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Memory reconsolidation prevents extinction of learned immunosuppression
Abstracts / Brain, Behavior, and Immunity 49 (2015) e1–e50
War field is a complicated stress environment, in which psychological stress and fatigue cumulates. The purpose of this study is to set up war field exposure stress (WS) model and to investigate restitution effects of biologically-active substances on psychoneuroimmunological regulation disturbed by WS. War-field-like environment was constructed with rotational cage to interfere sleep, metal-meshed bottom to generate electrical shock, and speaker to provide sound stress. Each experimental animal was exposed to 24 hours of stress 3 times in 2 weeks. 2 times of electric shocks and sound shocks were randomly involved in a 24-hour stress exposure. Exposure to WS increased significantly motor activity and anxiety-like behavior in mice (p < :03, p < :02, respectively). Serum corticosterone and proinflammatory cytokines, IFNc and TNFa, in supernatant of T cell culture extracted from mice exposed to WS were significantly higher than that of control mice (p < :01, p < :001, p < :01, respectively). Anxiolytic drug, adaptol and ginsenoside Rg1, had no effect on hyper-motor activity and anxiety-like behavior of mice exposed to WS. However, adaptol and ginsenoside Rg1 down-regulated serum corticosterone level (p < :04) and IFNc and TNFa in supernatant of T cell culture extracted from mice exposed to WS (p < :01, p < :02, respectively). This study suggests that neurobehavioral and immunological adaptation to stress may have different pathway, and adaptol and ginsenoside Rg1 may have no effects on anxiety-like behavior caused by stress. http://dx.doi.org/10.1016/j.bbi.2015.06.044
Abstract # 1532 Selective manipulation of spinal microglia by chemogenetics: Implications for allodynia and inflammatory signaling P.M. Grace a, X. Wang a, D.J. Urban b, M.V. Baratta a, E.L. Galer a, K.A. Strand a, Y. Zhang a, H. Yin a, B.L. Roth b, S.F. Maier a, L.R. Watkins a a
University of Colorado Boulder, Psychology and Neuroscience, UCB 345, Boulder, CO 80309, United States b University of North Carolina, Chapel Hill, United States The absence of selective pharmacological tools is a major barrier to the in vivo study of microglia. To address this issue, we developed a Gq- and Gi-coupled Designer Receptor Exclusively Activated by a Designer Drug (DREADD) to enable selective stimulation or inhibition of microglia, respectively. DREADDs under a CD68 (microglia/macrophage) promoter were intrathecally transfected via an AAV9 vector. Naïve rats intrathecally transfected with Gq (stimulatory) DREADDs exhibited significant allodynia following intrathecal administration of the DREADD-selective ligand clozapine-N-oxide (CNO), which was abolished by intrathecal IL-1ra. Chronic constriction injury-induced allodynia was attenuated by intrathecal CNO in rats intrathecally transfected with Gi (inhibitory) DREADDs. To explore mechanisms, BV2 cells were stably transfected with Gq or Gi DREADDs in vitro. CNO treatment induced pro-inflammatory mediator production per se from cells expressing Gq-DREADDs, and inhibited lipopolysaccharide-induced pro-inflammatory mediator production from cells expressing Gi-DREADDs. These studies are the first to manipulate microglia using DREADDs, which allow the role of glia in pain to be conclusively demonstrated, unconfounded by neuronal off-target effects that exist for all other glial inhibitors. Hence, these studies are the first to conclusively demonstrate that in vivo stimulation of resident spinal microglia in intact spinal cord is (a) sufficient for allodynia, and (b) necessary for allodynia induced
e7
by peripheral nerve injury. DREADDs are a unique tool to selectively explore the physiological and pathological role of microglia in vivo. http://dx.doi.org/10.1016/j.bbi.2015.06.045
Abstract # 1533 Exercise alters the gut microbiome and microbial metabolites: Implications for colorectal cancer and inflammatory bowel disease J.A. Woods a,b,c, J.M. Allen a,b, M.E. Berg Miller d,e, B.A. White c,d,e, H. Gaskins c,d,e, V. Nehra f a Department of Kinesiology and Community Health, 906 S. Goodwin Ave., University of Illinois at Urbana, Urbana, IL 61801, United States b Integrative Immunology and Behavior Program, United States c Division of Nutritional Sciences, United States d Department of Animal Sciences, United States e Institute for Genomic Biology, United States f Department of Gastroenterology, Mayo Clinic, Rochester, MN, United States
Introduction: Exercise training reduces colon cancer and inflammatory bowel diseases (IBD), but the mechanisms remain unknown. We hypothesized that acute and chronic exercise would alter the gut microbiota and its associated metabolome in a beneficial way. Methods: Mice exercised for 6 weeks with access to voluntary wheels (VWR; n = 10), or were forcibly run on a treadmill running (FTR; n = 10). A control group remained sedentary (SED; n = 9). The conserved bacterial 16s rRNA gene was isolated from the feces and cecum and sequenced using high-throughput Illumina Miseq. In another experiment, mice were randomized to sedentary (SED, n = 10), acute moderate (MOD, n = 10) or high intensity running (HI, n = 10). Mice were euthanized 45 min post and cecal contents were analyzed for short chain fatty acids (SCFA) by GC-MS. Results: Exercise training differentially altered the community structure of the microbiome at both intestinal sites (p < 0:05). Further evaluation revealed that exercise altered many bacterial taxa in both the feces and the cecum. Notably, voluntary exercise reduced ceca and fecal concentrations of Turicibacter spp., a genus of bacteria implicated in ulcerative colitis in humans and mice. Acute, HI (but not MOD) exercise increased the levels of potentially beneficial acetate, propionate, and butyrate, compared to SED (p < 0:05). Conclusion: Acute and chronic exercise invokes changes in the microbiome and metabolome that may be beneficial to the prevention or treatment of IBD and colon cancer. http://dx.doi.org/10.1016/j.bbi.2015.06.046
Abstract # 1534 Memory reconsolidation prevents extinction of learned immunosuppression M. Schedlowski, M. Hadamitzky, K. Boesche, L. Lueckemann, H. Engler Institute of Medical Psychology & Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, Essen 45122, Germany When memories are recalled, they enter a transient labile phase in which they can be impaired or enhanced followed by a new
e8
Abstracts / Brain, Behavior, and Immunity 49 (2015) e1–e50
stabilization process termed reconsolidation. It is unknown, however, whether reconsolidation is restricted to neurocognitive processes such as fear memories or can be extended to peripheral physiological functions as well. Bidirectional interaction between the brain and peripheral immune system, together with the fact that immune functions can be modified by associative learning processes provide a unique opportunity to test this assumption. Employing an established paradigm of behaviorally-conditioned taste aversion in rats, we here demonstrate reconsolidation in ‘‘learned immunosuppression’’. The administration of sub-therapeutic doses of the calcineurin inhibitor and immunosuppressant cyclosporin A (CsA) together with the conditioned stimulus (CS/saccharine) during evocation blocked the extinction of conditioned taste aversion and learned suppression of T cell cytokine (interleukin-2; interferon-gamma) production. In contrast, reconsolidation of learned immunosuppression could not be induced by administering sub-therapeutic CsA as a reminder cue outside the reconsolidation window, 8 hours after CS exposure. This reconsolidated immunosuppression is of clinical relevance since it significantly prolonged the survival time of heterotopically-transplanted heart allografts in rats. Collectively, these findings demonstrate that memories can reconsolidate on peripheral physiological systems such as immune functioning; they might pave the way for the systematic integration of conditioning paradigms as supportive therapy in pharmacological regimens. http://dx.doi.org/10.1016/j.bbi.2015.06.047
Abstract # 1535 RNA microarray analysis in dried blood spots A. Danese Institute of Psychiatry, Psychology and Neuroscience, King’s College London, UK, 16 DeCrespigny Park, London SE5 8AF, United Kingdom Objectives: Despite the promises and ever-decreasing costs, the genomic revolution has so far had only a marginal role in epidemiological research. This is at least partly due to lack of minimally-invasive methods of specimen collection that can be used for genomic analysis. We aimed to address this gap by testing whether a minimally-invasive method of collection through dried blood spots provides a good proxy for microarray (RNA expression) analysis in whole blood. Methods: We collected venipuncture and dried blood spot samples from 19 individuals and performed whole genome expression analysis on extracted RNA using the Illumina platform. Results: We found a correlation of rho = .76 and an agreement of kappa = .45 for detected genes in the two datasets (validity). We found a correlation of rho = .96 and an agreement of kappa = .72 for detected genes in the independent replicates from dried blood spots (reliability). Conclusions: Genome-wise RNA expression in dried blood spots shows a moderate-to-high correlation and fair-to-good agreement with RNA expression in whole blood. If these findings are replicated in larger samples, microarray analysis in dried blood spots could be used as a minimally-invasive method to assess genomic underpinning of illness in population studies. http://dx.doi.org/10.1016/j.bbi.2015.06.048
Abstract # 1536 Effects of vagus nerve stimulation on neurophysiological parameters and the cellular immune response in the rat brain during endotoxemia H. Schweighöfer a, C. Rummel b, J. Roth b, B. Rosengarten a a
Department of Neurology, Justus-Liebig University Giessen, Giessen 35392, Germany b Institute of Veterinary-Physiology and Biochemistry, Justus-Liebig University Giessen, Germany Previous experiments indicated that stimulation of the vagus nerve has modulating, anti-inflammatory effects on the cellular immune response in the blood and the spleen, stabilising brain function. However, in these studies, underlying mechanisms remained largely unknown. Here, we aimed to investigate the potential effects of vagus nerve-stimulation for immune-to-brain communication focused on neurophysiological readouts and leukocyte migration to the brain during severe septic-like endotoxemia. Systemic inflammation was induced by intravenous administration of lipopolysaccaride (LPS; 5 mg/kg). Animals received either no manipulation of the vagus nerve, vagotomy or vagotomy plus vagus nerve stimulation of the distal trunk. Somatosensory evoked potentials and evoked flow velocity response were measured for 4.5 h as indicators of brain function and neurovascular coupling, respectively. LPS induced a decline of both, which was recovered by vagus nerve stimulation. As for peripheral organs, LPS-stimulated neutrophil counts increased in the brain and colocalized with intercellular adhesion molecule (ICAM)-1. Interestingly, vagal stimulation reduced colocalisation between neutrophil granulocytes and ICAM-1 and decreased nuclear translocation of the brain cell activation marker nuclear factor interleukin (NF-IL)6. Furthermore it reduced the gene expression of inflammatory markers and extravasation signals (IL-6, CXCL-1, ICAM-1) in the hypothalamus. Overall, our findings suggest beneficial, anti-inflammatory effects of vagus nerve stimulation on neurophysiological parameters that potentially are due to reduced interaction of neutrophil granulocytes with brain endothelial cells and an attenuated inflammatory response of the brain. http://dx.doi.org/10.1016/j.bbi.2015.06.049
Abstract # 1537 Generalist association psychopathology A. Danese
between
inflammation
and
Institute of Psychiatry, Psychology and Neuroscience, King’s College London, 16 DeCrespigny Park, London SE5 8AF, Great Britain and Northern Ireland, UK Objectives: Several different psychiatric diagnoses have been linked to high levels of circulating inflammation biomarkers. However, it is unclear how patterns of concurrent and sequential comorbidity impact on these findings. We tested whether the association between different psychiatric diagnoses and inflammation is accounted for by diagnosis-specific effects or by a higher-order, non-specific liability to general psychopathology that captures patterns of concurrent and sequential comorbidity. Methods: We tested this hypothesis in data from the Dunedin Multidisciplinary Health and Development Study, a
War field is a complicated stress environment, in which psychological stress and fatigue cumulates. The purpose of this study is to set up war field exposure stress (WS) model and to investigate restitution effects of biologically-active substances on psychoneuroimmunological regulation disturbed by WS. War-field-like environment was constructed with rotational cage to interfere sleep, metal-meshed bottom to generate electrical shock, and speaker to provide sound stress. Each experimental animal was exposed to 24 hours of stress 3 times in 2 weeks. 2 times of electric shocks and sound shocks were randomly involved in a 24-hour stress exposure. Exposure to WS increased significantly motor activity and anxiety-like behavior in mice (p < :03, p < :02, respectively). Serum corticosterone and proinflammatory cytokines, IFNc and TNFa, in supernatant of T cell culture extracted from mice exposed to WS were significantly higher than that of control mice (p < :01, p < :001, p < :01, respectively). Anxiolytic drug, adaptol and ginsenoside Rg1, had no effect on hyper-motor activity and anxiety-like behavior of mice exposed to WS. However, adaptol and ginsenoside Rg1 down-regulated serum corticosterone level (p < :04) and IFNc and TNFa in supernatant of T cell culture extracted from mice exposed to WS (p < :01, p < :02, respectively). This study suggests that neurobehavioral and immunological adaptation to stress may have different pathway, and adaptol and ginsenoside Rg1 may have no effects on anxiety-like behavior caused by stress. http://dx.doi.org/10.1016/j.bbi.2015.06.044
Abstract # 1532 Selective manipulation of spinal microglia by chemogenetics: Implications for allodynia and inflammatory signaling P.M. Grace a, X. Wang a, D.J. Urban b, M.V. Baratta a, E.L. Galer a, K.A. Strand a, Y. Zhang a, H. Yin a, B.L. Roth b, S.F. Maier a, L.R. Watkins a a
University of Colorado Boulder, Psychology and Neuroscience, UCB 345, Boulder, CO 80309, United States b University of North Carolina, Chapel Hill, United States The absence of selective pharmacological tools is a major barrier to the in vivo study of microglia. To address this issue, we developed a Gq- and Gi-coupled Designer Receptor Exclusively Activated by a Designer Drug (DREADD) to enable selective stimulation or inhibition of microglia, respectively. DREADDs under a CD68 (microglia/macrophage) promoter were intrathecally transfected via an AAV9 vector. Naïve rats intrathecally transfected with Gq (stimulatory) DREADDs exhibited significant allodynia following intrathecal administration of the DREADD-selective ligand clozapine-N-oxide (CNO), which was abolished by intrathecal IL-1ra. Chronic constriction injury-induced allodynia was attenuated by intrathecal CNO in rats intrathecally transfected with Gi (inhibitory) DREADDs. To explore mechanisms, BV2 cells were stably transfected with Gq or Gi DREADDs in vitro. CNO treatment induced pro-inflammatory mediator production per se from cells expressing Gq-DREADDs, and inhibited lipopolysaccharide-induced pro-inflammatory mediator production from cells expressing Gi-DREADDs. These studies are the first to manipulate microglia using DREADDs, which allow the role of glia in pain to be conclusively demonstrated, unconfounded by neuronal off-target effects that exist for all other glial inhibitors. Hence, these studies are the first to conclusively demonstrate that in vivo stimulation of resident spinal microglia in intact spinal cord is (a) sufficient for allodynia, and (b) necessary for allodynia induced
e7
by peripheral nerve injury. DREADDs are a unique tool to selectively explore the physiological and pathological role of microglia in vivo. http://dx.doi.org/10.1016/j.bbi.2015.06.045
Abstract # 1533 Exercise alters the gut microbiome and microbial metabolites: Implications for colorectal cancer and inflammatory bowel disease J.A. Woods a,b,c, J.M. Allen a,b, M.E. Berg Miller d,e, B.A. White c,d,e, H. Gaskins c,d,e, V. Nehra f a Department of Kinesiology and Community Health, 906 S. Goodwin Ave., University of Illinois at Urbana, Urbana, IL 61801, United States b Integrative Immunology and Behavior Program, United States c Division of Nutritional Sciences, United States d Department of Animal Sciences, United States e Institute for Genomic Biology, United States f Department of Gastroenterology, Mayo Clinic, Rochester, MN, United States
Introduction: Exercise training reduces colon cancer and inflammatory bowel diseases (IBD), but the mechanisms remain unknown. We hypothesized that acute and chronic exercise would alter the gut microbiota and its associated metabolome in a beneficial way. Methods: Mice exercised for 6 weeks with access to voluntary wheels (VWR; n = 10), or were forcibly run on a treadmill running (FTR; n = 10). A control group remained sedentary (SED; n = 9). The conserved bacterial 16s rRNA gene was isolated from the feces and cecum and sequenced using high-throughput Illumina Miseq. In another experiment, mice were randomized to sedentary (SED, n = 10), acute moderate (MOD, n = 10) or high intensity running (HI, n = 10). Mice were euthanized 45 min post and cecal contents were analyzed for short chain fatty acids (SCFA) by GC-MS. Results: Exercise training differentially altered the community structure of the microbiome at both intestinal sites (p < 0:05). Further evaluation revealed that exercise altered many bacterial taxa in both the feces and the cecum. Notably, voluntary exercise reduced ceca and fecal concentrations of Turicibacter spp., a genus of bacteria implicated in ulcerative colitis in humans and mice. Acute, HI (but not MOD) exercise increased the levels of potentially beneficial acetate, propionate, and butyrate, compared to SED (p < 0:05). Conclusion: Acute and chronic exercise invokes changes in the microbiome and metabolome that may be beneficial to the prevention or treatment of IBD and colon cancer. http://dx.doi.org/10.1016/j.bbi.2015.06.046
Abstract # 1534 Memory reconsolidation prevents extinction of learned immunosuppression M. Schedlowski, M. Hadamitzky, K. Boesche, L. Lueckemann, H. Engler Institute of Medical Psychology & Behavioral Immunobiology, University Hospital Essen, University of Duisburg-Essen, Essen 45122, Germany When memories are recalled, they enter a transient labile phase in which they can be impaired or enhanced followed by a new
e8
Abstracts / Brain, Behavior, and Immunity 49 (2015) e1–e50
stabilization process termed reconsolidation. It is unknown, however, whether reconsolidation is restricted to neurocognitive processes such as fear memories or can be extended to peripheral physiological functions as well. Bidirectional interaction between the brain and peripheral immune system, together with the fact that immune functions can be modified by associative learning processes provide a unique opportunity to test this assumption. Employing an established paradigm of behaviorally-conditioned taste aversion in rats, we here demonstrate reconsolidation in ‘‘learned immunosuppression’’. The administration of sub-therapeutic doses of the calcineurin inhibitor and immunosuppressant cyclosporin A (CsA) together with the conditioned stimulus (CS/saccharine) during evocation blocked the extinction of conditioned taste aversion and learned suppression of T cell cytokine (interleukin-2; interferon-gamma) production. In contrast, reconsolidation of learned immunosuppression could not be induced by administering sub-therapeutic CsA as a reminder cue outside the reconsolidation window, 8 hours after CS exposure. This reconsolidated immunosuppression is of clinical relevance since it significantly prolonged the survival time of heterotopically-transplanted heart allografts in rats. Collectively, these findings demonstrate that memories can reconsolidate on peripheral physiological systems such as immune functioning; they might pave the way for the systematic integration of conditioning paradigms as supportive therapy in pharmacological regimens. http://dx.doi.org/10.1016/j.bbi.2015.06.047
Abstract # 1535 RNA microarray analysis in dried blood spots A. Danese Institute of Psychiatry, Psychology and Neuroscience, King’s College London, UK, 16 DeCrespigny Park, London SE5 8AF, United Kingdom Objectives: Despite the promises and ever-decreasing costs, the genomic revolution has so far had only a marginal role in epidemiological research. This is at least partly due to lack of minimally-invasive methods of specimen collection that can be used for genomic analysis. We aimed to address this gap by testing whether a minimally-invasive method of collection through dried blood spots provides a good proxy for microarray (RNA expression) analysis in whole blood. Methods: We collected venipuncture and dried blood spot samples from 19 individuals and performed whole genome expression analysis on extracted RNA using the Illumina platform. Results: We found a correlation of rho = .76 and an agreement of kappa = .45 for detected genes in the two datasets (validity). We found a correlation of rho = .96 and an agreement of kappa = .72 for detected genes in the independent replicates from dried blood spots (reliability). Conclusions: Genome-wise RNA expression in dried blood spots shows a moderate-to-high correlation and fair-to-good agreement with RNA expression in whole blood. If these findings are replicated in larger samples, microarray analysis in dried blood spots could be used as a minimally-invasive method to assess genomic underpinning of illness in population studies. http://dx.doi.org/10.1016/j.bbi.2015.06.048
Abstract # 1536 Effects of vagus nerve stimulation on neurophysiological parameters and the cellular immune response in the rat brain during endotoxemia H. Schweighöfer a, C. Rummel b, J. Roth b, B. Rosengarten a a
Department of Neurology, Justus-Liebig University Giessen, Giessen 35392, Germany b Institute of Veterinary-Physiology and Biochemistry, Justus-Liebig University Giessen, Germany Previous experiments indicated that stimulation of the vagus nerve has modulating, anti-inflammatory effects on the cellular immune response in the blood and the spleen, stabilising brain function. However, in these studies, underlying mechanisms remained largely unknown. Here, we aimed to investigate the potential effects of vagus nerve-stimulation for immune-to-brain communication focused on neurophysiological readouts and leukocyte migration to the brain during severe septic-like endotoxemia. Systemic inflammation was induced by intravenous administration of lipopolysaccaride (LPS; 5 mg/kg). Animals received either no manipulation of the vagus nerve, vagotomy or vagotomy plus vagus nerve stimulation of the distal trunk. Somatosensory evoked potentials and evoked flow velocity response were measured for 4.5 h as indicators of brain function and neurovascular coupling, respectively. LPS induced a decline of both, which was recovered by vagus nerve stimulation. As for peripheral organs, LPS-stimulated neutrophil counts increased in the brain and colocalized with intercellular adhesion molecule (ICAM)-1. Interestingly, vagal stimulation reduced colocalisation between neutrophil granulocytes and ICAM-1 and decreased nuclear translocation of the brain cell activation marker nuclear factor interleukin (NF-IL)6. Furthermore it reduced the gene expression of inflammatory markers and extravasation signals (IL-6, CXCL-1, ICAM-1) in the hypothalamus. Overall, our findings suggest beneficial, anti-inflammatory effects of vagus nerve stimulation on neurophysiological parameters that potentially are due to reduced interaction of neutrophil granulocytes with brain endothelial cells and an attenuated inflammatory response of the brain. http://dx.doi.org/10.1016/j.bbi.2015.06.049
Abstract # 1537 Generalist association psychopathology A. Danese
between
inflammation
and
Institute of Psychiatry, Psychology and Neuroscience, King’s College London, 16 DeCrespigny Park, London SE5 8AF, Great Britain and Northern Ireland, UK Objectives: Several different psychiatric diagnoses have been linked to high levels of circulating inflammation biomarkers. However, it is unclear how patterns of concurrent and sequential comorbidity impact on these findings. We tested whether the association between different psychiatric diagnoses and inflammation is accounted for by diagnosis-specific effects or by a higher-order, non-specific liability to general psychopathology that captures patterns of concurrent and sequential comorbidity. Methods: We tested this hypothesis in data from the Dunedin Multidisciplinary Health and Development Study, a