The underlying goal of neuroscience research is to understand how the nervous system functions to bring about behavior. A detailed map of neural circuits is required for scientists to tackle this question. To this purpose, we developed a synthetic and genetically-encoded system, TRanscellular ACtivation of Transcription (TRACT) to monitor cell-cell contact. Upon ligand-receptor interaction at sites of cell-cell contact, the transmembrane domain of an engineered Notch receptor is cleaved by intramembrane proteolysis and releases a fragment that regulates transcription in the receptor-expressing cell. We demonstrate that in cultured cells, the synthetic receptor can be activated to drive reporter gene expression by co-incubation with ligand-expressing cell or by growth on ligand-coated surfaces. We further show that TRACT can detect interactions between neurons and glia in the Drosophila brain; expressing the ligand in spatially-restricted subsets of neurons leads to transcription of a reporter in the glial cells that interact with those neurons. To optimize TRACT for neural tracing, we attempted to target the synthetic receptor to post-synaptic sites by fusion with the intracellular domain of Drosophila neuroligin2. However, this modification only facilitate the receptor to be localized homogeneously throughout the neurites. The induction data of the modified receptor shows that the new receptor has better sensitivity compared to the original receptor, but the ligand-receptor interaction still happened at non-synaptic sites of membrane contact. To further target the ligand to pre-synaptic sites, we fused the ligand to different pre-synaptic markers. We found the one fused with synaptobrevin is likely located at axon terminals, but only able to trigger moderate induction. Therefore, more examinations are required to further characterize the capability of this ligand. In summary, TRACT is useful for monitoring cell-cell interactions in animals and could also be used to genetically manipulate cells based on contact. Moreover, we believe that proper targeting of the ligand to synaptic sites will improve the specificity of TRACT for synaptic connections in the future.

Les cellules dendritiques plasmacytoides (pDCs), spécialisées dans la réponse antivirale, produisent de fortes quantités d’interféron (IFN) lorsqu’elles sont en contact avec des cellules infectées par des virus. Pourtant, les pDCs sont réfractaires à l’infection. Ce mécanisme d’activation de la réponse antivirale par le contact physique avec les cellules infectées, nouvellement découvert, constituerait un aspect général des voies de défense de l’hôte contre les virus.En utilisant le virus de l’Hépatite C et de la Dengue comme modèle viral, nous avons observé une réorganisation moléculaire au niveau des contacts entre les pDCs et les cellules infectées. La polarisation d’éléments cellulaires, notamment de régulateurs du cytosquelette d’actine et de molécules de la machinerie d’endocytose en direction du contact favoriserait son établissement et/ou sa stabilisation ainsi qu’une transmission efficace d’éléments viraux, ensuite reconnus par les pDCs. Nous avons également démontré que les pDCs effectuent des contacts plus stables et présentent une polarisation plus importante d’éléments cellulaires aux contacts avec des cellules infectées qu’avec des cellules non infectées. Ces interactions présentent des similarités avec les synapses, contacts cellulaires organisés impliqués dans la communication cellulaire. Notamment, les synapses immunologiques jouent un rôle important dans l’activation de la réponse immunitaire adaptative. Nous proposons donc de nommer ces contacts activateurs de pDCs des « synapses immunologiques innées ». Ce mécanisme représenterait un processus de reconnaissance des infections par les pDCs généralisable à différents types de virus, par « scan » du statut infectieux des cellules par contact. Nos résultats suggèrent également que des éléments viraux s’accumulent au niveau de ces contacts. Ces éléments diffèrent en fonction du type d’infection. Notamment, nous avons mis en évidence dans un contexte d’infection par le virus de la Dengue que des structures virales non canoniques et non infectieuses, différentes des particules virales infectieuses dites « classiques », jouent un rôle important dans l’activation de la réponse antivirale. Notre travail apporte un nouvel angle d’analyse de l’activation des pDCs et des stratégies de détection des infections virales par l’hôte. / Plasmacytoid dendritic cells (pDCs), specialized in the antiviral response, are important producers of interferons (IFN) after cell-cell contacts with virally infected cells. Nonetheless, they are poorly permissive to the majority of viral infections. This newly uncovered mechanism of the activation of an antiviral response by physical cell-cell contacts with infected cells could constitute a general aspect of the host defense against viral infections.Using Hepatitis C virus and Dengue virus as models, we observed a molecular reorganization of the contacts between pDCs and infected cells. The polarization toward contacts of cellular elements, such as regulators of the actin cytoskeleton and components of the endocytic machinery could favor their establishment and/or their stabilization, as well as the efficient transmission of viral elements that are recognized by pDCs. We also demonstrated that pDCs contacts with infected cells are more stable and present a higher polarization of cellular components than contacts with uninfected cells. These interactions present similarities with synapses, a type of organized contact involved in cell-to-cell communication. Notably, immunological synapses are known to play an important role in the activation of the adaptive immune response. We thus propose to call these pDC-activating contacts « innate immunological synapses ». This mechanism could represent a general process of recognition of viral infections by pDCs, by « scanning » the infectious status of the cells by cell-cell contacts. Our results also suggest that viral elements cluster at the level of contacts. These elements differ depending on the type of viral infection. Notably, we observed in the context of Dengue virus infection that non-infectious non-canonical viral structures, that differ from the « classical » viral infectious particles, play an important role in the activation of the antiviral response. Our work brings a new light in the mechanisms of pDC activation and in the host defense strategies against viral infection.