Tactile experience induces c-fos expression in rat barrel cortex

Understanding gene expression that is responsive to sensory stimulation is central to elucidate molecular mechanisms underlying neuronal plasticity. In this study we demonstrate two new methods of stimulating whiskers that provide major sensory input to rat neocortex. In the first paradigm, animals were placed on the top of a cylinder and their vibrissae were brushed by hand. In the second paradigm, animals were placed for a brief period of time into a new, wired cage resulting in vibrissae stimulation when they explored the new environment. Both approaches induced c-Fos expression in barrel cortex corresponding to the stimulated vibrissae, especially in layer IV. Layers II/III and V/VI also showed c-Fos induction, but there were no detectable changes in layer VIb. The majority of c-Fos-expressing cells are probably not inhibitory neurons, because they do not show parvalbumin staining. Both paradigms, in contrast to the previous methods, are simple to use and do not require anesthesia, restraint of animals, or elaborate experimental setups.     

The adaptive response of adult rat Leydig cells to repeated immobilization stress: the role of protein kinase A and steroidogenic acute regulatory protein

The ability of immobilization stress (IMO) to decrease Leydig cell steroidogenesis and serum androgen concentration has been previously observed, but the possible mechanism(s) involved in the adaptation to prolonged or repeated stress have not been identified. In this study, we investigated whether the Leydig cells obtained from adult rats subjected to acute (15 min, 30 min or 2 h) and repeated (2 or 10 days, 2 h daily) IMO show adaptive mechanism(s) in response to stress-impaired steroidogenesis. The results showed that basal and human chorionic gonadotropin-stimulated cAMP production by Leydig cells isolated from rats exposed to both acute and repeated IMO was significantly reduced. Despite the reduced cAMP production, immunoblot analysis revealed increased immunoreactivity for both protein kinase A (PKA) and steroidogenic acute regulatory (StAR) protein in Leydig cells obtained from rats repeatedly exposed to IMO. Also, the phosphorylation and production of mature StAR protein was evident during exposure of rats to repeated IMO treatment. Treatment with cholesterol, the steroid substrate transported into mitochondria by StAR, significantly increased androgen and progesterone production by Leydig cells isolated from rats exposed to repeated IMO. In contrast, when other steroid substrates (22(R)-OH-cholesterol, pregnenolone, progesterone, Delta4-androstenedione) were present in the culture media, Leydig cell steroidogenesis was still reduced by IMO. Thus, PKA-mediated phosphorylation of StAR protein is an important mechanism in the adaptive response of Leydig cells to repeated IMO.     

Phosphorylated nerve growth factor-induced clone B (NGFI-B) translocates from the nucleus to mitochondria of stressed rat cardiomyocytes and induces apoptosis

Stress induces cardiac dysfunction and cardiomyocyte injury, and while current data indicate that mitochondria play a key role in this process, the mechanisms remain unknown. In this study, we found that in rats, restraint stress induced nerve growth factor-induced clone B (NGFI-B) translocation from the nucleus to mitochondria in cardiomyocytes. This translocation promoted cytochrome c release from mitochondria to the cytoplasm, which ultimately resulted in cardiomyocyte apoptosis. We also found that stress induced oversecretion of glucocorticoids and activated the protein kinase A (PKA) pathway in cardiomyocytes. Enhanced PKA activity increased NGFI-B serine phosphorylation, which caused NGFI-B to translocate from the nucleus to mitochondria. Moreover, a PKA peptide inhibitor blocked NGFI-B serine phosphorylation and translocation. Our data demonstrate that stress affects cardiomyocytes by inducing NGFI-B mitochondrial translocation via serine phosphorylation, which in turn initiates mitochondrial-mediated apoptosis.     

Olfactory stimulation induces filial preferences for huddling in rat pups.

Rat pups of all ages huddle with conspecifics, but the senosory control of contact behavior changes ontogenetically. Thermal cues control huddling until about Day 15, at which time species' odors become the dominant stimulus. The present experiments indicate that the filial response to conspecifics is dependent on olfactory experience. A synthetic chemical scent was added to the smells of the dam from Day 1 to Day 20 postpartum. Standardized videographic tests were used to assess the development of huddling preference. Preferences for nest-typical smells emerged by Day 15 in pups from both scented and nonscented litters. Pups from scented nests preferred to huddle with a scented stimulus rat, whereas control pups preferred a nonadulterated rat stimulus. Additional testing indicated that the affiliative preferences were specific to rearing odor and were not based on decreased aversion to test scents, or on disrupted olfactory discrimination. The ontogeny of species-typical contact behavior is discussed in terms of the induction of a perceptual preference that is based on early odor stimulation.     

Olfactory fear conditioning induces field potential potentiation in rat olfactory cortex and amygdala

The widely used Pavlovian fear-conditioning paradigms used for studying the neurobiology of learning and memory have mainly used auditory cues as conditioned stimuli (CS). The present work assessed the neural network involved in olfactory fear conditioning, using olfactory bulb stimulation-induced field potential signal (EFP) as a marker of plasticity in the olfactory pathway. Training consisted of a single training session including six pairings of an odor CS with a mild foot-shock unconditioned stimulus (US). Twenty-four hours later, the animals were tested for retention of the CS as assessed by the amount of freezing exhibited in the presence of the learned odor. Behavioral data showed that trained animals exhibited a significantly higher level of freezing in response to the CS than control animals. In the same animals, EFPs were recorded in parallel in the anterior piriform cortex (aPC), posterior piriform cortex (pPC), cortical nucleus of the amygdala (CoA), and basolateral nucleus of the amygdala (BLA) following electrical stimulation of the olfactory bulb. Specifically, EFPs recorded before (baseline) and after (during the retention test) training revealed that trained animals exhibited a lasting increase (present before and during presentation of the CS) in EFP amplitude in CoA, which is the first amygdaloid target of olfactory information. In addition, a transient increase was observed in pPC and BLA during presentation of the CS. These data indicate that the olfactory and auditory fear-conditioning neural networks have both similarities and differences, and suggest that the fear-related behaviors in each sensory system may have at least some distinct characteristics.     

Cytokine responses to LTP induction in the rat hippocampus: a comparison of in vitro and in vivo techniques

Because exogenous application of a number of cytokines and growth factors can alter synaptic properties, we sought to determine if endogenous cytokine expression is affected by neuronal activity. In addition, we examined whether cytokine expression is altered by the techniques used to stimulate and record from hippocampal neurons. Using semi-quantitative RNase protection and RT-PCR assays, we studied the expression of 18 cytokine, growth factor, and receptor genes in the hippocampus following the induction of Schaffer collateral-CA1 long-term potentiation (LTP). We found that various cytokines are dramatically induced following preparation of slices for in vitro recording and as a result of injury following acute electrode placement in vivo. These increases can be overcome in vivo, however, using permanent electrodes implanted three weeks prior to testing. Using this chronic preparation, we found that interleukin-6 (IL-6) mRNA was upregulated nearly 20-fold by LTP induction in vivo, marking the first demonstration of endogenous regulation of this cytokine in response to LTP. In situ hybridization for IL-6 revealed that upregulation is tightly localized near the site of stimulation and is detected only in non-neuronal cells, identified as GFAP+ astrocytes and GFAP− cells within proximal blood vessels. Coupled with previous results showing that exogenously applied IL-6 can prevent the induction of LTP, this finding suggests a mechanism by which the local release of a cytokine could regulate LTP at nearby sites.     

Stress applied during primary immunization affects the secondary humoral immune response in the rat: involvement of opioid peptides

The effect of unpredictable, inescapable and uncontrollable electric tail shocks (ES) on the humoral immune response to bovine serum albumin (BSA) was investigated in the rat. Contributions of the procedures that accompany shock delivery, such as witnessing the ES procedure (stress witnessing, SW) and exposure to the apparatus for shock delivery (apparatus control, AC) to the changes in specific immunity induced by ES were also tested. All procedures were applied during primary and/or secondary immunization. It was demonstrated that exposure to ES during primary immunization with BSA significantly suppressed specific anti-BSA antibody production after secondary and tertiary immunization with the same antigen. Exposure to the SW procedure during primary immunization with BSA enhanced the specific antibody level after secondary immunization, while exposure to the apparatus alone did not influence the development of either the primary or secondary humoral immune response to BSA. Both ES-induced suppression and SW-induced potentiation of the humoral immune response were partially inhibited by prior treatment with the opioid receptor antagonist naloxone. Additionally, treatments with the opioid peptides methionine- and leucine-enkephalin decreased anti-BSA antibody level, mimicking to some extent the effects of ES. It is suggested that ES and endogenous opioid peptides had long-term effects on humoral immunity through mechanisms involving immunologic memory.     

Enhanced proliferation of progenitor cells following long-term potentiation induction in the rat dentate gyrus

The dentate gyrus (DG) is among the few areas in the mammalian brain where production of new neurons continues in the adulthood. Although its functional significance is not completely understood, several lines of evidence suggest the role of DG neurogenesis in learning and memory. Considering that long-term potentiation (LTP) is a prime candidate for the process underlying hippocampal learning and memory, these results raise the possibility that LTP and neurogenesis are closely related. Here, we investigated whether or not LTP induction in the afferent pathway triggers enhanced proliferation of progenitor cells in the DG. LTP was induced by tetanic stimulation in perforant path-DG synapses in one hemisphere, and the number of newly generated progenitor (BrdU-labeled) cells in the DG was quantified. Compared with the control hemisphere (stimulated with low-frequency pulses), the LTP-induced hemisphere contained a significantly higher number of newly generated progenitor cells in the dorsal as well as ventral DG. When CPP, an NMDA receptor antagonist, was administered, tetanic stimulation neither induced LTP nor enhanced progenitor cell proliferation, indicating that NMDA receptor activation, rather than tetanic stimulation per se, is responsible for enhanced progenitor proliferation in the control animal. Our results show that tetanic stimulation of perforant path sufficient to induce LTP increases progenitor proliferation in adult DG in an NMDA receptor-dependent manner.     

The effect of stress and in vivo vasoactive intestinal peptide (VIP) treatment on the response of isolated rat aorta to norepinephrine, angiotensin II and vasopressin, and adventitial mast cells

The effects of cold-restraint stress, repeated over 3 days, and treatment of rats with vasoactive intestinal peptide (VIP) on the contractile responses of isolated aorta to vasoconstrictors, and on aortic adventitial mast cells were investigated. Stress significantly reduced the contractile response of rat aorta smooth muscle to norepinephrine (NE), angiotensin II (Ang II) and vasopressin (VP). Decreased sensitivity to NE, Ang II and VP may result from decreased receptor density, and affinity or reduced effector efficacy. Stress induced degranulation, decreased the number and changed the granular content of mast cells; all degranulated mast cells were stained with alcian blue, and the percentage of safranin staining cells was decreased. Given prior to stress, VIP reversed the reduced contractile responses and sensitivity of aorta to NE and Ang II but had no effect on VP subsensitivity. VIP also inhibited stress-induced degranulation of mast cells, and after VIP only alcian blue-stained mast cells were seen. When VIP was given to non-stressed rats, the contractile response of the aorta to NE, but not Ang II or VP, was increased compared with control. Mast cell count was decreased in the adventitia of non-stressed VIP treated rats. The results indicate that stress decreases the heparin content of mast cells and VIP has an additive effect. In conclusion, VIP modulates both stress-induced mast cell activity and reduced sensitivity of aorta smooth muscle to NE and Ang II. It can be suggested that VIP may moderate some effects of stress on vascular pathophysiology.     

Hormonal regulation of delta opioid receptor immunoreactivity in interneurons and pyramidal cells in the rat hippocampus

Clinical and preclinical studies indicate that women and men differ in relapse vulnerability to drug-seeking behavior during abstinence periods. As relapse is frequently triggered by exposure of the recovered addict to objects previously associated with drug use and the formation of these associations requires memory systems engaged by the hippocampal formation (HF), studies exploring ovarian hormone modulation of hippocampal function are warranted. Previous studies revealed that ovarian steroids alter endogenous opioid peptide levels and trafficking of mu opioid receptors in the HF, suggesting cooperative interaction between opioids and estrogens in modulating hippocampal excitability. However, whether ovarian steroids affect the levels or trafficking of delta opioid receptors (DORs) in the HF is unknown. Here, hippocampal sections of adult male and normal cycling female Sprague-Dawley rats were processed for quantitative immunoperoxidase light microscopy and dual label fluorescence or immunoelectron microscopy using antisera directed against the DOR and neuropeptide Y (NPY). Consistent with previous studies in males, DOR-immunoreactivity (-ir) localized to select interneurons and principal cells in the female HF. In comparison to males, females, regardless of estrous cycle phase, show reduced DOR-ir in the granule cell layer of the dentate gyrus and proestrus (high estrogen) females, in particular, display reduced DOR-ir in the CA1 pyramidal cell layer. Ultrastructural analysis of DOR-labeled profiles in CA1 revealed that while females generally show fewer DORs in the distal apical dendrites of pyramidal cells, proestrus females, in particular, exhibit DOR internalization and trafficking towards the soma. Dual label studies revealed that DORs are found in NPY-labeled interneurons in the hilus, CA3, and CA1. While DOR colocalization frequency in NPY-labeled neuron somata was similar between animals in the hilus, proestrus females had fewer NPY-labeled neurons that co-labeled with DOR in stratum oriens of CA1 and CA3 when compared to males. Ultrastructural analysis of NPY-labeled axon terminals within stratum radiatum of CA1 revealed that NPY-labeled axon terminals contain DORs that are frequently found at or near the plasma membrane. As no differences were noted by sex or estrous cycle phase, DOR activation on NPY-labeled axon terminals would inhibit GABA release probability equally in males and females. Taken together, these findings suggest that ovarian steroids can impact hippocampal function through direct effects on DOR levels and trafficking in principal cells and broad indirect effects through reductions in DOR-ir in NPY-labeled interneurons, particularly in CA1.     

Injection of 5,7-dihydroxytryptamine into the B3 raphe region of neonatal rat pups induces hyperalgesia but only slight alterations in ingestion-related behaviors.

The effects of intrabrainstem injections of the neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) into the B3 raphe region (nucleus raphe magnus and nucleus reticularis paragigantocellularis) on early ingestive behavior and nociception were assessed in Sprague-Dawley rat pups during the first postnatal week. Lesions resulted in a marked depletion of serotonin (5HT) in hindbrain without influencing 5HT levels in forebrain. Pretreatment with desipramine (DMI) resulted in a sparing of noradrenergic neurons from neurotoxic effects. The B3 lesion resulted in significant hyperalgesia as reflected by decreased latencies in tail flick testing. Although nipple attachment latencies in suckling tests were slightly increased by the lesion, no notable effects on mouthing or other ingestive-related behaviors were observed in testing conducted in an independent ingestion paradigm. These results suggest that whereas B3 serotonergic neurons may be functioning in an adult-typical manner to regulate analgesia during the early postnatal period, this raphe region may play only a slight role in the modulation of ingestion-related behaviors early in life.     

Ghrelin stimulates corticotropin-releasing factor and vasopressin gene expression in rat hypothalamic 4B cells

Corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) play a central role in regulating the stress response. In response to stress, CRF and AVP neurons in the hypothalamic paraventricular nucleus secrete the peptides to stimulate the release of adrenocorticotropic hormone from the anterior pituitary. Ghrelin, an endogenous ligand of the growth hormone-releasing peptide receptors (GHSR), has been shown to stimulate the release of CRF and AVP by rat hypothalamic explants. However, little is known about the ability of the ghrelin signaling pathways to activate the CRF and AVP genes in the hypothalamus. In the present study, we examined the direct effect of ghrelin on CRF and AVP gene expression in hypothalamic 4B cells, which show the characteristics of the hypothalamic parvocellular paraventricular nucleus neurons. Cells were transfected with CRF or AVP promoter to examine the activity of each promoter. Ghrelin stimulated the promoter activities and mRNA levels for both CRF and AVP. The involvement of a protein kinase pathway was examined using inhibitors. Protein kinase A and phospholipase C pathways were shown to be involved in ghrelin-induced increases in both CRF and AVP promoter activities. GHSR type 1a (GHSR1a) mRNA levels were also increased by ghrelin, and these ghrelin-induced levels were suppressed by a GHSR1a antagonist. Thus, ghrelin-dependent pathways are involved in the regulation of CRF and AVP gene expression in the hypothalamus: ghrelin, an orexigenic hormone, stimulates CRF, an anorexigenic/anxiogenic factor in the hypothalamus, resulting in hypothalamic-pituitary-adrenal axis activation to stimulate the release of glucocorticoids.     

损毁大鼠双侧内嗅皮质对束缚应激反应的影响

采用大鼠急性束缚应激动物模型,用鹅羔氨酸损毁大鼠双侧内嗅皮质来建立内嗅皮质损伤模型,观察束缚应激1小时后下丘脑室旁核快反应基因c-Fos表达情况以及应激后血浆促肾上腺皮质激素含量的动态变化,并与对照组相比较,探讨内嗅皮质与束缚应激反应的关系。结果表明,损毁内嗅皮质,明显抑制了应激诱导的下丘脑室旁核c-Fos的表达和血浆促肾上腺皮质激素含量的上升。结果提示,内嗅皮质参与调节束缚应激反应时HPA轴功能。     

Levels of Meaning in Family Stress Theory

Major stressful life events, particularly those that have chronic hardships, create a crisis for families that often leads to reorganization in the family's style of functioning. A major factor in this reorganization is the meaning the family gives to the stressful event. Often the meaning extends beyond the event itself and leads to a changed view of the family system and even to a changed view of the world. Building on other family stress models, we elaborate the family's definition of the stressor into three levels of family meanings: (1) situational meanings, (2) family identity, and (3) family world view. Examples from clinical work and studies of families adapting to chronic illness are used to illustrate the relationship between these three levels of meaning, particularly as they change in response to crisis. Implications for clinical and empirical work are discussed.     

A novel in vivo measure of cyberaggression

Cyberaggression (CA), or the use of information communication technologies to inflict harm on others, is an emerging public health crisis. Unfortunately, our current ability to assess CA in a research context remains limited, curtailing efforts to address this important issue. We sought to fill this gap in the literature by developing an adapted “chat” version of the Taylor aggression paradigm (TAP) that would more closely resemble a social gaming format (hereafter referred to as the TAP-Chat). In the TAP-Chat, participants have a chat function available to communicate with their (fictitious) co-player. Following loss trials in a competitive reaction time task, they receive a “mean chat” from their co-player. Participant messages to their (fictitious) co-player are then coded for aggressive content by a team of trained research assistants, and via automated linguistic analysis software (Linguistic Inquiry and Word Count). The current study evaluated the predictive utility of the TAP-Chat task in independent discovery and replication samples (N = 843 and N = 350, respectively). Participants’ publicly available tweets served as an important external criterion variable, along with a handful of self-report questionnaires assessing CA and related constructs. Analyses suggest that, although it can be completed in ∼13 min, the TAP-Chat predicts CA on Twitter and, to a lesser extent, as reported on questionnaires. Although there are still several issues to address, it is our hope that the research community will benefit from this straightforward behavioral assessment of CA.     

Stress Reduction for Family Caregivers in Chronic Mental Illness: Implications of a Work Stress Management Perspective

In several studies involving a total of 291 family caregivers for schizophrenia sufferers, the stressors that arise from caregiving were identified. Also identified were the outcomes for caregivers, which often include psychological distress. Caregivers develop various stress-reduction techniques, but this article explores the utility of applying the principles of work stress management to caregiver well-being. An organizational psychology perspective suggests that a comprehensive focus must include not only how individuals can learn to manage the emotional demands of their work, but also how the work of caregiving can be made less stressful for them. Suggestions from a work stress management perspective highlight the possible contributions of worker participation in policy formulation and a collaborative relationship between family and professional caregivers. Potentially fruitful research directions are noted.     

Predictors of Stress Generation in Adolescents in Mainland China

The current longitudinal study examined whether the personality vulnerabilities of self-criticism and dependency prospectively predicted stress generation in Chinese adolescents. Participants included 1,116 adolescents (588 girls and 528 boys), aged 15 to 18 years from rural, urban and ultra-urban mainland China. Participants completed self-report measures of personality, depressive and anxious symptoms and participated in a clinical interview assessing lifetime history of depression. The occurrence of negative life events was measured using a contextual-threat interview every 6-months for a total period of 18-months. Logistic regression analyses showed that after controlling for past depressive episodes and current depressive and anxious symptoms, self-criticism was prospectively associated with the occurrence of interpersonal stress generation, but not noninterpersonal stress generation. Dependency also predicted interpersonal stress generation, although only in girls and not boys. In line with previous Western findings, girls reported more interpersonal stress generation. Analyses across 3 levels of urbanization revealed several significant differences including higher reported interpersonal stress generation in urban girls than urban boys and overall higher levels of negative life events in ultra-urban youth. In sum, findings from the current study suggest that the stress generation process may be generalizable to Chinese youth.     

Psychosocial Predictors of Acculturative Stress in Mexican Immigrants

Psychosocial predictors of acculturative stress were examined in a sample of adult Mexican immigrants in Los Angeles. Bivariate and multivariate analyses revealed that family dysfunction, geographical separation from family, nonpositive expectations for the future, and low income levels were significantly related to elevated levels of acculturative stress. The findings suggest that family closeness, hopefulness for the future, and financial resources may provide a buffer against acculturative Stressors experienced by migrating individuals. The findings highlight the importance of using culturally relevant clinical methods when assessing and treating immigrants and acculturating individuals.     

Effects of acute, repeated and chronic variable stress on in vivo tyrosine hydroxylase activity and on alpha(2)-adrenoceptor sensitivity in the rat brain

We assessed the effects of a single tail pinch and two chronic stress regimes, repeated and variable, on in vivo tyrosine hydroxylase activity and on alpha2-adrenoceptor sensitivity in two brain regions. After administering a 3,4-dihydroxyphenylalanine (DOPA) decarboxylase inhibitor, tyrosine hydroxylase activity, measured as the accumulation of DOPA, and noradrenaline (NA) content were determined by using high-performance liquid chromatography. A single tail pinch for 5 min induced an enhancement of DOPA content in hippocampus (28%) and hypothalamus (67%) which was still present 24 h later. This increase could account for the lack of changes in NA content in both regions after the application of this stressor. However, tyrosine hydroxylase activity was unmodified 24 h after exposure to both repeated (5 min of tail pinch, twice daily, for 14 days) and chronic variable stress (one of 5 different stressors, once daily, for 14 days) although there was an enhancement of NA levels in hippocampus (45 and 54%, respectively) and hypothalamus (24.5 and 36%, respectively). The sensitivity of the alpha2-adrenoceptors which regulate [3H]-NA release in hippocampal and hypothalamic synaptosomes was not modified by the acute or chronic stress protocols assayed. The results show that both paradigms of chronic stress had similar effects on the noradrenergic indices evaluated.