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BP: Fachverband Biologische Physik
BP 8: Poster Session Ia
BP 8.12: Poster
Monday, March 18, 2024, 18:00–20:30, Poster C
Interactions between synaptic vesicles and cytoskeletal filaments — •Tiago Mimoso1, Rajdeep Chowdhury2, Saheli Dey1, Christian Hoffmann3, Dragomir Milovanovic3, Silvio Rizzoli2, and Sarah Köster1 — 1Institute for X-Ray Physics, University of Göttingen, Germany — 2Institute for Neuro- and Sensory Physiol- ogy, University Medical Center Göttingen, Germany — 3Laboratory of Molecular Neuroscience, DZNE, Germany
Signal transmission of neurons occurs both electrically and chemically. The chemical signal is transported by synaptic vesicles (SVs) via the synaptic cleft to an adjacent neuron. Thus, these SVs are found in the synapse, within the so-called synaptic bouton. Here, the SVs are surrounded by cytoskeletal filaments, including dynamic microtubules (MTs) that undergo rapid assembly and disassembly. Some studies suggest interactions between SVs and the cytoskeletal filaments. Therefore, we now ask the question of what influence the presence of SVs has on microtubules. We employ a reconstituted in vitro system, by attaching the SVs to the surface and imaging the dynamic microtubules by total internal reflection fluorescence microscopy to obtain the growth rate, disassembly rate, catastrophe frequency and rescue frequency. We present an approach for attaching SVs to a surface using protein G and a primary antibody targeting a membrane protein in SVs. The MT are attached using biotin-neutravidin complex. This method grants control over the SVs and MT positioning, enabling a comprehensive study of their dynamic interactions.
Keywords: Synaptic Vesicles; Microtubule; TIRFM; in vitro