Beyond the Medial Regions of Prefrontal Cortex in the Regulation of Fear and Anxiety.

Fear and anxiety are adaptive responses but if left unregulated, or inappropriately regulated, they become biologically and socially maladaptive. Dysregulated emotions are manifest in a wide variety of psychiatric and neurological conditions but the external expression gives little indication of the underlying causes, which are inevitably multi-determined. To go beyond the overt phenotype and begin to understand the causal mechanisms leading to conditions characterized by anxiety and disorders of mood, it is necessary to identify the base psychological processes that have become dysregulated, and map them on to their associated neural substrates. So far, attention has been focused primarily on the medial regions of prefrontal cortex (PFC) and in particular their contribution to the expression and extinction of conditioned fear. However, functional neuroimaging studies have shown that the sphere of influence within the PFC is not restricted to its medial regions, but extends into dorsal, ventrolateral (vlPFC) and orbitofrontal (OFC) regions too; although the causal role of these other areas in the regulation of fear and anxiety remains to be determined and in the case of the OFC, existing findings are conflicting. Here, we review the evidence for the contribution of these other regions in negative emotion regulation in rodents and old world and new world monkeys. We consider a variety of different contexts, including conditioned and innate fear, learned and unlearned anxiety and cost-benefit decision-making, and a range of physiological and behavioral measures of emotion. It is proposed that both the OFC and vlPFC contribute to emotion regulation via their involvement, respectively, in the prediction of future outcomes and higher-order attentional control. The fractionation of these neurocognitive and neurobehavioral systems that regulate fear and anxiety opens up new opportunities for diagnostic stratification and personalized treatment strategies.This research was supported by a Medical Research Programme Grant (G0901884) from the Medical Research Council (MRC), UK to ACR and carried out within the Behavioral and Clinical Neurosciences Institute supported by a consortium award from the Wellcome Trust and the MRC.This is the final version of the article. It first appeared from Frontiers via http://dx.doi.org/10.3389/fnsys.2016.0001

Lesions of either anterior orbitofrontal cortex or ventrolateral prefrontal cortex in marmoset monkeys heighten innate fear and attenuate active coping behaviors to predator threat.

The ventral prefrontal cortex is an integral part of the neural circuitry that is dysregulated in mood and anxiety disorders. However, the contribution of its distinct sub-regions to the regulation of negative emotion are poorly understood. Recently we implicated both the ventrolateral prefrontal cortex (vlPFC) and anterior orbitofrontal cortex (antOFC) in the regulation of conditioned fear and anxiety responses to a social stimulus, i.e., human intruder, in the marmoset monkey. In the present study we extend our investigations to determine the role of these two regions in regulating innate responses and coping strategies to a predator stimulus, i.e., a model snake. Both the vlPFC and antOFC lesioned groups exhibited enhanced anxiety-related responses to the snake in comparison to controls. Both groups also showed a reduction in active coping behavior. These results indicate that the vlPFC and antOFC contribute independently to the regulation of both innate fear and, as previously reported, conditioned fear, and highlight the importance of these regions in producing stimulus-appropriate coping responses. The finding that dysregulation in two distinct prefrontal regions produces the apparently similar behavioral phenotype of heightened negative emotion provides insight into the varied etiology that may underlie this symptom across a wide variety of neuropsychiatric conditions with implications for personalized treatment strategies.This research was supported by a Medical Research Programme Grant (G0901884) from the Medical Research Council (MRC), UK to Angela C. Roberts. Yoshiro Shiba was supported by the Long Term Student Support Program provided by Osaka University and the Ministry of Education, Culture, Sports, Science and Technology of Japan and currently by the MRC Programme grant (G0901884). Andrea M. Santangelo, until October 2011, by a J. S. McDonnell Foundation grant (Principle Investigators; E. Phelps, T. W. Robbins, co-investigators; J. E. LeDoux, and Angela C. Roberts) and currently by the MRC Programme grant (G0901884). Work was carried out within the Behavioral and Clinical Neurosciences Institute supported by a consortium award from the Wellcome Trust and the MRC. We thank Dr. Carmen Agustín-Pavón for conducting the lesion surgeries, Dr. Katrin Braesicke for help with statistical analyses and Dr. Mercedes Arroyo for the preparation of histological material.This is the final published version of the paper, which first appeared in Frontiers Systems Neuroscience, 21 January 2015, doi: 10.3389/fnsys.2014.0025

A dimensional approach to modeling symptoms of neuropsychiatric disorders in the marmoset monkey.

Some patients suffering from the same neuropsychiatric disorder may have no overlapping symptoms whilst others may share symptoms common to other distinct disorders. Therefore, the Research Domain Criteria initiative recognises the need for better characterisation of the individual symptoms on which to focus symptom-based treatment strategies. Many of the disorders involve dysfunction within the prefrontal cortex (PFC) and so the marmoset, due to their highly developed PFC and small size, is an ideal species for studying the neurobiological basis of the behavioural dimensions that underlie these symptoms.Here we focus on a battery of tests that address dysfunction spanning the cognitive (cognitive inflexibility and working memory), negative valence (fear generalisation and negative bias) and positive valence (anhedonia) systems pertinent for understanding disorders such as ADHD, Schizophrenia, Anxiety, Depression and OCD. Parsing the separable prefrontal and striatal circuits and identifying the selective neurochemical modulation (serotonin vs dopamine) that underlie cognitive dysfunction have revealed counterparts in the clinical domain. Aspects of the negative valence system have been explored both at individual- (trait anxiety and genetic variation in serotonin transporter) and circuit-based levels enabling the understanding of generalisation processes, negative biases and differential responsiveness to SSRIs. Within the positive valence system, the combination of cardiovascular and behavioural measures provides a framework for understanding motivational, anticipatory and consummatory aspects of anhedonia and their neurobiological mechanisms. Together, the direct comparison of experimental findings in marmosets with clinical studies is proving an excellent translational model to address the behavioural dimensions and neurobiology of neuropsychiatric symptoms. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 328-353, 2017.This work was supported by long term funding from the MRC (ACR) and the Wellcome Trust (TWR) and was performed within the Behavioural and Clinical Neuroscience Institute, University of Cambridge, funded jointly by the Wellcome Trust and MRC. LO is supported by an MRC studentship.This is the author accepted manuscript. The final version is available from Wiley via http://dx.doi.org/10.1002/dneu.2244

Serotonergic, brain volume and attentional correlates of trait anxiety in primates.

Trait anxiety is a risk factor for the development and maintenance of affective disorders, and insights into the underlying brain mechanisms are vital for improving treatment and prevention strategies. Translational studies in non-human primates, where targeted neurochemical and genetic manipulations can be made, are critical in view of their close neuroanatomical similarity to humans in brain regions implicated in trait anxiety. Thus, we characterised the serotonergic and regional brain volume correlates of trait-like anxiety in the marmoset monkey. Low- and high-anxious animals were identified by behavioral responses to a human intruder (HI) that are known to be sensitive to anxiolytic drug treatment. Extracellular serotonin levels within the amygdala were measured with in vivo microdialysis, at baseline and in response to challenge with the selective serotonin reuptake inhibitor, citalopram. Regional brain volume was assessed by structural magnetic resonance imaging. Anxious individuals showed persistent, long-term fearful responses to both a HI and a model snake, alongside sustained attention (vigilance) to novel cues in a context associated with unpredictable threat. Neurally, high-anxious marmosets showed reduced amygdala serotonin levels, and smaller volumes in a closely connected prefrontal region, the dorsal anterior cingulate cortex. These findings highlight behavioral and neural similarities between trait-like anxiety in marmosets and humans, and set the stage for further investigation of the processes contributing to vulnerability and resilience to affective disorders.This research was supported by a Medical Research Programme Grant (G0901884) from the Medical Research Council UK (MRC) to Angela Roberts, and a PhD studentship from MRC and final-term funding from Trinity College, Cambridge, UK to Yevheniia Mikheenko.This is the author accepted manuscript. The final version is available from NPG at http://www.nature.com/npp/journal/v40/n6/full/npp2014324a.htm

Novel Primate Model of Serotonin Transporter Genetic Polymorphisms Associated with Gene Expression, Anxiety and Sensitivity to Antidepressants

This is the final version of the article. It first appeared from Nature Publishing Group via https://dx.doi.org/10.1038/npp.2016.41Genetic polymorphisms in the repeat upstream region of the serotonin transporter gene (SLC6A4) are associated with individual differences in stress reactivity, vulnerability to affective disorders and response to pharmacotherapy. However, the molecular, neurodevelopmental and psychopharmacological mechanisms underlying the link between SLC6A4 polymorphisms and the emotionally vulnerable phenotype are not fully understood. Thus, using the marmoset monkey Callithrix jacchus we characterize here a new neurobiological model to help to address these questions. We first sequenced the marmoset SLC6A4 promoter and identified a double nucleotide polymorphism (−2053AC/CT) and two single nucleotide polymorphisms (−2022C/T and −1592G/C) within the repeat upstream region. We showed their association with gene expression using in vivo quantitative PCR and with affective behavior using a primate test of anxiety (human intruder test). The low-expressing haplotype (AC/C/G) was linked with high anxiety whilst the high-expressing one (CT/T/C) was associated with an active coping strategy in response to threat. Pharmacological challenge with an acute dose of the selective serotonin reuptake inhibitor (SSRI), citalopram, revealed a genotype-dependent behavioral response. Whilst individuals homozygous for the high anxiety-related haplotype AC/C/G exhibited a dose-dependent, anxiogenic response, individuals homozygous for the low anxiety-related haplotype CT/T/C showed an opposing, dose-dependent anxiolytic effect. These findings provide a novel genetic and behavioral primate model to study the molecular, neurodevelopmental and psychopharmacological mechanisms that underlie genetic variation-associated complex behaviors, with specific implications for the understanding of normal and abnormal serotonin actions and the development of personalized pharmacological treatments for psychiatric disorders.Work was supported by an MRC Programme (ACR; G0901884) and performed within the Behavioural and Clinical Neuroscience Institute, University of Cambridge, funded jointly by the Wellcome Trust and MRC. AMS was supported by a McDonnell Foundation grant (PI’s: E. Phelps, T.W. Robbins; Co-Investigators: ACR and J. LeDoux; 22002015501) and currently supported by MRC; YS supported by the Long Term Student Support Program provided by Osaka University and the Ministry of Education, Culture, Sports, Science and Technology of Japan; HC supported by MRC Career Development Award and ACFS/MI supported by grants from the MRC and Wellcome Trust. GC supported by the Behavioural and Clinical Neuroscience Institute, Cambridge, United Kingdom. EHSS was self-funded

Characterising trait anxiety in the common marmoset (Callithrix jacchus): investigations into behavioural, psychophysiological and cognitive phenotypes

The major aim of my thesis project has been to develop a non-human primate model of trait anxiety, using a new world monkey, the common marmoset. The first step was to identify animals high or low in trait anxiety. Based on the findings that high trait-anxious individuals display over-generalization of fear responses, a pathogenic marker of elevated trait anxiety in humans, a new aversive discriminative conditioning paradigm was designed. Testing a normal cohort of marmosets revealed that 26% of the animals displayed both behavioural and physiological signs of fear generalization, i.e. failure to discriminate safety from danger cues (‘failed’ group). The remaining 74% showed successful discrimination (‘passed’ group). Additional regression analysis on several behavioural and physiological responses early in training revealed two potential biomarkers of high trait anxiety in marmosets: suppressed baseline blood pressure, indicative of contextual effects, and hyper cue-specific vigilance. These measures predicted the animal’s likelihood of passing or failing the discrimination. The finding that the ‘failed’ group showed intact discriminative performance in the appetitive domain rules out an interpretation of the results in terms of a general impairment in learning, per se. To further determine whether these hypothetically high trait-anxious animals would display enhanced anxiety-related responses in more classical primate models of anxiety, human intruder and rubber snake tests were performed on a large sample of marmosets. Principal component analysis on multiple behavioural measures revealed two components underlying performance: ‘emotionality’ and ‘coping strategy’. Although no difference was found in the human intruder test, the ’failed’ group displayed significantly elevated levels of ‘emotionality’ in comparison to the ‘passed’ group in the rubber snake test. Moreover, the two biomarkers of fear over-generalisation also reliably predicted the ‘emotionality’ scores. Finally, having developed a marmoset model of trait anxiety, investigations into the neural underpinnings, especially prefrontal involvement in trait anxiety mechanisms, were carried out by testing the animals on two cognitive flexibility tests: an orbitofrontal cortex (OFC)-dependent incongruent object discrimination test and a lateral prefrontal cortex (lPFC)-dependent detour reaching rule transfer test. Whilst group differences did not reach significance, the two biomarkers of fear over-generalisation, the suppressed baseline blood pressure and hyper cue-specific vigilance, were inversely and differentially correlated with perseverative performance on the two tests, the lPFC- and OFC-dependent tests, respectively. This not only indicates that high trait anxiety can lead to improvements in certain aspects of prefrontal cognitive function but also suggests that changes in the activity of at least two distinct prefronto-subcortical neural circuits, a cue-sensitive amygdala-OFC and a context-sensitive hippocampus-lPFC circuit, may contribute to trait anxiety

Beyond the Medial Regions of Prefrontal Cortex in the Regulation of Fear and Anxiety

This is the final version of the article. It first appeared from Frontiers via http://dx.doi.org/10.3389/fnsys.2016.00012Fear and anxiety are adaptive responses but if left unregulated, or inappropriately regulated, they become biologically and socially maladaptive. Dysregulated emotions are manifest in a wide variety of psychiatric and neurological conditions but the external expression gives little indication of the underlying causes, which are inevitably multi-determined. To go beyond the overt phenotype and begin to understand the causal mechanisms leading to conditions characterized by anxiety and disorders of mood, it is necessary to identify the base psychological processes that have become dysregulated, and map them on to their associated neural substrates. So far, attention has been focused primarily on the medial regions of prefrontal cortex (PFC) and in particular their contribution to the expression and extinction of conditioned fear. However, functional neuroimaging studies have shown that the sphere of influence within the PFC is not restricted to its medial regions, but extends into dorsal, ventrolateral (vlPFC) and orbitofrontal (OFC) regions too; although the causal role of these other areas in the regulation of fear and anxiety remains to be determined and in the case of the OFC, existing findings are conflicting. Here, we review the evidence for the contribution of these other regions in negative emotion regulation in rodents and old world and new world monkeys. We consider a variety of different contexts, including conditioned and innate fear, learned and unlearned anxiety and cost-benefit decision-making, and a range of physiological and behavioral measures of emotion. It is proposed that both the OFC and vlPFC contribute to emotion regulation via their involvement, respectively, in the prediction of future outcomes and higher-order attentional control. The fractionation of these neurocognitive and neurobehavioral systems that regulate fear and anxiety opens up new opportunities for diagnostic stratification and personalized treatment strategies.This research was supported by a Medical Research Programme Grant (G0901884) from the Medical Research Council (MRC), UK to ACR and carried out within the Behavioral and Clinical Neurosciences Institute supported by a consortium award from the Wellcome Trust and the MRC