Cardinal role of the environment in stress induced changes across life stages and generations.

The stress response in rodents and humans is exquisitely dependent on the environmental context. The interactive element of the environment is typically studied by creating laboratory models of stress-induced plasticity manifested in behavior or the underlying neuroendocrine mediators of the behavior. Here, we discuss three representative sets of studies where the role of the environment in mediating stress sensitivity or stress resilience is considered across varying windows of time. Collectively, these studies testify that environmental variation at an earlier time point modifies the relationship between stressor and stress response at a later stage. The metaplastic effects of the environment on the stress response remain possible across various endpoints, including behavior, neuroendocrine regulation, region-specific neural plasticity, and regulation of receptors. The timescale of such variation spans adulthood, across stages of life history and generational boundaries. Thus, environmental variables are powerful determinants of the observed diversity in stress response. The predominant role of the environment suggests that it is possible to promote stress resilience through purposeful modification of the environment.

Prefrontal-hippocampus plasticity reinstated by an enriched environment during stress.

Chronic stress causes dendritic atrophy of neurons within the hippocampus and medial prefrontal cortex. In this report, we show that chronic stress leads to reduced long-term potentiation in the pathway from the hippocampus to the medial prefrontal cortex of rats; and that such reduction is rescued by enriched housing environment. Connectivity between the hippocampus and medial prefrontal cortex is proposed to be an essential substrate that is often compromised in several psychiatric disorders. Our observations suggest that a short period of complexity in the housing environment has the potential to protect the functional integrity of this important connection.

Author Correction: Early­life short­term environmental enrichment counteracts the effects of stress on anxiety­like behavior, brain­derived neurotrophic factor and nuclear translocation of glucocorticoid receptors in the basolateral amygdala.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Female rats are resilient to the behavioral effects of maternal separation stress and exhibit stress-induced neurogenesis.

Early-life stress causes anxiogenesis and sensitivity of stress endocrine axis, facilitated by changes in the basolateral amygdala and hippocampal neurogenesis. In this report, we examined if male-like relationship between early-life stress and anxiety was recapitulated in female rats, along with related neurobiological substrates of the amygdala and the hippocampus. Maternal separation, a paradigm consistently utilized in male rats in most previously published scripts, did not cause similar behavioral consequences in females. Maternal separation caused an increase in adult hippocampal neurogenesis in females without causing substantial differences in dendritic arbors of the basolateral amygdala. Thus, female rats displayed remarkable resilience in the emotional consequences of early-life stress.

Early-life short-term environmental enrichment counteracts the effects of stress on anxiety-like behavior, brain-derived neurotrophic factor and nuclear translocation of glucocorticoid receptors in the basolateral amygdala.

Early life is a decisive stage for the development of physiological and psychological characteristics of an individual. Any stress or disruption of healthy development at this stage has serious long-lasting consequences for the remaining life. Unfortunately, early life stress is a common occurrence in humans and other animals. In this context, we investigated if the provision of environmental enrichment during the pre-weaning phase of rat pups and dams could alter the consequences of early-life maternal-separation stress. Pre-weaning enrichment rescued the effects of maternal separation on the excess secretion of adrenal stress hormones and anxiety-like behavior during adulthood. Enrichment also reduced the effect of stress on the spine density of basolateral amygdala neurons, a brain region critical for stress-induced facilitation of emotional behaviors. Pre-weaning enrichment, provided during early-life, blunted the effects of maternal separation stress on decreased intra-nuclear translocation of glucocorticoid receptors within the amygdala neurons when tested later in adulthood. Early-life, pre-weaning environmental enrichment also increased the amount of brain-derived neurotrophic factor within adult basolateral amygdala. Our observations showed that environmental manipulation during early formative years could be utilized to build lifelong resilience to stress. Complex naturalistic housing and sensory enrichment is, thus, an useful buffer against an impoverished and stressful childhood.

Environment and early life: Decisive factors for stress-resilience and vulnerability.

Early life is a critical and sensitive period whereby environmental imprints on later life are generated. These environmental influences from early life have long-lasting consequences on for mental health. Both human and animal work suggests that maternal presence constitutes an important part of the early experience. Maternal separation causes a sustained increase in stress responsiveness later in life, along with facilitated anxiety-like behaviors. On the contrary, providing a complex and enriching sensory environment during or after stress, on the other hand, creates resilience to stress. In this chapter, we summarize these environmental influences on the maternal interactions and subsequent stress susceptibility or resilience of the offspring.

Neuronal Plasticity in the Amygdala Following Predator Stress Exposure.

Predation causes robust long-term stress-related effects on prey individuals even if they do not get consumed by the predator. Here I review the role of basolateral amygdala (BLA) neurons in the mediation of non-consumptive effects of predation. This brain region is critical for the generation and maintenance of fear response across many phylogenetic groups. The exposure to cues of predator presence activates neurons within the BLA. Hormones secreted during stressful episodes cause long-lasting structural changes in BLA neurons, causing facilitation of endocrine response during subsequent exposure to stressful episodes like later predator exposure. Some studies also suggest that BLA is involved in creating anticipatory defensive behavior in response to the expectation of change in the environment.

Enriched Environment Facilitates Anxiolytic Efficacy Driven by Deep-Brain Stimulation of Medial Prefrontal Cortex.

Deep brain stimulation (DBS) is a widely used treatment for neurodegenerative disorders like Parkinson's disease. Recently, several studies have used preclinical animal models to suggest that DBS has a potential to improve emotional symptoms in mental disorders such as treatment-resistant depression and post-traumatic stress disorder. An important difference between neurodegenerative and emotional disorders is the crucial role of environment in the ontogeny of the latter. Thus, it is important to understand the effects of DBS in the context of environmental variation. In this study, we show that DBS of ventromedial prefrontal cortex reduces anxiety in rats when it is coupled with simultaneous exposure to an enriched environment (EE). In contrast, effects of DBS on anxiety-like behaviors remained equivocal when animals were housed in standard laboratory conditions. These results suggest that the ability of DBS to treat anxiety and related phenotypes can be significantly enhanced by EE opportunities.

Complex housing causes a robust increase in dendritic complexity and spine density of medial prefrontal cortical neurons.

Prelimbic cortex and infralimbic cortex, parts of the ventromedial prefrontal cortex, are critical brain regions for generating a flexible behavioral response to changing environmental contingencies. This includes the role of these brain structures in the extinction of learned fear, decision making and retrieval of remote memories. Dendritic structure of medial prefrontal cortex neurons retains significant structural plasticity in adulthood. This has been mainly demonstrated as dendritic atrophy and loss of dendritic spines due to chronic stress. It remains unknown if housing condition of the animals itself can cause opposing changes in the dendritic organization. In that backdrop, here we report that short-term increase in complexity of the housing causes a robust increase in complexity of dendritic architecture of prelimbic and infralimbic neurons. This is reflected in the dendritic expansion of prelimbic neurons and increase in spine density of prelimbic and infralimbic neurons. These results suggest that non-invasive changes in the housing environment can be harnessed to study brain reserves for the flexible and species-typical behaviors.

Housing environment influences stress-related hippocampal substrates and depression-like behavior.

Rats are widely used animal models for biological psychiatry and neuroscience. Laboratory rats are typically housed in impoverished sensory environments. The lack of species-typical sensory environment might radically change the response of individual animals to stressful and/or threatening episodes. In this report, we demonstrate that behavioral and neural sequelae of chronic stress were modified by sensory environment of adult male rats. This includes effects of stress on the density of spines on CA3 hippocampal neurons, hippocampal neurogenesis and abundance of glucocorticoid or mineralocorticoid receptors. Enrichment also reduced depression-like behavior in a forced swim task. Stress and sensory enrichment evoked opposing effects on all the above endpoints. The sensory enrichment used in this report is of a relatively short duration provided during adulthood. This period excludes critical windows of greater plasticity during pre- and peripubertal stages. Our results suggest that standard housing practices for laboratory rats remain austere concerning sensory requirements of this species. Thus, even a moderate sensory enrichment is capable of reducing high stress-sensitivity and depressive-like behavior in standard laboratory rats.

Dendritic Architecture of Principal Basolateral Amygdala Neurons Changes Congruently with Endocrine Response to Stress.

Animals cope with changing environments through changes in behavior. Such plasticity is, however, marked by substantial inter-individual variability. Neuroendocrine reactivity to challenging environments can be an important predictor of resilience. Both basolateral amygdala (BLA) neurons and adrenal glucocorticoid signaling are integral parts of the stress neuroendocrine response. In this report, we test if individual variation in hormonal response to stress is associated with individual variation in the dendritic complexity of BLA neurons. We report a positive correlation between inter-individual variability in glucocorticoid response and neuronal plasticity in the BLA subsequent to a stressor. This suggests that stressful experiences in the past act as significant sculptors of BLA neuronal plasticity and congruent neuroendocrine response.

Effects of stress or infection on rat behavior show robust reversals due to environmental disturbance.

Background: The behavior of animals is intricately linked to the environment; a relationship that is often studied in laboratory conditions by using environmental perturbations to study biological mechanisms underlying the behavioral change.  Methods: This study pertains to two such well-studied and well-replicated perturbations, i.e., stress-induced anxiogenesis and Toxoplasmagondii -induced loss of innate fear. Here, we demonstrate that behavioral outcomes of these experimental manipulations are contingent upon the ambient quality of the wider environment where animal facilities are situated. Results: During late 2014 and early 2015, a building construction project started adjacent to our animal facility. During this phase, we observed that maternal separation stress caused anxiolysis, rather than historically observed anxiogenesis, in laboratory rats. We also found that Toxoplasma gondii infection caused an increase, rather than historically observed decrease, in innate aversion to predator odors in rats. Conclusion: These observations suggest that effects of stress and Toxoplasma gondii are dependent on variables in the environment that often go unreported in the published literature.

Short-term environmental enrichment is sufficient to counter stress-induced anxiety and associated structural and molecular plasticity in basolateral amygdala.

Moderate levels of anxiety enable individual animals to cope with stressors through avoidance, and could be an adaptive trait. However, repeated stress exacerbates anxiety to pathologically high levels. Dendritic remodeling in the basolateral amygdala is proposed to mediate potentiation of anxiety after stress. Similarly, modulation of brain-derived neurotrophic factor is thought to be important for the behavioral effects of stress. In the present study, we investigate if relatively short periods of environmental enrichment in adulthood can confer resilience against stress-induced anxiety and concomitant changes in neuronal arborisation and brain derived neurotrophic factor within basolateral amygdala. Two weeks of environmental enrichment countermanded the propensity of increased anxiety following chronic immobilization stress. Environmental enrichment concurrently reduced dendritic branching and spine density of projection neurons of the basolateral amygdala. Moreover, stress increased abundance of BDNF mRNA in the basolateral amygdala in agreement with the dendritic hypertrophy post-stress and role of BDNF in promoting dendritic arborisation. In contrast, environmental enrichment prevented stress-induced rise in the BDNF mRNA abundance. Gain in body weights and adrenal weights remained unaffected by exposure to environmental enrichment. These observations suggest that a short period of environmental enrichment can provide resilience against maladaptive effects of stress on hormonal, neuronal and molecular mediators of anxiogenesis.

Seeding Stress Resilience through Inoculation.

Stress is a generalized set of physiological and psychological responses observed when an organism is placed under challenging circumstances. The stress response allows organisms to reattain the equilibrium in face of perturbations. Unfortunately, chronic and/or traumatic exposure to stress frequently overwhelms coping ability of an individual. This is manifested as symptoms affecting emotions and cognition in stress-related mental disorders. Thus environmental interventions that promote resilience in face of stress have much clinical relevance. Focus of the bulk of relevant neurobiological research at present remains on negative aspects of health and psychological outcomes of stress exposure. Yet exposure to the stress itself can promote resilience to subsequent stressful episodes later in the life. This is especially true if the prior stress occurs early in life, is mild in its magnitude, and is controllable by the individual. This articulation has been referred to as "stress inoculation," reminiscent of resilience to the pathology generated through vaccination by attenuated pathogen itself. Using experimental evidence from animal models, this review explores relationship between nature of the "inoculum" stress and subsequent psychological resilience.

Toxoplasma gondii infection induces dendritic retraction in basolateral amygdala accompanied by reduced corticosterone secretion.

Pathological anxiety is thought to reflect a maladaptive state characterized by exaggerated fear. Naturally occurring perturbations that reduce fear can be crucial in the search for new treatments. The protozoan parasite Toxoplasma gondii invades rat brain and removes the fear that rats have of cat odors, a change believed to be parasitic manipulation of host behavior aimed at increasing parasite transmission. It is likely that mechanisms employed by T. gondii can be used as a heuristic tool to understand possible means of fear reduction in clinical settings. Male Long-Evans rats were infected with T. gondii and compared with sham-infected animals 8 weeks after infection. The amount of circulating plasma corticosterone and dendritic arborization of basolateral amygdala principal neurons were quantified. Previous studies have shown that corticosterone, acting within the basolateral amygdala, enhances the fear response to environmental stimuli. Here we show that T. gondii infection causes a dendritic retraction in basolateral amygdala neurons. Such dendritic retraction is accompanied by lower amounts of circulating corticosterone, both at baseline and when induced by an aversive cat odor. The concerted effects of parasitism on two pivotal physiological nodes of the fear response provide an animal model relevant to interactions between stress hormones and amygdalar plasticity.

Short-term enrichment makes male rats more attractive, more defensive and alters hypothalamic neurons.

Innate behaviors are shaped by contingencies built during evolutionary history. On the other hand, environmental stimuli play a significant role in shaping behavior. In particular, a short period of environmental enrichment can enhance cognitive behavior, modify effects of stress on learned behaviors and induce brain plasticity. It is unclear if modulation by environment can extend to innate behaviors which are preserved by intense selection pressure. In the present report we investigate this issue by studying effects of relatively short (14-days) environmental enrichment on two prominent innate behaviors in rats, avoidance of predator odors and ability of males to attract mates. We show that enrichment has strong effects on both the innate behaviors: a) enriched males were more avoidant of a predator odor than non-enriched controls, and had a greater rise in corticosterone levels in response to the odor; and b) had higher testosterone levels and were more attractive to females. Additionally, we demonstrate decrease in dendritic length of neurons of ventrolateral nucleus of hypothalamus, important for reproductive mate-choice and increase in the same in dorsomedial nucleus, important for defensive behavior. Thus, behavioral and hormonal observations provide evidence that a short period of environmental manipulation can alter innate behaviors, providing a good example of gene-environment interaction.

Stress coping stimulates hippocampal neurogenesis in adult monkeys.

Coping with intermittent social stress is an essential aspect of living in complex social environments. Coping tends to counteract the deleterious effects of stress and is thought to induce neuroadaptations in corticolimbic brain systems. Here we test this hypothesis in adult squirrel monkey males exposed to intermittent social separations and new pair formations. These manipulations simulate conditions that typically occur in male social associations because of competition for limited access to residency in mixed-sex groups. As evidence of coping, we previously confirmed that cortisol levels initially increase and then are restored to prestress levels within several days of each separation and new pair formation. Follow-up studies with exogenous cortisol further established that feedback regulation of the hypothalamic-pituitary-adrenal axis is not impaired. Now we report that exposure to intermittent social separations and new pair formations increased hippocampal neurogenesis in squirrel monkey males. Hippocampal neurogenesis in rodents contributes to spatial learning performance, and in monkeys we found that spatial learning was enhanced in conditions that increased hippocampal neurogenesis. Corresponding changes were discerned in the expression of genes involved in survival and integration of adult-born granule cells into hippocampal neural circuits. These findings support recent indications that stress coping stimulates hippocampal neurogenesis in adult rodents. Psychotherapies designed to promote stress coping potentially have similar effects in humans with major depression.

Gene therapy in rodent amygdala against fear disorders.

IMPORTANCE OF THE FIELD: Successful treatment of fear and anxiety disorders is presently a difficult task. A major limitation is the fact that underlying physiological mechanisms of fear and anxiety are only now beginning to be understood. As we obtain more information about mechanisms and brain circuits involved, treatment of these conditions will become increasingly realistic. AREAS COVERED IN THIS REVIEW: Gene therapy is a promising treatment strategy that has several advantages over the more widely used pharmacological approaches. In this review we discuss the potential and limitation of gene therapy in the amygdala. This review concerns itself with papers published within the last 20 years in the field of gene therapy for fear and anxiety, and has been primarily conducted in rodent models. WHAT THE READER WILL GAIN: We present the current state of research into gene therapy for anxiety disorders, using a few case studies. The review will delineate challenges, limitations and opportunities for successful gene therapy. TAKE HOME MESSAGE: There are sizeable gains in knowledge about functioning of the fear system. This will inform future improvements in gene therapy approaches. An ideal gene therapy strategy will involve improvements in both delivery vectors and in design of therapeutic cargo.

Expression of chimeric estrogen-glucocorticoid-receptor in the amygdala reduces anxiety.

The basolateral amygdala is critical for generating anxiety, and exposure to glucocorticoids induces anxiety. The demonstrated ability of glucocorticoids to cause dendritic expansion and increase excitability in the amygdala could help mediate the behavioral effects of glucocorticoids, and thus may be important therapeutic target for anxiety. In contrast, estrogen has the opposite effects of glucocorticoids in many domains. In this study, we employed a gene therapeutic approach to reduce anxiety and dendritic arborization of amygdaloid neurons of adult male Wistar rats. We used a herpes simplex viral vector to express a chimeric steroid receptor ("ERGR") which binds glucocorticoids yet transduces their actions into estrogenic effects. When expressed in the basolateral amygdala (BLA), ERGR reduced anxiety, as tested on elevated plus-maze and open field, without affecting conditioned fear, another amygdala-dependent behavior. Moreover, ERGR also blocked glucocorticoid-induced dendritic expansion of BLA neurons. Thus ERGR expression in the BLA provides a potential therapeutic against anxiety disorders.

Resilience against predator stress and dendritic morphology of amygdala neurons.

Individual differences in coping response lie at the core of vulnerability to conditions like post-traumatic stress disorder (PTSD). Like humans, not all animals exposed to severe stress show lasting change in affect. Predator stress is a traumatic experience inducing long-lasting fear, but not in all rodents. Thus, individual variation may be a cross species factor driving responsiveness to stressful events. The present study investigated neurobiological bases of variation in coping with severe stress. The amygdala was studied because it modulates fear and its function is affected by stress. Moreover, stress-induced plasticity of the amygdala has been related to induction of anxiety, a comorbid symptom of psychiatric conditions like PTSD. We exposed rodents to predator stress and grouped them according to their adaptability based on a standard anxiety test (the elevated plus maze). Subsequently we investigated if well-adapted (less anxious) and mal-adapted (extremely anxious) stressed animals differed in the structure of dendritic trees of their output neurons of the right basolateral amygdala (BLA). Two weeks after exposure to stress, well-adapted animals showed low anxiety levels comparable to unstressed controls, whereas mal-adapted animals were highly anxious. In these same animals, Golgi analysis revealed that BLA neurons of well-adapted rats exhibited more densely packed and shorter dendrites than neurons of mal-adapted or unstressed control animals, which did not differ. These data suggest that dendritic hypotrophy in the BLA may be a resilience marker against lasting anxiogenic effects of predator stress.