TY  - JOUR
A1  - Pauli, Martin
A1  - Paul, Mila M.
A1  - Proppert, Sven
A1  - Mrestani, Achmed
A1  - Sharifi, Marzieh
A1  - Repp, Felix
A1  - Kürzinger, Lydia
A1  - Kollmannsberger, Philip
A1  - Sauer, Markus
A1  - Heckmann, Manfred
A1  - Sirén, Anna-Leena
T1  - Targeted volumetric single-molecule localization microscopy of defined presynaptic structures in brain sections
JF  - Communications Biology
N2  - Revealing the molecular organization of anatomically precisely defined brain regions is necessary for refined understanding of synaptic plasticity. Although three-dimensional (3D) single-molecule localization microscopy can provide the required resolution, imaging more than a few micrometers deep into tissue remains challenging. To quantify presynaptic active zones (AZ) of entire, large, conditional detonator hippocampal mossy fiber (MF) boutons with diameters as large as 10 mu m, we developed a method for targeted volumetric direct stochastic optical reconstruction microscopy (dSTORM). An optimized protocol for fast repeated axial scanning and efficient sequential labeling of the AZ scaffold Bassoon and membrane bound GFP with Alexa Fluor 647 enabled 3D-dSTORM imaging of 25 mu m thick mouse brain sections and assignment of AZs to specific neuronal substructures. Quantitative data analysis revealed large differences in Bassoon cluster size and density for distinct hippocampal regions with largest clusters in MF boutons. Pauli et al. develop targeted volumetric dSTORM in order to image large hippocampal mossy fiber boutons (MFBs) in brain slices. They can identify synaptic targets of individual MFBs and measured size and density of Bassoon clusters within individual untruncated MFBs at nanoscopic resolution.
KW  - mossy fiber synapses
KW  - CA3 pyrimidal cells
KW  - CA2+ channels
KW  - active zone
KW  - hippocampal
KW  - release
KW  - plasticity
KW  - proteins
KW  - platform
KW  - reveals
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259830
VL  - 4
ER  - 
TY  - JOUR
A1  - Heckmann, Manfred
A1  - Pauli, Martin
T1  - Visualizing presynaptic active zones and synaptic vesicles
JF  - Frontiers in Synaptic Neuroscience
N2  - The presynaptic active zone (AZ) of chemical synapses is a highly dynamic compartment where synaptic vesicle fusion and neurotransmitter release take place. During evolution the AZ was optimized for speed, accuracy, and reliability of chemical synaptic transmission in combination with miniaturization and plasticity. Single-molecule localization microscopy (SMLM) offers nanometer spatial resolution as well as information about copy number, localization, and orientation of proteins of interest in AZs. This type of imaging allows quantifications of activity dependent AZ reorganizations, e.g., in the context of presynaptic homeostatic potentiation. In combination with high-pressure freezing and optogenetic or electrical stimulation AZs can be imaged with millisecond temporal resolution during synaptic activity. Therefore SMLM allows the determination of key parameters in the complex spatial environment of AZs, necessary for next generation simulations of chemical synapses with realistic protein arrangements.
KW  - active zone
KW  - depression
KW  - facilitation
KW  - plasticity
KW  - potentiation
KW  - synapse
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-274687
SN  - 1663-3563
VL  - 14
ER  -