Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - URBACH-ALZ (Hyper-emotionality after neurodegenerative loss of inhibition of the amygdala)

Teaser

\"The human subjects in South-Africa: a globally unique population.The group of human subjects that participated to this study suffer from Urbach Wiethe disease (UWD), an extremely rare genetic condition. In the Northern Cape of South Africa, however, it has spread as a result...

Summary

\"The human subjects in South-Africa: a globally unique population.
The group of human subjects that participated to this study suffer from Urbach Wiethe disease (UWD), an extremely rare genetic condition. In the Northern Cape of South Africa, however, it has spread as a result of a founder effect after introduction of a genetic mutation by European settlers 400 years ago. These UWD patients exhibit selective damage of a restricted part of brain called Basolateral amygdala (BLA), located in the temporal lobe. The adjacent brain region, the Central amygdala (CeA), which triggers different expressions of fear (freezing, potentiated startle, physiological changes), is intact. Thus, this is a unique occasion to assess a new function of the human BLA. Contrary to the previous UWD case, we found that these UWD subjects did not exhibit decreased fear, but instead increased fear responses, particularly when facing an escapable threat. This suggested an inhibitory control of the BLA over passive fear responses that are triggered by the CeA, and that becomes active when preparing to escape from threat.

A new cross-species mechanism for actively controlling fear.
To expose this precise mechanism, we 1) developed in parallel a rat model in which we inhibited the BLA with a recent technology, called chemogenetic, and 2) designed for both human subjects and rats an equivalent \"\"escape from threat\"\" task. We then assessed central amygdala output to the brainstem by measuring in rats freezing, a common animal reaction facing feargul stimuli, and both in rats and humans potentiated startle responses, a reflex conserved between humans and rodents. These measures are considered most direct readouts of passive responses to threats. In both species loss of BLA function increased these responses, and was accompanied, in rats, by a decrease in active escape from threat. In the UWD group this increased central amygdala output could also be measured by fMRI of the CeA and pons.
To assess the pathway through which the BLA controls output from the central amygdala to the brainstem we studied a local inhibitory pathway within the CeA that we had previously revealed to be oxytocin-sensitive. We found that its activation by oxytocin could fully recover the changes in freezing, startle and escape response caused by BLA inhibition. This thereby provides a new interpretation of our previous data that had shown decreased freezing by oxytocin without affecting physiological expression of fear. Thus, oxytocin appears to prepare the rat for escape, by actively facing the threat. Taken together, we identified a novel mechanism through which the BLA inhibits passive fear responses when the individual is offered the possibility to escape: It involves BLA activation of inhibitory neurons in the CeA that are sensitive to oxytocin.

Impact of the study.
1. Animal-human translation researchers
Our data reveal how the BLA, via the CeA, adaptively regulates escape behavior from imminent threat and that this mechanism is evolutionary conserved across rodents and humans. Putting an animal model side-by-side these human findings represents a rather unique combination of two fields, which we hope may push the boundaries for innovative translational research.
2. Human emotion and clinical researchers: The BLA in humans compared to rodents has undergone an expansion in size relative to other amygdala subregions that is huge and it may have gained additional functions to similar extent. We feel, in fact, we are only at the beginning of starting to explore the rich repertoire of changes in emotional and motivational behavior that is present in these BLA-damaged human subjects. The brain mechanism we explored, and specifically the pharmacological activation method employed, opens up new opportunities for the study and potential treatment of fear and anxiety disorders.
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Work performed

\"This project involved both human and animal Neuroscience, thus I coordinated the studies involving the experimental part in the animal model and the part regarding experiments in humans subjects in collaboration with Prof. Armin von Gunten (at SUPAA, CHUV) and Prof. Jack van Honk (at UCT, Cape Town) to compare results that were included in a joint publication in the scientific journal \"\"Cell\"\".

I have conducted the experiments on rodents to create an animal model with inactivated BLA to make a parallel with BLA-damaged patients. During this period, I have also worked on the development of a rodent version of the human “Threat and Escape Task” (TET). Then I tested the effects of BLA inactivation on the behavioral responses to imminent threat. To do this I have injected rats with a viral chemogenetic system (DREADDs) able to induce the neuronal silencing of the BçA. Finally I have tested the role of oxytocin in rats with and without BLA inactivation. For this purpose I used a combined approach involving pharmacological manipulation and electrophysiological measurements in the animal model.

In summary, in this project we compared humans with natural-selective bilateral BLA lesions to rats with a chemogenetically silenced BLA. We found, across species, an essential role for the BLA in the selection of active escape over passive freezing reaction during exposure to imminent yet escapable threat. In response to imminent threats, BLA-damaged humans showed increased startle potentiation and BLA-silenced rats demonstrated increased startle potentiation, freezing, and reduced escape behavior as compared to controls. Neuroimaging in humans suggested that the BLA reduces passive defensive responses by inhibiting the brainstem via the CeA. Indeed, imminent threat conditioning potentiated BLA projections onto an inhibitory CeA pathway, and pharmacological activation of this pathway rescued deficient imminent threats responses in BLA-silenced rats. In conclusion, our data reveal how the BLA, via the CeA, adaptively regulates escape behavior from imminent threat and that this mechanism is evolutionary conserved across rodents and humans.

The dissemination of the scientific results obtained during this project occurred both at local and international level. At a local level we presented the results at the Swiss Society for Neuroscience Annual meeting. Moreover, we showed our results at the most important meetings in the field of Neuroscience: the Annual Society for Neuroscience Meeting in 2017 and 2018 and the FENS Forum of Neuroscience 2016.\"

Final results

The translational nature of this project can have important implications for patients suffering from disorders of fear and anxiety and problems in social behavior such as found e.g. in Autism and Post-Traumatic Stress Disorder (PTSD) and for dysfunctional human social decision-making. Oxytocin and sex hormone receptors are currently most interesting candidates for current drug discovery targeting these behavioural domains. Our results showed pharmacological rescue of abnormal fear reactions by administering an oxytocin-receptor agonist. Further research in human fear and anxiety using this approach, combined with pharmacological manipulation, perhaps using selective oxytocin agonists that are under development, is thus warranted. Notably, safety or escape behaviors are maladaptive in many of disorders of fear and anxiety, and oxytocin receptors may well provide a treatment target.