Agonist - G protein-coupled receptor interactions, recycling and cell-surface targeting in neuroendocrine systems

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My research program is geared towards the elucidation of the mechanisms regulating cell-surface targeting and sub-cellular localization of G protein-coupled receptors (GPCRs) in neuroendocrine cells and neurons. Both these processes are key to the normal functioning of cells, in particular of nerve cells which due to their complexity face particular challenges in maintaining correct intracellular targeting and positioning of their key component proteins. To this end we use classical and cutting-edge cell biological techniques and collaborate with laboratories to create genetically modified variants of cells and laboratory mice by the new technique of genome editing using CRISPR/Cas9 technology. Moreover, the recent advent of new microscopic techniques surmounting the resolution limit of light microscopy based on the clever exploitation of the photo-chemical properties of many fluorophores, referred to as super-resolution microscopy (SRM), promises the direct visualization of protein interactions regulating these processes. Much like transmission electron microscopy, the new SRM techniques have specific requirements for sample preparation and data analysis requiring substantial research to establish suitable experimental protocols and intense training of HQP to perform the procedures. Thanks to the access to a SRM microscope that is based on the point-localization, bi-plane principle of SRM provided by the MNI Microscopy Core Facility, we recently visualized for instance the sub-cellular localization of sst2A receptors at clusters of gephyrin, a hallmark structural component of the post-synaptic density at inhibitory synapses in the central nervous system at 50 nm resolution. This is at least 5 times better than the best standard light microscope can do and approaches the size of individual protein molecules. Therefore, the present application builds on my expertise in super-resolution microscopy and the cell biology of GPCRs to further my research and training program by a) integrating the new SRM techniques with established protocols in cell biology, confocal and electron microscopy and b) using all of these approaches to elucidate the open questions around the regulation of intracellular trafficking and targeting of GPCRs in neuroendocrine and neuronal cells.