A5056127593- Advanced Electron Microscopy Techniques and Applications
- Electron and X-Ray Spectroscopy Techniques
- Lipid Membrane Structure and Behavior
- Cellular transport and secretion
- Advanced Fluorescence Microscopy Techniques
- Force Microscopy Techniques and Applications
- Photosynthetic Processes and Mechanisms
- Neuroscience and Neuropharmacology Research
- Photoreceptor and optogenetics research
- Advanced X-ray Imaging Techniques
- Protein Structure and Dynamics
- Magnetic and Electromagnetic Effects
- Scientific Research and Discoveries
- Nuclear Structure and Function
- Ion channel regulation and function
- Cell Image Analysis Techniques
- Erythrocyte Function and Pathophysiology
- Machine Learning in Bioinformatics
- Gene Regulatory Network Analysis
- Characterization and Applications of Magnetic Nanoparticles
- Digital Holography and Microscopy
- Particle physics theoretical and experimental studies
- Toxin Mechanisms and Immunotoxins
- Neurological diseases and metabolism
- Neural Networks and Reservoir Computing
Max Planck Institute of Biochemistry
2015-2024
Max Planck Society
2004-2016
California Institute of Technology
2008
Northeastern University
1993-1995
The presynaptic terminal contains a complex network of filaments whose precise organization and functions are not yet understood. cryoelectron tomography experiments reported in this study indicate that these structures play prominent role synaptic vesicle release. Docked vesicles did make membrane to contact with the active zone but were instead linked it by tethers different length. Our observations consistent an exocytosis model which first anchored long (>5 nm) give way multiple...
Synucleins (α, β, γ-synuclein) are a family of abundant presynaptic proteins. α-Synuclein is causally linked to the pathogenesis Parkinson's disease (PD). In an effort define their physiological and pathological function or functions, we investigated effects deleting synucleins overexpressing α-synuclein PD mutations, in mice, on synapse architecture using electron microscopy (EM) cryoelectron tomography (cryo-ET). We show that regulators presynapse size synaptic vesicle (SV) pool...
Synaptic vesicles are embedded in a complex filamentous network at the presynaptic terminal. Before fusion, linked to active zone (AZ) by short filaments (tethers). The identity of molecules that form and regulate tethers remains unknown, but Rab3-interacting molecule (RIM) is prominent candidate, given its central role AZ organization. In this paper, we analyzed architecture RIM1α knockout (KO) mice cryo-electron tomography. stark contrast previous work on dehydrated, chemically fixed...
Synaptic vesicle tethering, priming, and neurotransmitter release require a coordinated action of multiple protein complexes. While physiological experiments, interaction data, structural studies purified systems were essential for our understanding the function individual complexes involved, they cannot resolve how actions integrate. We used cryo-electron tomography to simultaneously image presynaptic lipids at molecular resolution in their native composition, conformation, environment. Our...
Dopamine is an essential brain neuromodulator involved in reward and motor control. Dopaminergic (DA) neurons project to most areas, with particularly dense innervation the striatum. DA varicosities bind target striatal synapses form dopamine hub (DHS). However, basic features of release sites are still largely unknown. Here we studied ultrastructure fluorescent glutamatergic (GLU) synaptosomes isolated from striatum adult mice cryo-correlative light electron microscopy cryo-electron...
Synaptic transmission is characterized by fast, tightly coupled processes and complex signaling pathways that require a precise protein organization, such as the previously reported nanodomain colocalization of pre- postsynaptic proteins. Here, we used cryo-electron tomography to visualize synaptic complexes together with their native environment comprising interacting proteins lipids on 2- 4-nm scale. Using template-free detection classification, showed tripartite trans-synaptic assemblies...
Synaptic vesicle (SV) fusion with the plasma membrane (PM) proceeds through intermediate steps that remain poorly resolved. The effect of persistent high or low exocytosis activity on remains unknown. Using spray-mixing plunge-freezing cryo-electron tomography we observe events following synaptic stimulation at nanometer resolution in near-native samples. Our data suggest during stage immediately follows stimulation, termed early fusion, PM and SV curvature changes to establish a point...
Summary Cryo‐electron tomography allows three‐dimensional visualization of frozen‐hydrated, vitrified biological material at molecular resolution. Here, we summarize the most important sample preparation methods and technical aspects relevant for cryo‐electron tomography, as well its recent applications from isolated macromolecular complexes to entire cells tissues.
Many cellular processes depend on a precise structural organization of molecular components. Here, we established that neurons grown in culture provide suitable system for situ investigations structures by cryo-electron tomography, method allows high resolution, three-dimensional imaging fully hydrated, vitrified samples. A higher level detail components present our images allowed us to quantitatively characterize presynaptic and cytoskeletal organization, as well involved axonal transport...
By combining biochemical experiments with computer modelling of reactions we elucidated some the currently unresolved aspects calcium-calmodulin-dependent protein kinase II (CaMKII) activation and autophosphorylation that might be relevant for its physiological function provided a model incorporates in detail mechanism CaMKII at T286 is based on experimentally determined binding constants phosphorylation rates. To this end, developed detailed state available literature, constrained it data...
We describe comparative patch analysis for modeling the structures of multidomain proteins and protein complexes, apply it to PSD-95 protein. Comparative is a hybrid based on template complex docking, with greater applicability than higher accuracy docking. It relies structurally defined interactions each components, or their homologs, any other protein, irrespective its fold. For component, known binding modes fold are collected expanded by homologs. These then used restrain conventional...