Plasma membrane probes for photo-controlled release of signaling molecules
The cell plasma membrane (PM) of eukaryotic cells is a highly dynamic structure, which plays important role in diverse biological processes such as cellular uptake, neural communication, muscle contraction, and cell trafficking an...
The cell plasma membrane (PM) of eukaryotic cells is a highly dynamic structure, which plays important role in diverse biological processes such as cellular uptake, neural communication, muscle contraction, and cell trafficking and signaling. The PMs are also responsible for communication between cells, especially between neurons. In particular, the exocytosis process leads to liberation of neurotransmitters and neuromodulators. On the other hand, it is a platform for receptors that convert the recognition of neuroactive molecules into ion flux and/or action potential. Therefore, to modulate and monitor the activity of neurons, an attractive approach is to design molecular tools that target cell PMs. In this project, we will combine PM-targeting anchors with photoactive fluorescent cages capable to photo-release signaling molecules. The obtained chemical tools (MemCages) will be applied for 1) imaging the neuronal organization and 2) photo-controlled activation of neurons and brain of mice by photo-uncaging of signaling molecules close to PM. The work is composed of 3 steps. First is synthesis of MemCages for dopamine and glutamate and their validation in solvents and model lipid membranes. Second is application of MemCages in cellular systems, which includes imaging and photo-stimulation of model cancer cell lines and neurons. The first step will be in vivo validation of MemCages, which will be done during the secondment. The applicant and the host laboratory plan extensive two-way knowledge transfer: the applicant will gain knowledge and expertise in membrane probe design, bioimaging in model membranes, cells and animals, whereas he will provide his expertise on design and characterization of molecular photocages for signaling molecules. The successful utility of the developed chemical tools could open new avenues for the biologists and neuroscientists working in the field of cell signaling, neuroscience and diagnosis and prevention of neurological disorders.ver más
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