Modulation of PARP-1 Activity in a Broad Time Window Attenuates Memorizing Fear.

The amygdala plays a critical role in the acquisition and consolidation of fear-related memories. Recent studies have demonstrated that ADP-ribosylation of histones, accelerated by PARPs, affects the chromatin structure and the binding of chromatin remodeling complexes with transcription factors. Inhibition of PARP-1 activity during the labile phase of re-consolidation may erase memory. Accordingly, we investigated the possibility of interfering with fear conditioning by PARP-1 inhibition. Herein, we demonstrate that injection of PARP-1 inhibitors, specifically into the CeA or i.p., in different time windows post-retrieval, attenuates freezing behavior. Moreover, the association of memory with pharmacokinetic timing of PARP inhibitor arrival to the brain enabled/achieved attenuation of a specific cue-associated memory of fear but did not hinder other memories (even traumatic events) associated with other cues. Our results suggest using PARP-1 inhibitors as a new avenue for future treatment of PTSD by disrupting specific traumatic memories in a broad time window, even long after the traumatic event. The safety of using these PARP inhibitors, that is, not interfering with other natural memories, is an added value.

Phosphodiesterase-2 inhibitor reverses post-traumatic stress induced fear memory deficits and behavioral changes via cAMP/cGMP pathway.

Phosphodiesterase 2 is one of the phosphodiesterase (PDEs) family members that regulate cyclic nucleotide (namely cAMP and cGMP) concentrations. The present study determined whether PDE2 inhibition could rescue post-traumatic stress disorder (PTSD)-like symptoms. Mice were subjected to single prolonged stress (SPS) and treated with selective PDE2 inhibitor Bay 60-7550 (0.3, 1, or 3 mg/kg, i.p.). The behavioral tests such as forced swimming, sucrose preference test, open field, elevated plus maze, and contextual fear paradigm were conducted to determine the effects of Bay 60-7550 on SPS-induced depression- and anxiety-like behavior and fear memory deficits. The results suggested that Bay 60-7550 reversed SPS-induced depression- and anxiety-like behavior and fear memory deficits. Moreover, Bay 60-7550 prevented SPS-induced changes in the adrenal gland index, synaptic proteins synaptophysin and PSD95 expression, PKA, PKG, pCREB, and BDNF levels in the hippocampus and amygdala. These effects were completely prevented by PKG inhibitor KT5823. While PKA inhibitor H89 also prevented Bay 60-7550-induced pCREB and BDNF expression, but only partially prevented the effects on PSD95 expression in the hippocampus. These findings suggest that Bay 60-7550 protects mice against PTSD-like stress induced traumatic injury by activation of cGMP- or cAMP-related neuroprotective molecules, such as synaptic proteins, pCREB and BDNF.

The Role of Impulsivity in the Association Between Posttraumatic Stress Disorder Symptom Severity and Substance Use in Male Military Veterans.

High rates of posttraumatic stress disorder (PTSD) and comorbid substance use disorder (SUD) are prevalent in military veterans. However, few studies have investigated impulsivity as a risk factor for engaging in substance use behavior for individuals who are experiencing PTSD symptoms. The present study evaluated impulsivity as a moderator of the association between PTSD symptoms and alcohol/drug use. Male military veterans (N = 106) completed self-report measures of alcohol use behavior, drug use behavior, and impulsivity. Participants also completed a structured diagnostic interview to assess for PTSD. The findings indicated that impulsivity moderated the relation between total PTSD symptoms and alcohol use, B = 0.01, p = .035, along with associations between alcohol use and two of the symptom clusters: PTSD reexperiencing symptoms, B = 0.01, p = .016; and PTSD avoidance/numbing symptoms, B = 0.01, p = .029. Veterans with high levels of impulsivity were at significantly higher risk of engaging in alcohol use than veterans with low-to-average levels. Impulsivity did not potentiate the relation between PTSD hyperarousal symptoms and alcohol use nor did it moderate the association between any of the PTSD variables and drug use. Impulsivity appears to serve as a significant risk factor for alcohol use, but not drug use, for male veterans experiencing PTSD symptoms. Future studies are necessary to replicate and expand upon these findings, particularly to facilitate the development of integrated evidence-based treatments that target both alcohol use and impulsivity within the context of PTSD.

Intermittent Hypoxic Conditioning Alleviates Post-Traumatic Stress Disorder-Induced Damage and Dysfunction of Rat Visceral Organs and Brain.

Posttraumatic stress disorder (PTSD) causes mental and somatic diseases. Intermittent hypoxic conditioning (IHC) has cardio-, vaso-, and neuroprotective effects and alleviates experimental PTSD. IHC's ability to alleviate harmful PTSD effects on rat heart, liver, and brain was examined. PTSD was induced by 10-day exposure to cat urine scent (PTSD rats). Some rats were then adapted to 14-day IHC (PTSD+IHC rats), while PTSD and untreated control rats were cage rested. PTSD rats had a higher anxiety index (AI, X-maze test), than control or PTSD+IHC rats. This higher AI was associated with reduced glycogen content and histological signs of metabolic and hypoxic damage and of impaired contractility. The livers of PTSD rats had reduced glycogen content. Liver and blood alanine and aspartate aminotransferase activities of PTSD rats were significantly increased. PTSD rats had increased norepinephrine concentration and decreased monoamine oxidase A activity in cerebral cortex. The PTSD-induced elevation of carbonylated proteins and lipid peroxidation products in these organs reflects oxidative stress, a known cause of organ pathology. IHC alleviated PTSD-induced metabolic and structural injury and reduced oxidative stress. Therefore, IHC is a promising preventive treatment for PTSD-related morphological and functional damage to organs, due, in part, to IHC's reduction of oxidative stress.

Telomere shortening in blood leukocytes of patients with posttraumatic stress disorder.

Telomeres are protective fragments on chromosome ends involved in maintaining genome stability, preventing chromosomal fusions, regulation of cell division. It was shown that telomere attrition rate is accelerated in age-related diseases, as well as in response to physiological and psychosocial stress. The aim of this study was to evaluate relative leukocyte telomere length (LTL) in patients with post traumatic stress disorder (PTSD), as well as to investigate association of functional SNPs of telomerase TERC and TERT genes with LTL and PTSD. The relative LTL was measured by multiplex quantitative PCR method; genotyping of TERC rs12696304, TERT rs7726159 and rs2736100 was performed by PCR with sequence specific primers. Comparison of LTL in diseased and healthy subjects showed that PTSD patients had shorter average LTL than controls. Also, the frequency and the carriage rate of the TERT rs2736100*T allele was higher in PTSD patients compared to controls. Overall our results are in line with previous research in different populations. Furthermore, we have demonstrated that rs2736100 of TERT gene was significantly associated with PTSD and the minor allele of this polymorphism may be considered as a risk factor for PTSD in the Armenian population.

Activity of Paraoxonase/Arylesterase and Butyrylcholinesterase in Peripheral Blood of Gulf War Era Veterans With Neurologic Symptom Complexes or Post-Traumatic Stress Disorder.

OBJECTIVE: Two groups of Gulf War era veterans, one exhibiting blurred vision, balance problems/dizziness, tremors/shaking, and speech difficulty and a second group with post-traumatic stress disorder (PTSD), but not the neurologic syndrome, were assessed for organophosphate-detoxifying enzyme paraoxonase/arylesterase (PON1) and its Q/R isoforms, butyrylcholinesterase (BuChE) and its U/A isoforms and cytokines. METHODS: Defibrinated peripheral blood was evaluated for enzymes and cytokines. RESULTS: Trends toward elevation of Th2 cytokines interleukin-4 (IL-4) and IL-13 were observed in subjects with neurologic syndrome. Neither the activities nor isoforms of the enzyme, the neurologic symptoms, nor PTSD had any relationship to wartime deployment to the theater of combat. CONCLUSION: The negative outcomes described above suggest that exposure to organophosphates or other agents normally detoxified by PON1 and BuChE may not have contributed significantly to neurologic components of Gulf War Illness.

DICER1 and microRNA regulation in post-traumatic stress disorder with comorbid depression.

DICER1 is an enzyme that generates mature microRNAs (miRNAs), which regulate gene expression post-transcriptionally in brain and other tissues and is involved in synaptic maturation and plasticity. Here, through genome-wide differential gene expression survey of post-traumatic stress disorder (PTSD) with comorbid depression (PTSD&Dep), we find that blood DICER1 expression is significantly reduced in cases versus controls, and replicate this in two independent cohorts. Our follow-up studies find that lower blood DICER1 expression is significantly associated with increased amygdala activation to fearful stimuli, a neural correlate for PTSD. Additionally, a genetic variant in the 3' un-translated region of DICER1, rs10144436, is significantly associated with DICER1 expression and with PTSD&Dep, and the latter is replicated in an independent cohort. Furthermore, genome-wide differential expression survey of miRNAs in blood in PTSD&Dep reveals miRNAs to be significantly downregulated in cases versus controls. Together, our novel data suggest DICER1 plays a role in molecular mechanisms of PTSD&Dep through the DICER1 and the miRNA regulation pathway.

Elevated salivary alpha amylase in adolescent sexual abuse survivors with posttraumatic stress disorder symptoms.

OBJECTIVE: Little is known regarding neuroendocrine responses in adolescent girls with posttraumatic stress disorder (PTSD) who have experienced sexual abuse. Therefore, we collected saliva samples three times daily for 3 days to assess concentrations of salivary alpha amylase (sAA) - a surrogate marker for autonomic nervous system (ANS) activity and, in particular, sympathetic activity - in sexually abused adolescent girls. METHODS: Twenty-four girls (mean age: 15±1.4 years) who had experienced recent sexual abuse (i.e., sexual abuse occurred 1-6 months prior to study enrollment) and 12 healthy comparison subjects (mean age: 14.8±1.3 years) completed a structured interview and assessments to ascertain symptoms of posttraumatic stress, then collected saliva at home upon awakening, 30 minutes after waking, and at 5 p.m. on three consecutive school days. RESULTS: For sexually abused girls, total PTSD symptoms were associated with higher overall morning levels of sAA (r[20]=0.51, p=0.02), a finding driven by intrusive symptoms (r[20]=0.43, p<0.05) and hyperarousal symptoms (r[20]=0.58, p=0.01). There were no significant differences in diurnal sAA secretion between the sexually abused girls and healthy comparison adolescents. CONCLUSIONS: Overall morning concentrations of sAA in sexually abused girls are associated with overall PTSD severity as well as symptoms of hyperarousal and intrusive symptoms, possibly reflecting symptom-linked increases in ANS tone. These data raise the possibility that alterations in ANS activity are related to the pathophysiology of sexual abuse-related PTSD in adolescent girls, and may inform therapeutic interventions (e.g., antiadrenergic medications).

Histone deacetylases in memory and cognition.

Over the past 30 years, lysine acetylation of histone and nonhistone proteins has become established as a key modulator of gene expression regulating numerous aspects of cell biology. Neuronal growth and plasticity are no exception; roles for lysine acetylation and deacetylation in brain function and dysfunction continue to be uncovered. Transcriptional programs coupling synaptic activity to changes in gene expression are critical to the plasticity mechanisms underlying higher brain functions. These transcriptional programs can be modulated by changes in histone acetylation, and in many cases, transcription factors and histone-modifying enzymes are recruited together to plasticity-associated genes. Lysine acetylation, catalyzed by lysine acetyltransferases (KATs), generally promotes cognitive performance, whereas the opposing process, catalyzed by histone lysine deacetylases (HDACs), appears to negatively regulate cognition in multiple brain regions. Consistently, mutation or deregulation of different KATs or HDACs contributes to neurological dysfunction and neurodegeneration. HDAC inhibitors have shown promise as a treatment to combat the cognitive decline associated with aging and neurodegenerative disease, as well as to ameliorate the symptoms of depression and posttraumatic stress disorder, among others. In this review, we discuss the evidence for the roles of HDACs in cognitive function as well as in neurological disorders and disease. In particular, we focus on HDAC2, which plays a central role in coupling lysine acetylation to synaptic plasticity and mediates many of the effects of HDAC inhibition in cognition and disease.

Elevation of 11ß-hydroxysteroid dehydrogenase type 2 activity in Holocaust survivor offspring: evidence for an intergenerational effect of maternal trauma exposure.

BACKGROUND: Adult offspring of Holocaust survivors comprise an informative cohort in which to study intergenerational transmission of the effects of trauma exposure. Lower cortisol and enhanced glucocorticoid sensitivity have been previously demonstrated in Holocaust survivors with PTSD, and in offspring of Holocaust survivors in association with maternal PTSD. In other work, reduction in the activity of the enzyme 11ß-hydroxysteroid dehydrogenase type 2 (11ß-HSD-2), which inactivates cortisol, was identified in Holocaust survivors in comparison to age-matched, unexposed Jewish controls. Therefore, we investigated glucocorticoid metabolism in offspring of Holocaust survivors to evaluate if similar enzymatic decrements would be observed that might help to explain glucocorticoid alterations previously shown for Holocaust offspring. METHODS: Holocaust offspring (n=85) and comparison subjects (n=27) were evaluated with clinical diagnostic interview and self-rating scales, and asked to collect a 24-h urine sample from which concentrations of cortisol and glucocorticoid metabolites were assayed by GCMS. 11ß-HSD-2 activity was determined as the ratio of urinary cortisone to cortisol. RESULTS: Significantly reduced cortisol excretion was observed in Holocaust offspring compared to controls (p=.046), as had been shown for Holocaust survivors. However, 11ß-HSD-2 activity was elevated for offspring compared to controls (p=.008), particularly among those whose mothers had been children, rather than adolescents or adults, during World War II (p=.032). The effect of paternal Holocaust exposure could not be reliably investigated in the current sample. CONCLUSIONS: The inverse association of offspring 11ß-HSD-2 activity with maternal age at Holocaust exposure is consistent with the influence of glucocorticoid programming. Whereas a long standing reduction in 11ß-HSD-2 activity among survivors is readily interpreted in the context of Holocaust related deprivation, understanding the directional effect on offspring will require replication and further exploration.

The role of DNA methylation in stress-related psychiatric disorders.

Epigenetic modifications in response to traumatic experience and stress are emerging as important factors in the long-term biological trajectories leading to stress-related psychiatric disorders, reflecting both environmental influences as well as individual genetic predisposition. In particular, recent evidence on DNA methylation changes within distinct genes and pathways but also on a genome-wide level provides new insights into the pathophysiology of stress related psychiatric disorders. This review summarizes current findings and concepts on DNA methylation changes in stress-related disorders with a focus on major depressive disorder and posttraumatic stress disorder (PTSD). We highlight studies of DNA methylation in animals and humans pertinent to these disorders, both focusing on candidate loci as well as genome-wide studies. We describe molecular mechanisms of how exposure to stress can induce long lasting changes in DNA methylation and how these may relate to the pathophysiology of depression and PTSD. We discuss data suggesting that DNA methylation, even in peripheral tissues, appears to be an informative reflection of environmental exposures on the genome and may have potential as a biomarker for the early prevention of stress-related disorders.

[Effects of electroacupuncture on hippocampal nNOS expression in rats of post-traumatic stress disorder model].

OBJECTIVE: To explore the mechanism of electroacupuncture (EA) in the treatment of post-traumatic stress disorder (PTSD). METHODS: Thirty male Sprague-Dawley rats were randomly divided into a normal group, a model group and an electroacupuncture group. The single prolonged stress (SPS) method was used to set up the PTSD models in latter two groups. After SPS Stimulation, EA group was treated with 2Hz electroacupuncture at Baihui (GV 20) and Zusanli (ST 36) for 30 min, once a day for a week. Reverse transcriptase polymerase chain reaction (RT-PCR) and immuno-histochemistry were used to detect the mRNA and protein expression of nNOS in the hippocampus of rats in the each group. RESULTS: (1) The nNOS mRNA expression in hippocampus in model group was higher than that in normal group (P < 0.05). But the expression in EA group was lower significantly than that in model group (P < 0.05). (2) The nNOS protein expression in hippocampus CA1 and CA3 in model group was higher than that in normal group (P < 0.05). But after electroacupuncture treatment, its expression in EA group was lower significantly than that in model group (P < 0.05). The nNOS protein expression in hippocampal CA2 had no difference among all three groups. CONCLUSION: The elevated nNOS expression in hippocampus may be involved in the pathological process of PTSD. Electroacupuncture play a down-regulation effects in the hippocampal nNOS expression, which may be one mechanism of electroacupuncture for treatment of PTSD.

Gastrodin ameliorates anxiety-like behaviors and inhibits IL-1beta level and p38 MAPK phosphorylation of hippocampus in the rat model of posttraumatic stress disorder.

Gastrodin, a main constituent of a Chinese herbal medicine, has been shown to be effective in treating various mood disorders. The purpose of the present study was to determine whether gastrodin could ameliorate stress-associated behavior in a rat model of enhanced single prolonged stress (ESPS)-induced posttraumatic stress disorder (PTSD). Following ESPS, rats were administered orally with gastrodin (50, 100, or 200 mg/kg daily) or vehicle for 2 weeks. Animals were then tested in the open field and elevated plus-maze, and the levels of IL-6 and IL-1beta, the expression of iNOS, p38 and phospho-p38 (p-p38) in hippocampus were also tested. ESPS exposure resulted in pronounced anxiety-like behavior, elevated IL-6 and IL-1beta levels, and the higher expression of iNOS and p-p38 in hippocampus. However, repeated treatment with gastrodin, particularly at higher doses, reversed the aforementioned changes, including anxiety-like behavior, levels of IL-6 and IL-1beta, and the expression of iNOS and the p38 MAPK phosphorylation. These results indicate that gastrodin possesses anxiolytic effect and may be an effective herbal preparation for the treatment of PTSD.

Effect of calreticulin on Ca2+/CaM kinaseIIα and endoplasmic reticulum stress in hippocampal in a rat model of post-traumatic stress disorder.

The purpose of the present study was to examine the changes in the molecular chaperone calreticulin (CRT), calcium signaling pathway Ca(2+)-calmodulin (CaM)-CaM kinaseIIα (CaMKIIα), and the endoplasmic reticulum (ER) apoptotic modulator caspase-12 in hippocampal neurons of rats exposed to single-prolonged stress (SPS), a model of post-traumatic stress disorder (PTSD). Molecular markers and proteins were assessed using immunohistochemistry, western blot and reverse transcript-polymerase chain reaction in rats exposed to SPS at 1 day (1d), 4 and 7 days post-stress and time matched controls. We found that at 7 days, SPS rats had the highest CRT expression. The intracellular free Ca(2+) and the CaM expression reached peak at 1 day post-SPS whereas the CaMKIIα had the opposite trend. Caspase-12 was most active at 4 days and was found to decrease thereafter. Signs of apoptosis were identified using transmission electron microscopy in the rats exposed to SPS. The results indicate that signs of ER stress in the hippocampus of rats exposed to SPS trigger the molecular changes in the intracellular cytoplasm which in turn activate the apoptotic pathway through caspase-12. Therefore, we propose that the hippocampal apoptosis could be one of the pathological mechanisms related to the memory disorders in PTSD.

Enhanced Rho-kinase activity in patients with vasospastic angina after the Great East Japan Earthquake.

BACKGROUND: It remains unclear whether disease activity of vasospastic angina (VSA) is altered during a disaster. METHODS AND RESULTS: Before and after the Great East Japan Earthquake, we examined Rho-kinase activity in circulating neutrophils of 11 VSA patients and their mental stress with the post-traumatic stress disorder (PTSD) questionnaire. Rho-kinase activity was significantly increased at 6 months after the Earthquake, and was returned to baseline level at 12 months. Importantly, percent change in Rho-kinase activity was significantly correlated with the PTSD score. CONCLUSIONS: These results indicate that the Rho-kinase activity of VSA patients was transiently enhanced associated with disaster-related mental stress.

Altered salivary alpha-amylase awakening response in Bosnian War refugees with posttraumatic stress disorder.

In posttraumatic stress disorder (PTSD), chronic activation of the sympathetic nervous system (SNS) has been suggested. No study so far has investigated diurnal secretion patterns of salivary alpha-amylase (sAA) in PTSD, a promising candidate for non-invasive assessment of SNS activity. We compared sAA diurnal profiles between a group of Bosnian War refugees with PTSD and a healthy control group, and further analyzed for associations with psychiatric symptoms and glucocorticoid (GC) sensitivity of inflammatory regulation. PTSD patients showed a sAA awakening response profile that was opposite to those seen in healthy controls, i.e. an increase instead of a sharp decrease. Patterns of sAA secretion were further positively associated with psychiatric symptoms of PTSD. Finally, higher sAA awakening responses were associated with higher GC sensitivity of inflammatory cytokine production. These findings are in line with altered SNS function in PTSD, and lend further support for employing assessment of diurnal sAA profiles as non-invasive biomarkers in stress-related disease.

Single-prolonged stress induces increased phosphorylation of extracellular signal-regulated kinase in a rat model of post-traumatic stress disorder.

The extracellular signal-regulated kinase (ERK) signaling transduction pathway has been implicated in multiple physiological processes. It is not clear whether the ERK1/2 pathway participates in post-traumatic stress disorder (PTSD). The aim of this study was to provide novel insights into the mechanisms of how the amygdala participates in PTSD by investigating changes in the ERK1/2 pathway induced by single prolonged stress (SPS). The level of phosphorylated ERK1/2 (pERK1/2) protein was defined in a single-prolonged stress (SPS) animal model of post-traumatic stress disorder. A total of 100 male Wistar rats were randomly divided into a normal control group and SPS groups of 0, 30, 60 and 120 min. pERK1/2 distribution in the amygdala neurons was observed using immune electron microscopy. The expression of pERK1/2 was examined by immunohistochemistry and Western blotting. The pERK protein was located in some cell organelles, such as the mitochondria and neuraxon. Quantitatively, the expression of pERK protein level was significantly increased in the SPS rats. The results suggest that the ERK signal transduction pathway may play a crucial role in the pathology of PTSD.

Genotype-controlled analysis of serum dopamine ß-hydroxylase activity in civilian post-traumatic stress disorder.

BACKGROUND: Norepinephrine (NE) plays a central role in post-traumatic stress disorder (PTSD). Dopamine ß-hydroxylase (DßH) converts dopamine (DA) to NE and its activity varies widely across individuals. Mustapic et al. (2007) reported a PTSD-associated deficit in serum DßH activity in a genotype-controlled analysis of combat veterans. We tested whether such a deficit would occur in a sample of civilians. METHODS: The severity of current adult PTSD symptoms and current DSM-IV diagnosis of PTSD were determined by the PTSD Symptom Scale (PSS). Adulthood trauma exposure was assessed using the Traumatic Experience Inventory (TEI). Serum DßH activity (sDßH) was assayed by HPLC with electrochemical detection and genotypes were determined using the Taqman® platform. RESULTS: Two hundred and twenty seven African American (AA) subjects were enrolled in this study, with a mean age (±SD) of 42.9 (±12.9) years. We found a strong association between rs1611115 genotype and sDßH (p<0.0001). After controlling for adulthood trauma exposure, there were no significant differences of sDßH between subjects who met a PTSD diagnosis and those who did not (p>0.05) in any genotype group. No significant correlations were found between sDßH and PTSD severity, but sDßH significantly associated with the status of comorbid depression based on the cutoff of HAMD (p=0.014) in subjects with PTSD. CONCLUSIONS: We have replicated in this sample the prior finding that DBH rs1611115 genotype strongly associates with sDßH. No associations between sDßH and PTSD diagnosis or symptom severity were found in this civilian sample.