Elicit Stress Responses Research Articles

Rumor Has It…: Relay Communication of Stress Cues in Plants

Recent evidence demonstrates that plants are able not only to perceive and adaptively respond to external information but also to anticipate forthcoming hazards and stresses. Here, we tested the hypothesis that unstressed plants are able to respond to stress cues emitted from their abiotically-stressed neighbors and in turn induce stress responses in additional unstressed plants located further away from the stressed plants. Pisum sativum plants were subjected to drought while neighboring rows of five unstressed plants on both sides, with which they could exchange different cue combinations. On one side, the stressed plant and its unstressed neighbors did not share their rooting volumes (UNSHARED) and thus were limited to shoot communication. On its other side, the stressed plant shared one of its rooting volumes with its nearest unstressed neighbor and all plants shared their rooting volumes with their immediate neighbors (SHARED), allowing both root and shoot communication. Fifteen minutes following drought induction, significant stomatal closure was observed in both the stressed plants and their nearest unstressed SHARED neighbors, and within one hour, all SHARED neighbors closed their stomata. Stomatal closure was not observed in the UNSHARED neighbors. The results demonstrate that unstressed plants are able to perceive and respond to stress cues emitted by the roots of their drought-stressed neighbors and, via ‘relay cuing’, elicit stress responses in further unstressed plants. Further work is underway to study the underlying mechanisms of this new mode of plant communication and its possible adaptive implications for the anticipation of forthcoming abiotic stresses by plants.

Vasopressin needs an audience: Neuropeptide elicited stress responses are contingent upon perceived social evaluative threats

The nonapeptide arginine vasopressin (AVP) plays an important role in hypothalamus–pituitary–adrenal axis regulation and also functions as a social hormone in a wide variety of species, from voles to humans. In the current report we use a variety of stress inducing tasks, including the Trier Social Stress Test (TSST) and intranasal administration of AVP to show that intranasal administration of this neuropeptide leads to a significant increase in salivary cortisol and pulse rate, specifically in conditions where subjects perform tasks in the presence of a social evaluative threat (task performance could be negatively judged by others). In contrast, in conditions without a social evaluative threat (no task condition, modified TSST without audience and bike ergometry), subjects receiving AVP did not differ from subjects receiving placebo. Thus exogenous AVP's influence is contingent upon a circumscribed set of initial conditions that constitute a direct threat to the maintenance of our social selves. Stress evoked by social threat is an integral part of social life and is related to self-esteem and in extreme forms, to poor mental health (e.g., social phobia). Our findings suggest that AVP is a key component in the circuit that interlaces stress and social threat and findings offer inroads to our understanding of individual differences in sociability and in stress response elicited in threatening social situations.

DNA Damage-Mediated Induction of a Chemoresistant Niche

While numerous cell-intrinsic processes are known to play decisive roles in chemotherapeutic response, relatively little is known about the impact of the tumor microenvironment on therapeutic outcome. Here, we use a well-established mouse model of Burkitt's lymphoma to show that paracrine factors in the tumor microenvironment modulate lymphoma cell survival following the administration of genotoxic chemotherapy. Specifically, IL-6 and Timp-1 are released in the thymus in response to DNA damage, creating a "chemo-resistant niche" that promotes the survival of a minimal residual tumor burden and serves as a reservoir for eventual tumor relapse. Notably, IL-6 is released acutely from thymic endothelial cells in a p38-dependent manner following genotoxic stress, and this acute secretory response precedes the gradual induction of senescence in tumor-associated stromal cells. Thus, conventional chemotherapies can induce tumor regression while simultaneously eliciting stress responses that protect subsets of tumor cells in select anatomical locations from drug action.

Using stressful films to analyze risk factors for PTSD in analogue experimental studies – which film works best?

To understand mental disorders, analogue paradigms provide an indispensable contribution. In posttraumatic stress disorder (PTSD), the stressful film paradigm is a frequently used analogue approach: Films depicting traumatic events are shown to non-clinical participants in order to elicit stress responses analogue to responses to traumatic events in real life. Previous studies used a large variety of films, which is problematic with regard to the comparability of results. The main goal of this study was to identify a film clip that (a) consistently provokes stress reactions and (b) provokes reactions that are as similar as possible to traumatic stress. We randomly exposed 105 male and female participants to one of four stressful films, differing, e.g., in content and origin. Intrusive memories of the film, reported immediately after the film and during a diary phase of three days, as well as distress, heart rate, and several mood states were measured. A film clip depicting rape elicited the most consistent reactions that were characterized by a higher heart rate, more distress and more intrusive memories, compared to the other three clips. Intrusive memories across all films were especially related to an increase in heart rate and disgust in response to the film.

Actein activates stress‐ and statin‐associated responses and is bioavailable in Sprague‐Dawley rats

The purpose of this study was to assess in rats the pharmacological parameters and effects on gene expression in the liver of the triterpene glycoside actein. Actein, an active component from the herb black cohosh, has been shown to inhibit the proliferation of human breast cancer cells. To conduct our assessment, we determined the molecular effects of actein on livers from Sprague-Dawley rats treated with actein at 35.7 mg/kg for 6 and 24 h. Chemogenomic analyses indicated that actein elicited stress and statin-associated responses in the liver; actein altered expression of cholesterol and fatty acid biosynthetic genes, p53 pathway genes, CCND1 and ID3. Real-time RT-PCR validated that actein induced three time-dependent patterns of gene expression in the liver: (i) a decrease followed by a significant increase of HMGCS1, HMGCR, HSD17B7, NQO1, S100A9; (ii) a progressive increase of BZRP and CYP7A1 and (iii) a significant increase followed by a decrease of CCND1 and ID3. Consistent with actein's statin- and stress-associated responses, actein reduced free fatty acid and cholesterol content in the liver by 0.6-fold at 24 h and inhibited the growth of human HepG2 liver cancer cells. To determine the bioavailability of actein, we collected serum samples for pharmacokinetic analysis at various times up to 24 h. The serum level of actein peaked at 2.4 microg/mL at 6 h. Actein's ability to alter pathways involved in lipid disorders and carcinogenesis may make it a new agent for preventing and treating these major disorders.

Amiodarone induces stress responses and calcium flux mediated by the cell wall inSaccharomyces cerevisiae

We used a proteomic approach to study effects of amiodarone on cells of the yeast Saccharomyces cerevisiae. Amiodarone has been shown to have antifungal activity in vitro and causes a massive increase in cytoplasmic calcium levels ([Ca2+]cyt). Proteomic analysis of cells exposed to amiodarone show that this drug elicits stress responses and points to involvement of proteins associated with the cell wall. We tested several of those proteins for involvement in the Ca2+ flux. In particular, the amiodarone-induced Ca2+ flux was decreased in bgl2Delta cells, which have altered levels of beta-glucan and chitin. The involvement of the cell wall in the Ca2+ flux induced by amiodarone treatment was tested by addition of yeast cell-wall components. While mannan inhibited the rise in [Ca2+]cyt, beta-glucan potentiated the Ca2+ flux by 4.5-fold, providing evidence that the cell wall is directly involved in controlling this Ca2+ flux. This conclusion is corroborated by the inhibition of the Ca2+ flux by calcofluor, which is known to bind to cell-wall chitin and inhibit cell growth. Zymolyase treatment altered the kinetics of amiodarone-induced calcium flux and uncoupled the inhibitory effect of calcofluor. These effects demonstrate that the cell-wall beta-glucan regulates calcium flux elicited by amiodarone.

Thrombospondin-1 and CD47 Limit Cell and Tissue Survival of Radiation Injury

Radiation, a primary mode of cancer therapy, acutely damages cellular macromolecules and DNA and elicits stress responses that lead to cell death. The known cytoprotective activity of nitric oxide (NO) is blocked by thrombospondin-1, a potent antagonist of NO/cGMP signaling in ischemic soft tissues, suggesting that thrombospondin-1 signaling via its receptor CD47 could correspondingly increase radiosensitivity. We show here that soft tissues in thrombospondin-1-null mice are remarkably resistant to radiation injury. Twelve hours after 25-Gy hindlimb irradiation, thrombospondin-1-null mice showed significantly less cell death in both muscle and bone marrow. Two months after irradiation, skin and muscle units in null mice showed minimal histological evidence of radiation injury and near full retention of mitochondrial function. Additionally, both tissue perfusion and acute vascular responses to NO were preserved in irradiated thrombospondin-1-null hindlimbs. The role of thrombospondin-1 in radiosensitization is specific because thrombospondin-2-null mice were not protected. However, mice lacking CD47 showed radioresistance similar to thrombospondin-1-null mice. Both thrombospondin-1- and CD47-dependent radiosensitization is cell autonomous because vascular cells isolated from the respective null mice showed dramatically increased survival and improved proliferative capacity after irradiation in vitro. Therefore, thrombospondin-1/CD47 antagonists may have selective radioprotective activity for normal tissues.

Comparative Analyses of Abscisic Acid Responses in Plants

The phytohormone abscisic acid (ABA) is a key mediator of various physiological and developmental processes in plants. It plays a major role in eliciting stress responses via a complex network of signaling pathways. We are using a rice suspension cell culture system and monitoring the effects of ABA exposure. These changes are being followed at the levels of both protein and metabolites. Proteomic profiles are monitored using iTRAQ peptide tagging and DIGE, complementary approaches that provide information about abundance and changes in post translational modifications. Metabolomic analysis is also being conducted simultaneously using metabolite extracts to monitor carbohydrate, organic acid and amino acid profiles through GC-MS technology. A quantitative PCR analysis on a set of known ABA response regulators was performed in parallel to validate ABA responses. These investigations are beginning to allow an understanding of relative protein abundance changes, metabolite responses and provide insights into ABA effects at the molecular and cellular levels.

Behavioral, metabolic, and molecular stress responses of marine bivalve Mytilus galloprovincialis during long-term acclimation at increasing ambient temperature

The present study aimed to determine the thermal response of the Mediterranean mussel Mytilus galloprovincialis by integrating information from various levels of biological organization including behavior, metabolic adjustments, heat shock protein expression, and protein kinase activity. Behavioral responses were determined by examining the effect of warming on valve closure and opening. Metabolic impacts were assessed by examining the activity of the key glycolytic enzyme pyruvate kinase (PK). Molecular responses were addressed through the expression of Hsp70 and Hsp90 and the phosphorylation of stress-activated protein kinases, p38 mitogen-activated protein kinase (p38 MAPK) and cJun-N-terminal kinases (JNKs). Mussels increased the duration of valve closure by about sixfold when acclimated to 24 degrees C rather than to 17 degrees C. As indicated by the activity of PK, such behavior caused metabolic depression and probably a shift from aerobic to anaerobic metabolism. Acclimation to temperatures higher than 24 degrees C caused an increase in mortality and induced the expression of Hsp72. Increased phosphorylation of p38 MAPK and JNKs indicated activation of MAPK signaling cascades. The potential involvement of MAPKs in the induction of Hsp genes in the tissues of M. galloprovincialis is discussed. In conclusion, it seems that M. galloprovincialis lives close to its acclimation limits and incipient lethal temperature and that a small degree of warming will elicit stress responses at whole organism and molecular levels.

Ferret odor as a processive stress model in rats: neurochemical, behavioral, and endocrine evidence.

Predator odors have been shown to elicit stress responses in rats. The present studies assessed the use of domestic ferret odor as a processive stress model. Plasma corticosterone and adrenocorticotropin hormone levels were higher after 30 min of exposure to ferret odor (fur/skin) but not control odors, ferret feces, urine, or anal gland secretions. Behavioral differences were also found between ferret and the control odors as tested in a defensive withdrawal paradigm. In addition, c-fos messenger RNA expression in several brain areas previously associated with processive stress was significantly higher in ferret odor-exposed rat brains than in control odor-exposed brains. These results suggest that ferret odor produces a reliable unconditioned stress response and may be useful as a processive stress model.

The pattern of brain c- fos mRNA induced by a component of fox odor, 2,5-dihydro-2,4,5-Trimethylthiazoline (TMT), in rats, suggests both systemic and processive stress characteristics

Predators to rodents and their associated odors are increasingly chosen to study the neural mechanisms of stress and anxiety. Specifically, predatory odors are believed to elicit responses based on the perceived threat (psychological or processive), rather than to any direct systemic effects (pain, blood loss, infection, etc.) of the stimulus, which are mediated by distinct neural pathways. The hypothesis that a chemical component from fox feces, 2,5-dihydro-2,4,5-trimethylthiazoline (TMT), elicits stress responses by specific activation of processive neural pathways was tested. Different amounts of TMT (range: 0–600 μmol) or the control odor butyric acid (0-1200 μmol) were presented to male Sprague–Dawley rats for 30 min. Immediately after odor presentation, rats were sacrificed, blood levels of adrenocorticotropic hormone (ACTH) and corticosterone were measured, and brains were rapidly harvested to measure regional brain c- fos mRNA induction by in situ hybridization. Presentation of TMT (≥75 μmol), but not butyric acid (up to 1200 μmol), significantly increased ACTH and corticosterone release. TMT presentation, especially with amounts (≥75 μmol) producing endocrine activation, induced c- fos mRNA in several brain areas, including the olfactory bulb, lateral septal nucleus, septohypothalamic nucleus, anteromedial and oval nuclei of the bed nucleus of the stria terminalis, the central nucleus of the amygdala, the anteroventral, anterodorsal, and medial preoptic nuclei, the anterior, dorsomedial, lateral, supramammillary, dorsal premammillary and paraventricular hypothalamic nuclei, the external lateral parabrachial nucleus, the locus coeruleus, and the nucleus of the solitary tract. Interestingly, these brain regions represent a mix of regional c- fos mRNA induction pattern not reported previously with any other single stressor. These results suggest that TMT elicits stress responses through a relatively unique and complex mix of brain regions associated with both processive and systemic neural pathways, unlike those seen in response to cat odors.

AUDITORY INFORMATION PROCESSING IS ALTERED IN NOVELTY STRESS CONDITIONS: FIRST SESSION EFFECTS IN AUDITORY-EVOKED POTENTIALS

Novelty conditions may elicit stress responses. First session effects are systematic changes in physiological parameters, resulting from the interference of physiological processes with novelty stress. Along with endocrinological changes, these stress responses may be accompanied by alterations of sensory and attentional processes. The present study examines the impact of novelty conditions on event-related potential indicators of auditory information processing and on cortisol. Twenty-two healthy subjects participated in a series of experimental sessions. Auditory-evoked potentials were recorded, and the plasma cortisol levels were determined. The first session causes an activation of the hypothalamo-hypopituitary-adrenal axis. The auditory-evoked potentials show an additional slow negative potential component in the novelty condition. This potential component is maximal at fronto-central electrode sites and reaches its peak at about 240 ms after stimulus onset. Similarities with the processing negativity suggest alterations in attention-related auditory information-processing in the novelty condition encountered in the first session

Anthropogenic Sound: Effects on the Behavior and Physiology of Fishes

Anthropogenic sound in the marine environment continues to increase. Sound sources range from increased vessel traffic to transient but intense sounds such as those produced by seismic air guns, pile driving, or some sonars. While most interest in anthropogenic sounds has focused on marine mammals, there is an increasing concern regarding the impact of such sounds on fishes and marine invertebrates. Since the inner ear hearing receptors of fishes are similar to those of marine mammals, any effects seen on the hearing receptors of marine mammals may also be found in fishes and vice versa. Despite increasing interest in the effects of sounds on fishes, this issue has only been addressed on the most limited scale. Here we review the current literature in this area. It has been reported that high sound levels can damage the inner ear sensory cells, produce hearing loss (threshold shifts), elicit stress responses, and alter the behavior of fishes. At least in terms of hearing loss, these effects are modulated by exposure sound level and duration. The effects of various types of sound (e.g., impulsive vs. continuous) and long-term impacts of how anthropogenic sounds affect the behavior and ecology of fishes need exploration in the future.

Neuroimmune Stress Responses: Reciprocal Connections Between the Hypothalamus and the Brainstem

Hypothalamic nuclei, particularly the paraventricular nuclei (PVN), are important brain sites responsible for eliciting stress responses following a systemic immune challenge. The activation of PVN cells by a systemic immune challenge is critically dependent on the integrity of inputs from brainstem cells situated in the nucleus tractus solitarius (NTS) and ventrolateral medulla (VLM). Interestingly, a descending pathway from the PVN to the brainstem, recruited by systemic immune challenge, might also exist. It is well documented that PVN neurons innervate the NTS and VLM and recent evidence from our laboratory shows that lesions of the PVN reduce brainstem cell responses elicited by a systemic bolus of the proinflammatory cytokine interleukin-1 g (IL-1 g ). Although a number of different PVN divisions are candidates for the source of inputs to the brainstem, we have demonstrated that the majority of descending PVN projections recruited by systemic IL-1 g arise from cell bodies localized in the medial and lateral parvocellular PVN. These findings suggest that central nervous system responses to an immune challenge are likely to involve complex reciprocal connections between the PVN and the brainstem, whereby brainstem cell populations could essentially act as integratory sites for descending and ascending immune signals. For instance, these brainstem pathways may have significant implications not only for the regulation of central hypothalamic and extra-hypothalamic targets but also the autonomic nervous system.

Cardiorespiratory responses of white sturgeon to environmental hypercapnia.

Cardioventilatory variables and blood-gas, acid-base status were measured in cannulated white sturgeon (Acipenser transmontanus) maintained at 19 degrees C during normocapnic and hypercapnic (Pw(CO(2)) approximately 20 Torr) water conditions and after the injection of adrenergic analogs. Hypercapnia produced significant increases in arterial PCO(2), ventilatory frequency, and plasma concentration of cortisol and epinephrine, and it produced significant decreases in arterial pH and plasma concentration of glucose but no change in arterial PO(2), hematocrit, and concentration of lactate or norepinephrine. Hypercapnia significantly increased cardiac output (Q) by 22%, mean arterial pressure (MAP) by 8%, and heart rate (HR) by 8%. However, gut blood flow (GBF) remained constant. In normocapnic fish, phenylephrine significantly constricted the splanchnic circulation, whereas isoproterenol significantly increased Q and produced a systemic vasodilation. During hypercapnia, propranolol significantly decreased Q, GBF, MAP, and HR, whereas phentolamine significantly decreased MAP and increased GBF. These changes suggest that cardiovascular function in the white sturgeon is sensitive to both alpha- and beta-adrenergic modulation. We found microspheres to be unreliable in predicting GBF on the basis of our comparisons with simultaneous direct measurements of GBF. Overall, our results demonstrate that environmental hypercapnia (e.g., as is experienced in high-intensity culture situations) elicits stress responses in white sturgeon that significantly elevate steady-state cardiovascular and ventilatory activity levels.

Seasonal variations of thyroid hormone levels in wild fish

Variations in plasma thyroxine (T4) and triiodothyronine (T3) contents of pike, pikeperch, perch, bream and chub from a large reservoir (Pareloup, South of France) were investigated before and during the progressive emptying of the reservoir. Fish were caught by gill nets, except at the last time of emptying when controls were collected with landing nets. Both iodothyronine levels showed desynchronized seasonal cycles resulting in higher T3/T4 ratios in fish stressed by gill-netting in early summer. The aforementioned results principally emphasized the prevailing influence of the technique of catching on eliciting stress responses and subsequent thyroidal impairment in all five species. However, the much higher amounts of T4 and/or T3 observed in specimens caught in the later stages of draining gave evidence to suggest that the progressive alteration in habitat might also have indirectly affected the thyroidal status of these fish, this latter result leading to the consideration of fish plasma thyroid hormones as possible endocrine biomarkers of chronic perturbation.

A leucine-rich nuclear export signal in the p53 tetramerization domain: regulation of subcellular localization and p53 activity by NES masking.

Appropriate subcellular localization is crucial for regulating p53 function. We show that p53 export is mediated by a highly conserved leucine-rich nuclear export signal (NES) located in its tetramerization domain. Mutation of NES residues prevented p53 export and hampered tetramer formation. Although the p53-binding protein MDM2 has an NES and has been proposed to mediate p53 export, we show that the intrinsic p53 NES is both necessary and sufficient for export. This report also demonstrates that the cytoplasmic localization of p53 in neuroblastoma cells is due to its hyperactive nuclear export: p53 in these cells can be trapped in the nucleus by the export-inhibiting drug leptomycin B or by binding a p53-tetramerization domain peptide that masks the NES. We propose a model in which regulated p53 tetramerization occludes its NES, thereby ensuring nuclear retention of the DNA-binding form. We suggest that attenuation of p53 function involves the conversion of tetramers into monomers or dimers, in which the NES is exposed to the proteins which mediate their export to the cytoplasm.

Confined compression of articular cartilage: Linearity in ramp and sinusoidal tests and the importance of interdigitation and incomplete confinement

Experimental and theoretical methods were used to investigate the linearity of the stress response of articular cartilage to ramp and sinusoidal tests in confined compression, as well as the role of cartilage-porous platen and lateral confinement boundary conditions in determining material responses. With respect to linearity, we posed the question as to whether the elicited stress responses to ramp compression, ramp release and sinusoidal tests were similar. With respect to boundary conditions we inquired as to the necessity of specifying a detailed interdigitating contact with the porous filter and of specifying the level of confinement present at the lateral edge of the disk. We found that the stress responses to the three types of tests were dissimilar, with ramp compression the only test exhibiting linear behavior. Ramp release from a static compression offset was non-linear in a manner such that the cartilage maintained a compressive stress higher than expected by a linear theory. Sinusoidal compression also displayed a non-linear response consistent with the presence of a release phase in each cycle. The actual boundary conditions present at the cartilage/porous-filter interface were visualized histologically. Areas (tens of microns) of cartilage in contact with the metal of the filter were interspersed with areas expanded into the pores of the filter. Finite-element analysis incorporating this information suggested that precise specification of this interface and of the level of the extent of lateral confinement would be necessary for the estimation of material properties, such as the hydraulic permeability, from these tests. The trends of the linearity studies did not appear to be significantly affected by the problems posed by these difficult to quantitate boundary conditions. The non-linear cartilage response to release and sinusoidal displacements therefore appear to be physiologically interesting. The maintained, that is higher than would be linear, compressive stress observed during release may be a beneficial adaptation to repeated loading where temporal variations in tissue stresses would be minimized.

The Stress Tolerance Test.

The Stress Tolerance Test combines principles and concepts from the Rorschach and TAT methods. Subjects write answers to standard questions about their impressions of slide-projected pictures of dramatic combat scenes designed to elicit stress responses among military personnel. Developed during World War II, this instrument has had little subsequent use. Its potential for yielding useful assessment information is illustrated in hopes of reawakening interest in it.