@misc{18296,
  abstract     = {It is widely believed that information storage in neuronal circuits involves nanoscopic structural changes at synapses, resulting in the formation of synaptic engrams. However, direct evidence for this hypothesis is lacking. To test this conjecture, we combined chemical potentiation, functional analysis by paired pre-postsynaptic recordings, and structural analysis by electron microscopy (EM) and freeze-fracture replica labeling (FRL) at the murine hippocampal mossy fiber synapse, a key synapse in the trisynaptic circuit of the hippocampus. Biophysical analysis of synaptic transmission revealed that forskolin-induced chemical potentiation increased the readily releasable vesicle pool size and vesicular release probability by 146% and 49%, respectively. Structural analysis of mossy fiber synapses by EM and FRL demonstrated an increase in the number of vesicles close to the plasma membrane and the number of clusters of the priming protein Munc13-1, indicating an increase in the number of both docked and primed vesicles. Furthermore, FRL analysis revealed a significant reduction of the distance between Munc13-1 and CaV2.1 Ca2+ channels, suggesting reconfiguration of the channel-vesicle coupling nanotopography. Our results indicate that presynaptic plasticity is associated with structural reorganization of active zones. We propose that changes in potential nanoscopic organization at synaptic vesicle release sites may be correlates of learning and memory at a plastic central synapse.},
  author       = {Kim, Olena},
  keywords     = {Hippocampal mossy fiber synapses, short-term potentiation, long-term potentiation, presynaptic plasticity, electron microscopy, freeze-fracture replica labeling, paired recordings, forskolin, cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), neuromodulation, synaptic vesicle pools, presynaptic Ca2+ channels, Munc13, docking, priming, active zone},
  publisher    = {Institute of Science and Technology Austria},
  title        = {{Presynaptic cAMP-PKA-mediated potentiation induces reconfiguration of synaptic vesicle pools and channel-vesicle coupling at hippocampal mossy fiber boutons}},
  doi          = {10.15479/AT:ISTA:18296},
  year         = {2024},
}

@inbook{9756,
  abstract     = {High-resolution visualization and quantification of membrane proteins contribute to the understanding of their functions and the roles they play in physiological and pathological conditions. Sodium dodecyl sulfate-digested freeze-fracture replica labeling (SDS-FRL) is a powerful electron microscopy method to study quantitatively the two-dimensional distribution of transmembrane proteins and their tightly associated proteins. During treatment with SDS, intracellular organelles and proteins not anchored to the replica are dissolved, whereas integral membrane proteins captured and stabilized by carbon/platinum deposition remain on the replica. Their intra- and extracellular domains become exposed on the surface of the replica, facilitating the accessibility of antibodies and, therefore, providing higher labeling efficiency than those obtained with other immunoelectron microscopy techniques. In this chapter, we describe the protocols of SDS-FRL adapted for mammalian brain samples, and optimization of the SDS treatment to increase the labeling efficiency for quantification of Cav2.1, the alpha subunit of P/Q-type voltage-dependent calcium channels utilizing deep learning algorithms.},
  author       = {Kaufmann, Walter and Kleindienst, David and Harada, Harumi and Shigemoto, Ryuichi},
  booktitle    = { Receptor and Ion Channel Detection in the Brain},
  isbn         = {9781071615218},
  keywords     = {Freeze-fracture replica: Deep learning, Immunogold labeling, Integral membrane protein, Electron microscopy},
  pages        = {267--283},
  publisher    = {Humana},
  title        = {{High-Resolution localization and quantitation of membrane proteins by SDS-digested freeze-fracture replica labeling (SDS-FRL)}},
  doi          = {10.1007/978-1-0716-1522-5_19},
  volume       = {169},
  year         = {2021},
}

@article{8586,
  abstract     = {Cryo-electron microscopy (cryo-EM) of cellular specimens provides insights into biological processes and structures within a native context. However, a major challenge still lies in the efficient and reproducible preparation of adherent cells for subsequent cryo-EM analysis. This is due to the sensitivity of many cellular specimens to the varying seeding and culturing conditions required for EM experiments, the often limited amount of cellular material and also the fragility of EM grids and their substrate. Here, we present low-cost and reusable 3D printed grid holders, designed to improve specimen preparation when culturing challenging cellular samples directly on grids. The described grid holders increase cell culture reproducibility and throughput, and reduce the resources required for cell culturing. We show that grid holders can be integrated into various cryo-EM workflows, including micro-patterning approaches to control cell seeding on grids, and for generating samples for cryo-focused ion beam milling and cryo-electron tomography experiments. Their adaptable design allows for the generation of specialized grid holders customized to a large variety of applications.},
  author       = {Fäßler, Florian and Zens, Bettina and Hauschild, Robert and Schur, Florian KM},
  issn         = {1047-8477},
  journal      = {Journal of Structural Biology},
  keywords     = {electron microscopy, cryo-EM, EM sample preparation, 3D printing, cell culture},
  number       = {3},
  publisher    = {Elsevier},
  title        = {{3D printed cell culture grid holders for improved cellular specimen preparation in cryo-electron microscopy}},
  doi          = {10.1016/j.jsb.2020.107633},
  volume       = {212},
  year         = {2020},
}

@inproceedings{11222,
  author       = {Kim, Olena and Borges Merjane, Carolina and Jonas, Peter M},
  booktitle    = {Intrinsic Activity},
  issn         = {2309-8503},
  keywords     = {hippocampus, mossy fibers, readily releasable pool, electron microscopy},
  location     = {Innsbruck, Austria},
  number       = {Suppl. 1},
  publisher    = {Austrian Pharmacological Society},
  title        = {{Functional analysis of the docked vesicle pool in hippocampal mossy fiber terminals by electron microscopy}},
  doi          = {10.25006/ia.7.s1-a3.27},
  volume       = {7},
  year         = {2019},
}

@article{21518,
  abstract     = {The present investigation confirms that initially implemented procedure to produce poly(methylidene malonate 2.1.2) (PMM 2.1.2) nanoparticles (Lescure et al. Pharm Res 1994;11:1270–77) lead to products mostly containing plasticizing oligomers which strongly lowered glass-transition temperature (Tg), dramatically reduced nanoparticle consistency and rendered them too sensitive to solubilization when diluted in an aqueous medium. From MALDI-TOF spectroscopy analysis, performed on intact colloids, emerged some structural information about these oligomeric species which could result from an intramolecular cyclization mechanism occurring soon in the course of the polymerization process. Thus, with the objective of overcoming these drawbacks, this contribution deals with the variations of manufacturing specifications such as pH and magnetic stirring speed to try and modulate molecular weight (Mw) of nanoparticle constituents and reduce oligomer concentration. Although the analyses performed on these new nanoparticles were rather encouraging, the colloid formation yield became so low that it required the developement of other methodologies, excluding a previous emulsion step, and allowing a controlled production of PMM 2.1.2-made nanoparticles having better physico-chemical characteristics while keeping good pharmaceutical capabilities.},
  author       = {Breton, P and Guillon, X and Roy, D and Lescure, F and Riess, G and Bru, N and Roques-Carmes, Charles},
  issn         = {0142-9612},
  journal      = {Biomaterials},
  keywords     = {Colloid physico-chemical analysis, Colloidal drug carriers, MALDI-TOF spectroscopy, Nanoparticles, Poly(methylidene malonate), Scanning electron microscopy},
  number       = {1-3},
  pages        = {271--281},
  publisher    = {Elsevier},
  title        = {{Physico-chemical characterization, preparation and performance of poly (methylidene malonate 2.1.2) nanoparticles}},
  doi          = {10.1016/s0142-9612(97)00243-3},
  volume       = {19},
  year         = {1998},
}