Christoph P. Wiedenmayer

Adaptations or pathologies? Long-term changes in brain and behavior after a single exposure to severe threat

The experience of a single threatening situation may alter the behavior of an animal in a long-lasting way. Long-lasting changes in behavior have been induced in laboratory animals to model and investigate the development and neural substrate of human psychopathologies. Under natural conditions, however, changes in behavior after an aversive experience may be adaptive because behavioral modifications allow animals to adjust to a threat for extended periods of time. In the laboratory setting, properties of the aversive situation and the potential of the animal to respond to the threat may be altered and lead to extensive, prolonged changes, indicating a failure in behavioral regulation. Such long-term changes seem to be mediated by neuronal alterations in components of the fear pathway. To understand psychopathologies, determinants of exaggerated responsivity and the underlying molecular and neural processes have to be analyzed in a comparative way under conditions that produce normal and abnormal fear and anxiety.

The Role of the Medial Prefrontal Cortex in Innate Fear Regulation in Infants, Juveniles, and Adolescents

In adult animals, the medial prefrontal cortex (mPFC) plays a significant role in regulating emotions and projects to the amygdala and periaqueductal gray (PAG) to modulate emotional responses. However, little is known about the development of this neural circuit and its relevance to unlearned fear in pre-adulthood. To address these issues, we examined the mPFC of 14-d-old (infants), 26-d-old (juveniles), and 38- to 42-d-old (adolescents) rats to represent different developmental and social milestones. The expression patterns of the neuronal marker FOS were used to assess neurological activity. Muscimol, a GABA agonist, was used to inactivate the prelimbic and infralimbic mPFC subdivisions (400 ng in 200 nl). Animals were exposed to either a threatening or nonthreatening stimulus that was ecologically relevant and age specific. Freezing was measured as an indicator of innate fear behavior. The data indicated that the mPFC is neither active nor responsive to innate fear in infant rats. In juveniles, the prelimbic mPFC became responsive in processing aversive sensory stimulation but did not regulate freezing behavior. Finally, during adolescence, inactivation of the prelimbic mPFC significantly attenuated freezing and decreased FOS expression in the ventral PAG. Surprisingly, across all ages, there were no significant differences in FOS levels in the medial and basolateral/lateral amygdala when either mPFC subdivision was inactivated. Together, unlearned fear has a unique developmental course with different brain areas involved in unlearned fear in the immature animal than the adult. In particular, the mPFC neural circuitry is different in young animals and progressively develops more capacities as the animal matures.

Developmental changes in c-fos expression to an age-specific social stressor in infant rats.

Young rats become immobile when exposed to a potentially infanticidal adult male rat. Male-induced immobility declines during the preweaning period, paralleling the decrease in infanticidal threat. To investigate the neural substrates underlying the developmental change in immobility, male-induced expression of the immediate-early gene c-fos was assessed on postnatal days 7, 14 and 21. A huddle of three young rats was exposed to an adult male behind a screen. As control, three littermates were put in the testing chamber but not exposed to the male. On day 7, male exposed and control pups were immobile most of the time and c-fos expression did not differ between conditions. On day 14, rats in the presence of the male stopped ongoing behaviors and became immobile. They had significantly higher c-fos expression in the paraventricular nucleus of the hypothalamus, the amygdala, the periaqueductal gray, and the locus ceruleus. On day 21, the male-exposed rats that were immobile had elevated c-fos expression in a similar pattern as on day 14, however, different nuclei of the amygdala were activated. In contrast, male-exposed 21-day-old rats that showed control levels of immobility did not have elevated c-fos expression in these areas. These results demonstrate that male exposure induced c-fos expression in brain areas of young rats in an age-specific pattern. Some of the activated brain areas seem to have contributed to immobility. Differential activation of neuronal populations may underlie developmental changes in defensive immobility during early ontogeny.

Plasticity of defensive behavior and fear in early development

Animals have the ability to respond to threatening situations with sets of defensive behaviors. This review demonstrates that defensive behaviors change during early life in mammals. First, unlearned responses are reorganized during early ontogeny and expressed in an age-specific way. Second, the expression of defensive responses is influenced by early experience prior to the first encounter with a threat. Third, once animals have been exposed to a threatening stimulus they subsequently modify their behavior. The neural bases of defensive behavior and the processes that alter them during development are discussed. Maturation of components and connections of the fear circuit seem to contribute to changes in unlearned fear responses. Early experience and learning modify these developmental processes and shape the expression of defensive behavior. Continuous reorganization of the neural substrate and defensive behavior during ontogeny seems to allow the animal to adjust to the conditions it encounters at a given age in a given environment. It is proposed that the developmental changes in defensive behavior can be conceptualized as phenotypic plasticity.

The effect of periaqueductal gray lesions on responses to age-specific threats in infant rats.

During early ontogeny infant rats show specific responses to a variety of age-dependent threatening situations. When isolated from nest and dam, they emit ultrasonic vocalizations and show decreased reactivity to noxious stimulation, or analgesia. When exposed to an unfamiliar adult male, they become immobile and analgesic. The midbrain periaqueductal gray (PAG) is an important area within the circuitry that controls responses to threatening stimuli in the adult. Little is known about the functions of the PAG in early life. It was hypothesized that the PAG mediates the responses to the age-specific threats social isolation and male exposure in the infant rat. Rat pups were lesioned electrolytically either in the lateral or the ventrolateral PAG on postnatal day 7, tested in social isolation on day 10, and exposed to a male on day 14. On day 10 during isolation, ultrasonic vocalizations and isolation-induced analgesia were decreased in both lesion groups. On day 14, male-induced immobility and analgesia were decreased in ventrally lesioned animals. In conclusion, the PAG seems to play a developmentally continuous role in age-specific responses to threat such as ultrasonic vocalization, analgesia, and immobility.

μ opioid receptors in the ventrolateral periaqueductal gray mediate stress-induced analgesia but not immobility in rat pups

Rat pups become immobile and analgesic when exposed to an adult male rat. The aim of this study was to determine whether these reactions are under the control of endogenous opioids and to determine the role of the midbrain periaqueductal gray (PAG), which mediates stress-induced immobility and analgesia in adult animals. In Experiment 1, 14-day-old rats were injected systemically with the general opioid receptor antagonist naltrexone (1 mg/kg), which blocked male-induced analgesia to thermal stimulation but did not affect immobility. In Experiment 2, the selective mu opioid receptor antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP; 50 or 100 ng/200 nl) was microinjected into the ventrolateral and lateral PAG. CTOP suppressed male-induced analgesia when injected into the ventrolateral PAG. Male-induced immobility was not affected by CTOP. Male proximity therefore seems to induce analgesia in rat pups by releasing endogenous opioids that bind to mu opioid receptors in the ventrolateral PAG.

Cortisol Levels and Hippocampus Volumes in Healthy Preadolescent Children

BACKGROUND Research in animal models has demonstrated that elevated levels of glucocorticoids can inflict damage within the hippocampus. In adult humans, elevated cortisol levels have been associated with reduced hippocampal volumes; however, normative data in children are not available. The objective of this study was to examine possible associations of serum cortisol levels with hippocampal volumes and morphology in healthy children. METHODS Morning serum cortisol levels and hippocampus magnetic resonance imaging were measured in 17 healthy children (8 girls, 9 boys) between 7 and 12 years of age. RESULTS Cortisol levels were not associated with total hippocampal volumes; however, with an analysis of surface morphology, significant associations were found for regionally specific portions of the hippocampus. Positive associations were detected for the anterior segment of the hippocampus and inverse associations along the lateral aspects of the hippocampus. CONCLUSIONS Associations of cortisol levels with regionally specific variations in hippocampal morphology were detected during early development in healthy preadolescent children.

Mother Lowers Glucocorticoid Levels of Preweaning Rats After Acute Threat

Abstract: Exposure to a deadly threat, an adult male rat, induced the release of corticosterone in 14‐day‐old rat pups. The endocrine stress response was decreased when the pups were reunited with their mother immediately after exposure. These findings demonstrate that social variables can reduce the consequences of an aversive experience.

The Role of the Amygdala and Olfaction in Unconditioned Fear in Developing Rats

Early in ontogeny, young rats must be able to detect dangerous stimuli and to exhibit appropriate defensive behaviors. Different nuclei of the amygdala mediate unconditioned and conditioned fear responses to threat in adult rats. The aim of this study was to determine the role of the amygdala in unlearned fear behavior in young rats. When exposed to an unfamiliar adult male rat, preweaning rat pups freeze, with peak levels on postnatal day 14 and declining levels on day 18. Pups were made anosmic to block olfactory input to the amygdala, and amygdala activation was assessed by quantifying the neuronal marker c-fos. Anosmic pups did not freeze in the presence of the male rat and had decreased c-fos expression in the medial amygdala on day 14 and in the medial and lateral amygdala on day 18. However, the decrease in freezing between days 14 and 18 was not associated with a decrease in c-fos expression in the medial amygdala. The medial and lateral amygdala were then inactivated by local muscimol infusion on day 14. Muscimol infusion into the medial amygdala decreased freezing to the male rat but not to a loud noise, whereas infusion into the lateral amygdala blocked freezing to a loud noise but not to the male. These findings indicate that different nuclei of the amygdala process sensory information of different modalities, mediate unconditioned freezing, and may be involved in developmental changes in the fear response in young rats.

Age-specific threats induce CRF expression in the paraventricular nucleus of the hypothalamus and hippocampus of young rats.

Young animals respond to threatening stimuli in an age-specific way. Their endocrine and behavioral responses reflect the potential threat of the situation at a given age. The aim of the present study was to determine whether corticotropin-releasing factor (CRF) is involved in the endocrine and behavioral responses to threat and their developmental changes in young rats. Preweaning 14-day-old and postweaning 26-day-old rats were exposed to two age-specific threats, cat odor and an adult male rat. The acute behavioral response was determined during exposure. After exposure, the time courses of the corticosterone response and of CRF expression in the paraventricular nucleus of the hypothalamus (PVN) and in extrahypothalamic areas were assessed. Preweaning rats became immobile when exposed to cat odor or the male rat, whereas postweaning rats became immobile to cat odor only. Male exposure increased serum corticosterone levels in 14-day-old rats, but cat odor failed to increase levels at either age. Exposure induced elevation of CRF mRNA levels in the PVN that paralleled changes in corticosterone levels. CRF may thus play a role in endocrine regulation and its developmental changes during early life. Neither cat odor nor the adult male altered CRF mRNA levels in the bed nucleus of the stria terminalis (BNST) or the amygdala, but both stimuli increased levels in the hippocampus. Hippocampal CRF mRNA expression levels did not parallel cat odor or male-induced immobility, indicating that CRF is not involved in this response in young rats but may be involved in aspects of learning and memory.