results indicate that endogenous CRF reduces vocalizing and locomotion during social isolation and that under certain injection conditions exogenous CRF can exacerbate the behavioral effect. The results also demonstrate the potential impact of the technique used to administer exogenous CRF. Further, the prevailing view, that CRF mediates stress-related behavioral responses, is supported only if behavioral inhibition, rather than vocalizing or locomotor activity, is viewed as the stress-related response in this situation.

Stress during pregnancy alters the offspring hypothalamic, pituitary, adrenal, and testicular response to isolation on the day of weaning

Subjecting pregnant female rats to situations that activate the hypothalamic-pituitary-adrenal (HPA) axis can have long-term effects on the development of the offspring. Restraint under bright lights is a common method of stressing pregnant females that results in consistent behavioral changes in the offspring. We investigated the effects of gestationally administered restraint, bright lights, and heat on the HPA axis response of 21-day-old offspring following exposure to isolation in a novel environment or under resting conditions. Corticotropin-releasing factor titers in the hypothalamus were unaffected following isolation. Nonetheless, adrenocorticotropin hormone (ACTH) was found to be lower in the gestationally stressed offspring prior to or following the isolation period. Corticosterone was attenuated in gestationally stressed offspring following the postnatal stressor and there was also a tendency for the gestationally stressed females to have lower concentrations of aldosterone. Plasmatic testosterone levels were higher in the gestationally stressed males following the period of isolation. The present data suggest that the HPA axis of the offspring is differentially affected by the gestational stress procedure, that is, it is attenuated at the level of the pituitary and adrenal, but not at the level of the hypothalamus. These data have implications for behavioral differences observed in gestationally stressed animals.

Short- and Long-Term Consequences of Corticotropin-Releasing Factor in Early Development

Abstract: Corticotropin‐releasing factor (CRF) mediates various stress‐related responses in adult animals. Little is known about the effects of CRF during early development. Young mammals often vocalize when isolated in novel surroundings. Heightened levels of CRF inhibit vocalizing in isolated rat and guinea pig pups. Still lower levels of CRF may facilitate or permit vocalizing in rat pups. In guinea pigs, CRF appears to move pups from an initial active, to a subsequent passive, stage of behavioral responsiveness. CRF activity prior to birth can also affect the young. Exposing pregnant female rats to stressors during the last trimester of pregnancy alters the morphological and behavioral development of the offspring. Effects of gestational stress can be mimicked by injecting pregnant females with CRF during the last trimester. CRF appears to mediate both short‐ and long‐term responses to stressors during development in rodents.

The Distribution of Radiolabeled Corticotropin-Releasing Factor in Pregnant Rats: An Investigation of Placental Transfer to the Fetuses

Stress during gestation can have serious consequences on the development of the fetus. Many of these effects appear to be mediated by hormones of the hypothalamic‐pituitary‐adrenal (HPA) axis. Corticotropin‐releasing factor (CRF), released by the hypothalamus during times of stress serves to activate release of pituitary hormones and is also present in low levels in rat plasma. Moreover, the uterus contains significant quantities of CRF at implantation sites, probably from local sources. Therefore, the possibility exists that CRF may cross the placenta and activate the fetal HPA axis. However, the ability of CRF to cross the placenta has not been demonstrated. In the present study, pregnant rats were administered radiolabeled CRF intraperitoneally, and the distribution of the labeled product was determined in the fetuses and various maternal organs. High levels of activity were observed in the pregnant females uterus, adrenals, heart and the placentae, but only background levels of activity were detected in the maternal brain. Very low levels of activity were observed in the fetuses, indicating that the transfer of CRF across the placenta is greatly restricted. These findings suggest that maternal CRF has little or no direct effect on the developing fetus during gestational stress.

Some effects of methylphenidate on self-punitive running in rats

Self-punitive behavior was demonstrated in prepunishment speeds during extinction following shock-escape training in a straight runway under no-dose, low-dose (1-mg/kg), and high-dose (10-mg/kg) methylphenidate conditions. Increased dosage enhanced punished running (shock in the last half of the runway) and nonpunished running. Self-punitive behavior, defined as the difference between punishment conditions with faster running or increased persistence under punishment conditions, was not significantly affected by the drug variable. The results were interpreted as compatible with conditioned fear interpretations of self-punitive behavior but nonsupportive of cognitive interpretations.