Captivity alters neuroendocrine regulators of stress and reproduction in the hypothalamus in response to acute stress.

Wild animals are brought into captivity for many reasons. However, unlike laboratory-bred animals, wild caught animals often respond to the dramatic shift in their environment with physiological changes in the stress and reproductive pathways. Using wild-caught male and female house sparrows (Passer domesticus) we examined how time in captivity affects the expression of reproductive and stress-associated genes in the brain, specifically, the hypothalamus. We quantified relative mRNA expression of a neurohormone involved in the stress response (corticotropin releasing hormone [CRH]), a hypothalamic inhibitor of reproduction (gonadotropin inhibitory hormone [GnIH]), and the glucocorticoid receptor (GR), which is important in terminating the stress response. To understand potential shifts at the cellular level, we also examined the presence of hypothalamic GnIH (GnIH-ir) using immunohistochemistry. We hypothesized that expression of these genes and the abundance of cells immunoreactive for GnIH would change in response to time in captivity as compared to free-living individuals. We found that GR mRNA expression and GnIH-ir cell abundance increased after 24 and 45 days in captivity, as compared to wild-caught birds. At 66 days in captivity, GR expression and GnIH cell abundance did not differ from wild-caught birds, suggesting birds had acclimated to captivity. Evaluation of CRH and GnIH mRNA expression yielded similar trends, though they were not statistically significant. In addition, although neuroendocrine factors appeared to acclimate to captivity, a previous study indicated that corticosterone release and immune responses of these same birds did not acclimate to captivity, suggesting that neuroendocrine endpoints may adapt more rapidly to captivity than downstream physiological measures. These data expand our understanding of the physiological shifts occurring when wild animals are brought into captivity.

Moving Forward From COVID-19: Bridging Knowledge Gaps in Maternal Health With a New Conceptual Model.

As the world faces the health crisis of a global pandemic-with healthcare protocols in overhaul, and patients and care teams experiencing unprecedented levels of stress and unpredictability-we predict that current knowledge gaps in maternal health will inevitably have a lasting impact on the health of women giving birth now and in the near future. Since we are decades away from closing the knowledge gaps we need filled today, we recommend shifting thinking toward a comprehensive conceptual model that merges knowledge of stress physiology, neurobiology, and pregnancy physiology. The model we present here, the Maternal Reactive Scope Model, is an expansion of the Reactive Scope Model built upon the concept of Homeostasis and Allostasis. The model provides a framework to consider pathways and interactions across physiological systems to attribute a physiological basis for considering stress exposure and bridge research gaps on mechanisms to measure or target for treatment. Our intention is to provide an adaptable, heuristic framework for discussion of research considerations and new healthcare models that aim to provide the best care for new mothers during and after the COVID-19 pandemic.

The HPA Axis During the Perinatal Period: Implications for Perinatal Depression.

The transition to motherhood is characterized by some of the most pronounced endocrine changes a woman will experience in her lifetime. Unfortunately, matrescence is also a time in a woman's life when she is most susceptible to mental illness such as perinatal depression. A growing body of research has aimed to determine how key endocrine systems, such as the hypothalamic-pituitary-adrenal (HPA) axis, are involved in the dysregulation of perinatal mental health. However, very little research has consistently linked perinatal changes in the HPA axis with maternal mental illness. Therefore, the aims of this mini review are to: (i) clearly summarize the normative changes in the HPA axis that occur during pregnancy and the postpartum period; (ii) summarize what we know about the HPA axis in perinatal depression, and (iii) propose key areas for future research. Understanding physiological biomarkers that can predict which women are at risk for perinatal mood disorders will lead to better tools for treating, and ultimately preventing, these debilitating disorders, improving the health of mother, child, and family.

Neural Versus Gonadal GnIH: Are they Independent Systems? A Mini-Review.

Based on research in protochordates and basal vertebrates, we know that communication across the first endocrine axes likely relied on diffusion. Because diffusion is relatively slow, rapid responses to some cues, including stress-related cues, may have required further local control of axis outputs (e.g., steroid hormone production by the gonads). Despite the evolution of much more efficient circulatory systems and complex nervous systems in vertebrates, production of many "neuro"transmitters has been identified outside of the hypothalamus across the vertebrate phylogeny and these neurotransmitters are known to locally regulate endocrine function. Our understanding of tissue-specific neuropeptide expression and their role coordinating physiological/behavioral responses of the whole organism remains limited, in part, due to nomenclature and historic dogma that ignores local regulation of axis output. Here, we review regulation of gonadotropin-inhibitory hormone (GnIH) across the reproductive axis in birds and mammals to bring further attention to context-dependent disparities and similarities in neuropeptide production by the brain and gonads. We find that GnIH responsiveness to cues of stress appears conserved across species, but that the response of specific tissues and the direction of GnIH regulation varies. The implications of differential regulation across tissues remain unclear in most studies, but further work that manipulates and contrasts function in different tissues has the potential to inform us about both organism-specific function and endocrine axis evolution.

Endocannabinoid Signaling in the Stress Response of Male and Female Songbirds.

Endocannabinoid (eCB) signaling plays an important role in the stress response pathways of the mammalian brain, yet its role in the avian stress response has not been described. Understanding eCB signaling in avian species (such as the European starling, Sturnus vulgaris) allows a model system that exhibits natural attenuation of hypothalamic-pituitary-adrenal (HPA) responsiveness to stressors. Specifically, seasonally breeding birds exhibit the highest HPA activity during the breeding season and subsequently exhibit a robust HPA down-regulation during molt. Because eCB signaling in mammals has an overall inhibitory effect on HPA activity, we expected shifts in eCB signaling to regulate the seasonal HPA down-regulation during molt. However, our data did not support a role for eCB signaling in the molt-related suppression of HPA activity. For example, injection of the cannabinoid receptor (CB1) antagonist, AM251, did not potentiate molt-suppressed HPA activity. Instead, our data suggest eCB regulation of HPA plasticity as birds transition from breeding to molt. In support of this hypothesis, birds in the late breeding season demonstrated a more dynamic response at the level of avian amygdala eCB content in response to acute stress. The response and directionality of this effect match that seen in mammals. Overall, our data suggest that eCB signaling may allow for a dynamic range in HPA responsiveness (eg, breeding), but the signaling pathway's role may be limited when the HPA response is restrained (eg, molt). This first characterization of eCB signaling in the avian stress response also emphasizes that although the system functions similarly to other species, its exact role may be species specific.

Stress, captivity, and reproduction in a wild bird species.

In seasonal species, glucocorticoid concentrations are often highest during the breeding season. However, the role of increased hypothalamic-pituitary-adrenal (HPA) activity in the regulation of reproduction remains poorly understood. Our study is the first, to our knowledge, to document reproductive consequences of a non-pharmacological hindrance to seasonal HPA fluctuations. Using wild-caught male and female European starlings (Sturnus vulgaris) housed in an outdoor, semi-natural environment, we divided birds into two mixed-sex groups. One group remained in the outdoor aviary, where starlings breed at the appropriate time of year. The other group was transferred into an indoor flight aviary, where we predicted reproductive suppression to occur. We measured changes in corticosterone (CORT) at baseline and stress-induced concentrations prior to group separation and at the experiment's conclusion. After ten days, the birds showed remarkable differences in breeding behavior and HPA activity. Outdoor birds exhibited increases in baseline and stress-induced CORT and progressed into active breeding (pairing, nest building, egg laying, etc.). In contrast, indoor birds displayed no change in baseline or stress-induced CORT and few signs of active breeding. We found significant sex and treatment effects on expression of HPA and hypothalamic-pituitary-gonadal (HPG) axis elements, suggesting sex-specific regulatory mechanisms. Our data suggest a novel, facilitating role for the HPA axis in the transition between early breeding and active breeding in a wild, seasonal avian species. In addition, understanding how changes in housing condition affect seasonal HPA fluctuations may help alleviate barriers to breeding wild animals in captivity.

A consensus endocrine profile for chronically stressed wild animals does not exist.

Given the connection between chronic stress and health, there has been a growing emphasis on identifying chronically stressed wild animals, especially in relation to anthropogenic disturbances. There is considerable confusion, however, in how to identify chronically stressed wild animals, but the most common assumption is that measures of glucocorticoid (GC) function will increase. In an attempt to determine an "endocrine profile" of a chronically stressed wild animal, this review collected papers from the literature that measured baseline GC, stress-induced GC, measures of integrated GC, negative feedback, hypothalamic-pituitary-adrenal axis sensitivity, and/or body weight in chronically stressed animals. The collected studies encompassed laboratory and field studies, numerous diverse species, and multiple techniques for inducing chronic stress. Each paper was ranked according to its relevance to wild animals and scored as to whether the measured response increased, decreased, or stayed the same after exposure to chronic stress. The analyses uncovered so much variation between studies that the literature does not support a generalized endocrine profile in how wild animals respond to chronic stress. The common predictions appear to be based almost entirely on theoretical models rather than empirical data. The three most important variables affecting GC responses were the stressors used to induce chronic stress, the potential for those stressors to induce habituation, and the taxon of the focal species. The best approach for identifying a chronically stressed population appears to be documentation of changes at multiple levels of GC regulation, but the direction of the change (increase or decrease) may be relatively unimportant compared to the fact that the response changes at all. The conclusion is that a consistent, predictable, endocrine response to chronic stress, regardless of the protocol used to induce chronic stress and the species under study, does not exist.

Dynamic changes in brain aromatase activity following sexual interactions in males: where, when and why?

It is increasingly recognized that estrogens produce rapid and transient effects at many neural sites ultimately impacting physiological and behavioral endpoints. The ability of estrogens to acutely regulate cellular processes implies that their concentration should also be rapidly fine-tuned. Accordingly, rapid changes in the catalytic activity of aromatase, the limiting enzyme for estrogen synthesis, have been identified that could serve as a regulatory mechanism of local estrogen concentrations. However, the precise anatomical localization, time-course, triggering stimuli and functional significance of these enzymatic changes in vivo are not well understood. To address these issues as to where, when and why aromatase activity (AA) rapidly changes after sexual interactions, AA was assayed in six populations of aromatase-expressing cells microdissected from the brain of male quail that experienced varying durations of visual exposure to or copulation with a female. Sexual interactions resulted in a rapid AA inhibition. This inhibition occurred in specific brain regions (including the medial preoptic nucleus), in a context-dependent fashion and time-scale suggestive of post-translational modifications of the enzyme. Interestingly, the enzymatic fluctuations occurring in the preoptic area followed rather than preceded copulation and were tied specifically to the female's presence. This pattern of enzymatic changes suggests that rapid estrogen effects are important during the motivational phase of the behavior to trigger physiological events essential to activate mate search and copulation.

Acute stress differentially affects aromatase activity in specific brain nuclei of adult male and female quail.

The rapid and temporary suppression of reproductive behavior is often assumed to be an important feature of the adaptive acute stress response. However, how this suppression operates at the mechanistic level is poorly understood. The enzyme aromatase converts testosterone to estradiol in the brain to activate reproductive behavior in male Japanese quail (Coturnix japonica). The discovery of rapid and reversible modification of aromatase activity (AA) provides a potential mechanism for fast, stress-induced changes in behavior. We investigated the effects of acute stress on AA in both sexes by measuring enzyme activity in all aromatase-expressing brain nuclei before, during, and after 30 min of acute restraint stress. We show here that acute stress rapidly alters AA in the male and female brain and that these changes are specific to the brain nuclei and sex of the individual. Specifically, acute stress rapidly (5 min) increased AA in the male medial preoptic nucleus, a region controlling male reproductive behavior; in females, a similar increase was also observed, but it appeared delayed (15 min) and had smaller amplitude. In the ventromedial and tuberal hypothalamus, regions associated with female reproductive behavior, stress induced a quick and sustained decrease in AA in females, but in males, only a slight increase (ventromedial) or no change (tuberal) in AA was observed. Effects of acute stress on brain estrogen production, therefore, represent one potential way through which stress affects reproduction.

Mineralocorticoid and glucocorticoid receptor mRNA expression in the brain of translocated chukar (Alectoris chukar).

Although translocation is an important conservation tool in the effort to create self-sustaining wild populations of threatened species, avian translocations have a high failure rate and causes for failure are poorly understood. While "stress" is considered to play a major role in translocation failure, the physiological changes associated with chronic stress resulting from translocation have been investigated only recently. Translocation results in chronic stress-induced alterations of stress response physiology in the chukar (Alectoris chukar) and in the present study we tested the hypothesis that changes in the hypothalamic-pituitary-adrenal axis (HPA) are correlated with changes in the brain, specifically at the level of the glucocorticoid and mineralocorticoid receptors (GR and MR, respectively) in the hippocampus and hypothalamus. Our previous research has shown that there are apparent changes in GR and MR expression in the brain of experimentally chronically stressed European starlings (Sturnus vulgaris). In the present study however, translocation had no major detectable effect on levels of GR or MR mRNA expression in the hippocampus or hypothalamus of wild chukar suggesting that the observed dysregulation of the HPA axis by translocation may not be a result of such upstream changes.

Sex differences in brain aromatase activity: genomic and non-genomic controls.

Aromatization of testosterone into estradiol in the preoptic area plays a critical role in the activation of male copulation in quail and in many other vertebrate species. Aromatase expression in quail and in other birds is higher than in rodents and other mammals, which has facilitated the study of the controls and functions of this enzyme. Over relatively long time periods (days to months), brain aromatase activity (AA), and transcription are markedly (four- to sixfold) increased by genomic actions of sex steroids. Initial work indicated that the preoptic AA is higher in males than in females and it was hypothesized that this differential production of estrogen could be a critical factor responsible for the lack of behavioral activation in females. Subsequent studies revealed, however, that this enzymatic sex difference might contribute but is not sufficient to explain the sex difference in behavior. Studies of AA, immunoreactivity, and mRNA concentrations revealed that sex differences observed when measuring enzymatic activity are not necessarily observed when one measures mRNA concentrations. Discrepancies potentially reflect post-translational controls of the enzymatic activity. AA in quail brain homogenates is rapidly inhibited by phosphorylation processes. Similar rapid inhibitions occur in hypothalamic explants maintained in vitro and exposed to agents affecting intracellular calcium concentrations or to glutamate agonists. Rapid changes in AA have also been observed in vivo following sexual interactions or exposure to short-term restraint stress and these rapid changes in estrogen production modulate expression of male sexual behaviors. These data suggest that brain estrogens display most if not all characteristics of neuromodulators if not neurotransmitters. Many questions remain however concerning the mechanisms controlling these rapid changes in estrogen production and their behavioral significance.

Wild European starlings (Sturnus vulgaris) adjust to captivity with sustained sympathetic nervous system drive and a reduced fight-or-flight response.

Although research on wild species typically involves capture, handling, and some degree of captivity, few studies examine how these actions affect and/or alter the animal's underlying stress physiology. Furthermore, we poorly understand the immediate changes that occur as wild animals adjust to captive conditions. Most studies to date have investigated relatively long-term changes in the glucocorticoid response to an acute stressor, but immediate changes in the fight-or-flight response are relatively understudied in wild-caught species. In this study, we investigated changes to the cardiovascular stress response during the first 10 d of captivity of freshly captured wild European starlings (Sturnus vulgaris). We demonstrated that (1) baseline heart rate (HR) remains elevated for several days following transport into captivity, (2) the normal balance between sympathetic nervous system (SNS) and parasympathetic nervous system regulation of HR is disrupted, with the SNS exerting relatively greater control over baseline HR for the first days of captivity, and (3) the HR response to startle, a mild stressor, becomes significantly reduced compared to that of starlings maintained in captivity for several months and remains below the control response for at least 10 d. These data are the first to show that successive acute stressors and introduction to a captive setting significantly alter the physiology and responsiveness of the cardiovascular stress response system.

Combined effects of molt and chronic stress on heart rate, heart rate variability, and glucocorticoid physiology in European starlings.

Molt is an important life-history stage in avian species, but little is known about the effects of chronic stress during this period. Three weeks after the onset of molt, captive European starlings (Sturnus vulgaris) were exposed to 18 days of chronic stress, induced with four 30-minute randomized stressors presented daily. Birds showed no chronic-stress-induced changes in heart rate or heart rate variability when measured either during the middle of the day or at night. These data suggest that chronic stress did not alter the balance between sympathetic and parasympathetic nervous system regulation of cardiovascular function, which contrasts with data from an earlier study indicating that chronic stress profoundly alters cardiovascular function in non-molting starlings. Additionally, there was a significant increase in restraint-induced corticosterone secretion the first week of chronic stress that subsequently returned to pre-chronic-stress levels by the second week of exposure. The attenuated corticosterone response again contrasts with data from non-molting starlings that showed significant decreases in corticosterone responses. Consequently, the resistance to cardiovascular and corticosterone changes indicates that the physiological changes induced by chronic stress are greatly attenuated in molting birds. Overall, the data suggest that molt requires a degree of physiological stability that must be protected, so that if a bird is exposed to chronic stress during this life-history stage, molt takes priority.

The Reactive Scope Model - a new model integrating homeostasis, allostasis, and stress.

Allostasis, the concept of maintaining stability through change, has been proposed as a term and a model to replace the ambiguous term of stress, the concept of adequately or inadequately coping with threatening or unpredictable environmental stimuli. However, both the term allostasis and its underlying model have generated criticism. Here we propose the Reactive Scope Model, an alternate graphical model that builds on the strengths of allostasis and traditional concepts of stress yet addresses many of the criticisms. The basic model proposes divergent effects in four ranges for the concentrations or levels of various physiological mediators involved in responding to stress. (1) Predictive Homeostasis is the range encompassing circadian and seasonal variation - the concentrations/levels needed to respond to predictable environmental changes. (2) Reactive Homeostasis is the range of the mediator needed to respond to unpredictable or threatening environmental changes. Together, Predictive and Reactive Homeostasis comprise the normal reactive scope of the mediator for that individual. Concentrations/levels above the Reactive Homeostasis range is (3) Homeostatic Overload, and concentrations/levels below the Predictive Homeostasis range is (4) Homeostatic Failure. These two ranges represent concentrations/levels with pathological effects and are not compatible with long-term (Homeostatic Overload) or short-term (Homeostatic Failure) health. Wear and tear is the concept that there is a cost to maintaining physiological systems in the Reactive Homeostasis range, so that over time these systems gradually lose their ability to counteract threatening and unpredictable stimuli. Wear and tear can be modeled by a decrease in the threshold between Reactive Homeostasis and Homeostatic Overload, i.e. a decrease in reactive scope. This basic model can then be modified by altering the threshold between Reactive Homeostasis and Homeostatic Overload to help understand how an individual's response to environmental stressors can differ depending upon factors such as prior stressors, dominance status, and early life experience. We illustrate the benefits of the Reactive Scope Model and contrast it with the traditional model and with allostasis in the context of chronic malnutrition, changes in social status, and changes in stress responses due to early life experiences. The Reactive Scope Model, as an extension of allostasis, should be useful to both biomedical researchers studying laboratory animals and humans, as well as ecologists studying stress in free-living animals.

Stress and translocation: alterations in the stress physiology of translocated birds.

Translocation and reintroduction have become major conservation actions in attempts to create self-sustaining wild populations of threatened species. However, avian translocations have a high failure rate and causes for failure are poorly understood. While 'stress' is often cited as an important factor in translocation failure, empirical evidence of physiological stress is lacking. Here we show that experimental translocation leads to changes in the physiological stress response in chukar partridge, Alectoris chukar. We found that capture alone significantly decreased the acute glucocorticoid (corticosterone, CORT) response, but adding exposure to captivity and transport further altered the stress response axis (the hypothalamic-pituitary-adrenal axis) as evident from a decreased sensitivity of the negative feedback system. Animals that were exposed to the entire translocation procedure, in addition to the reduced acute stress response and disrupted negative feedback, had significantly lower baseline CORT concentrations and significantly reduced body weight. These data indicate that translocation alters stress physiology and that chronic stress is potentially a major factor in translocation failure. Under current practices, the restoration of threatened species through translocation may unwittingly depend on the success of chronically stressed individuals. This conclusion emphasizes the need for understanding and alleviating translocation-induced chronic stress in order to use most effectively this important conservation tool.

Heart rate and heart-rate variability responses to acute and chronic stress in a wild-caught passerine bird.

The cardiovascular-stress response has been studied extensively in laboratory animals but has been poorly studied in naturally selected species. We determined the relative roles of the sympathetic nervous system (SNS) and the parasympathetic nervous system (PNS) in regulating stress-induced changes in heart rate (HR) in wild-caught European starlings (Sturnus vulgaris). In both heart-rate variability (HRV) analysis and receptor blockade (atropine and propranolol) experiments, baseline HR was controlled predominantly by the PNS, whereas the increase in HR resulting from acute restraint stress was controlled predominantly by the SNS. These results indicate similar cardiac control of baseline and acute-stress-induced HR in wild-caught starlings and mammals. We further investigated HR responses during chronic stress. Driven primarily by changes in PNS regulation, baseline HR increased during the day but decreased at night. In addition, elevated HRs during acute restraint stress were attenuated throughout chronic stress and were accompanied by decreased HRV. This suggested that increased SNS drive elevated HR, but the attenuated HR response combined with resistance to the SNS blocker propranolol suggested that the sympathetic signal was less effective during chronic stress. Overall, chronic stress in wild-caught starlings elicited profound changes in cardiac function that were primarily regulated by changes in the PNS.

Initial transference of wild birds to captivity alters stress physiology.

Maintaining wild animals in captivity has long been used for conservation and research. While often suggested that captivity causes chronic stress, impacts on the underlying stress physiology are poorly understood. We used wild-caught chukar (Alectoris chukar) as a model avian species to assess how the initial 10 days of captivity alters the corticosterone (CORT) secretory pathway. In the first few days of captivity, birds lost weight, had lower hematocrit and demonstrated changes in CORT concentrations. Both baseline and restraint-stress-induced CORT concentrations decreased by days 3-5 of captivity and remained significantly lower throughout the 10 days although stress-induced concentrations began to recover by day 9. To delineate potential mechanisms underlying these CORT changes, we evaluated alterations to the hypothalamic-pituitary-adrenal (HPA) axis. Although chukar appear to be resistant to arginine vasotocin's (AVT) effects on CORT release, adrenocorticotropin hormone (ACTH) stimulated CORT release; however, ACTH stimulation did not differ during the 10 days of captivity. In contrast, negative feedback axis sensitivity, as determined by both dexamethasone suppression as well as endogenous negative feedback, decreased by day 5 but was regained by day 9. In addition, the combined stressors of capture and long distance transport eliminated the animals' ability to mount an acute CORT response on the day following the move. Therefore, introduction into captivity appeared to shift the chukar into a temporary state of chronic stress that began to recover within 9days. The duration of these alterations likely varies due to differences in capture techniques, transport distance, and species studied.

Captive European starlings (Sturnus vulgaris) in breeding condition show an increased cardiovascular stress response to intruders.

European starlings (Sturnus vulgaris) alter their physiology and behavior between seasons, becoming territorial during the spring/summer and flocking during the fall/winter. We used captive male starlings in breeding (photostimulated to 18L : 6D) and nonbreeding (11L : 13D) conditions to determine whether changing physiology and behavior alters their reaction to crowding. One or five intruders entered a resident's cage without human disturbance. A subcutaneous heart rate transmitter recorded cardiovascular output in residents. Corticosterone and testosterone were measured in plasma samples taken before and after the intrusion. While corticosterone concentrations did not change, heart rate changed significantly, indicating that these responses can be regulated independently. Long-day birds showed a significantly elevated heart rate response to the single-bird intrusion compared to short-day birds. Whereas five intruders elicited an identical peak response in both groups, long-day birds also demonstrated an equivalent response to one intruder. In addition, one intruder induced longer elevation in heart rate for long-day birds. Male starlings in breeding condition, therefore, demonstrate an increased sensitivity to additional conspecifics. This seasonal shift in response suggests that a higher tolerance for intrusion (i.e., considering a nearby starling as less stressful) may facilitate flocking behavior, while a lower tolerance may aid in territoriality.