During this period, the researcher was able to perform the six work packages (WP) included in the two objectives described above. In the first WP, she was able to perform behavioural tests under acute and chronic stress. She could also measure mitochondrial calcium dynamics ex vivo, in vitro and in vivo (2nd WP), after corticosterone treatment and acute stress. The principal investigator could also measure ATP, mitochondrial respiration and ROS production by high-resolution respirometry. MAMs were also measured from acutely stressed animals. Protein expression of several MAMs-localized proteins involved in calcium transfer was also evaluated by qPCR in chronic stress samples. During the completion of this WP, the investigator received trainings related to dissection methods, OROBOROS, confocal trainings and other techniques necessary to complete the experiments included in the appendix 1 of the project. The third WP of the project included the set up and proteomics of MAMs in a region and cell-specific manner. During the period of the action the main investigator was able to design and produce the constructs and test them in vitro and in vivo, optimizing the amount of biotinylated proteins detected in the NAc. Further experiments are in progress. For the WP 4, the investigator worked with the Mfn2 flox/flox line to knocked out the Mfn2 specifically in D1 neurons and test the impact in social dominance after the acute stress. The main results obtained during the development of this project were that the correlation of corticosterone with mitochondrial respiration in NAc was lost under acute stress and inversely correlated in the mPFC under chronic stress. Acute stress produced metabolic stress in NAc, affecting ATP production that correlated with animal performance in the tube test. These corticosterone-mediated effects also differently affected MAMs in the two types of medium spiny neurons in the NAc. These results were reported and disseminated in several congresses where the investigator presented her work as a poster. She also participated in several dissemination activities. The researcher will publish the main results after the end of all experimentation duly acknowledging the EU and specifying the number of the action. In case the experimenter finds other results that can be exploited in another way, they will be exploited by applying for a patent with the help of the EPFL Technology Transfer Office, as specified in the grant agreement of the action. During the duration of the action, the researcher already worked on her professional development plan, participating in mentorship programs. The researcher applied earlier this year for the Serra Hunter program to be eligible for a tenure-track position in her home country. In parallel, she is training, collaborating, networking and pursuing other options for positions in industry in Switzerland. In addition, other competencies that the researcher developed during the course of the action were the opportunity to supervise two bachelor students and training in license writing. After writing and being interviewed she obtained a Robert Gnehm grant to hire a technician for her project; concurrently, she also obtained a FENS/IBRO society travel grant. She collaborated on two scientific publications with laboratory and external colleagues during the period of the action. The researcher attended several courses on technical data management and transfer to industry and during the period of the action and currently, she is attending French courses to improve her communication skills and job opportunities.
