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Quantitative Nanoscale Imaging Of Synaptic Protein Organization
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Book Synopsis Quantitative Nanoscale Imaging of Synaptic Protein Organization by : Lara Laparra Cuervo
Download or read book Quantitative Nanoscale Imaging of Synaptic Protein Organization written by Lara Laparra Cuervo and published by . This book was released on 2019 with total page 172 pages. Available in PDF, EPUB and Kindle. Book excerpt: The arrival of super-resolution techniques has driven researchers to explore biological areas that were unreachable before. Such techniques not only allowed the improvement of spatial resolution in images but also the possibility to perform quantitative measurements at the single-molecule level. The interest in that particular field has been growing over the years and new and more sophisticated tools have been developed. Neuroscience has been one of the first fields to adapt and benefit from super-resolution microscopy. These techniques opened a new window of opportunity to reveal spatial organization of the neuronal cytoskeleton and the molecular organization and dynamics of the synapse, structures below the spatial resolution limit of conventional light microscopy. Protein organization and stoichiometry is central for synaptic transmission in neurons. Knowing the absolute numbers of proteins playing a key role in diseases can be of extreme interest in order to understand the mechanisms that lead to such neurological disorders. In this thesis we exploited the super-resolution techniques to develop a pioneering method for quantitative single-molecule measurements and to unravel the organization of a synaptic protein complex that was never visualized before with nanoscale spatial resolution. We developed a novel method in order to quantify the photoactivation efficiency of eight different photoswitchable fluorescent proteins commonly used in super-resolution experiments. We used the glycine receptor as a template because of its well-known stoichiometry and tagged eight different photoswitchable fluorescent proteins to the a- and ß-subunits of this receptor and transiently transfected them to Xenopus oocytes. The fact that the fluorescent proteins are genetically encoded make them highly suitable for quantitative single-molecule counting. The photoactivation efficiency, which is the percentage of a fluorescent protein that photoactivates into a fluorescently detectable form, plays a critical role in properly interpreting quantitative measurements. Moreover, we also focused our studies on super-resolution imaging of a synaptic protein complex, called LGI1 complex. This ensemble of proteins is one of the main key players involved in different neurological disorders. Leucine rich glioma activated 1 (LGI1) is a neuronal protein that forms a trans-synaptic bridge linking pre- and postsynaptic transmembrane proteins (ADAM22 and ADAM23) and helps to organize a multimeric complex at the synapse including AMPA receptors and voltage-gated potassium channels (VGKC). LGI1 autoimmune encephalitis is a severe neuropsychiatric disorder related to epilepsy where the patients produce autoantibodies against LGI1, which alter synaptic plasticity. However, the molecular mechanisms that lead to the observed problems in patients still remain largely unknown. Using well-characterized synaptic markers as molecular standards, we determined the positioning of LGI1 and the other related proteins within the synaptic space at nanoscale resolution by means of multi-color STORM. Further, the comparison of this molecular architecture in healthy neurons versus neurons treated with antibodies from patients suffering from LGI1 autoimmune encephalitis showed that these antibodies impact the nanoscale organization of pre-synaptic proteins. These results suggested a loss of LGI1 interaction with pre-synaptic proteins upon antibody binding and gave further insight into early changes in pathology.
Book Synopsis Nanoscale Imaging of Synapses by : U. Valentin Nägerl
Download or read book Nanoscale Imaging of Synapses written by U. Valentin Nägerl and published by Humana. This book was released on 2016-08-23 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Synapses underlie rapid and flexible neural communication in the brain and they hold the key to understanding higher brain functions in health and disease. Because they are very small and highly dynamic, it is very difficult to study them with traditional techniques. Fortunately, recent ground-breaking advances in optical microscopy (e.g. STED, PALM, STORM, SIM) have greatly improved our ability to image living synapses at the nanoscale, even down to the level of single molecules. The proposed volume brings together leading researchers to review these exciting new techniques and their application in neurobiological research. It will explain and discuss the basic principles behind the various superresolution modalities, how they are implemented, what their scope and limitations are etc. In addition, several key research discoveries on synapses enabled by these novel approaches will be highlighted.
Book Synopsis Quantitative Assessment of Synaptic Plasticity at the Molecular Scale with Multimodal Microscopy and Computational Tools by : Theresa Wiesner
Download or read book Quantitative Assessment of Synaptic Plasticity at the Molecular Scale with Multimodal Microscopy and Computational Tools written by Theresa Wiesner and published by . This book was released on 2021 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Learning and memory at the cellular and molecular levels are characterized by modulation of synaptic strength, involving the recruitment and re-localization of proteins within specific nanoscale synaptic domains. Most studies investigating the mechanisms of synaptic plasticity have been focussed on specific synapses, lacking a broad view of the diversity of synaptic changes in strength and protein re-organization across neural circuits. We use a combination of functional and super-resolution optical imaging in dissociated hippocampal cultures and artificial intelligence tools to classify the diversity of synapses, based on their functional and organizational characteristics. We measured synaptic activity using wide field microscopy to record miniature synaptic calcium transients (MSCTs) in neurons expressing the fluorescent calcium sensor GCaMP6f. We developed a deep learning approach to detect and segment these calcium events. Our results show that the amplitude and frequency of miniature calcium events are modulated by prior levels of circuit activity. In addition, we classified active synapses and identify differential recruitment of certain calcium dynamics depending on the plasticity paradigm used. To link the nanoscale organization of synaptic proteins with synaptic strength and plasticity, we optically resolved the organization of presynaptic (Bassoon, RIM1/2) and postsynaptic (PSD95, Homer1c) scaffolding proteins using STED (Stimulated Emission Depletion) nanoscopy. Using Python-based statistical object distance analysis (pySODA), we show that plasticity-inducing stimuli differentially alter the spatial organization of these proteins. In particular, PSD95 and Bassoon proteins show a treatment-dependent change in organization, associated either with synaptic potentiation or depression. Furthermore, using unsupervised machine learning approaches, we reveal the rich diversity of synaptic protein subtypes exhibiting differential remodeling. To investigate further the link between synaptic protein architecture and synaptic function, we aimed to combine functional and super-resolution imaging. We therefore used a machine learning approach to optimize live-cell imaging parameters for time-lapse imaging and combined this with the optimization of glutamate uncaging parameters to probe corresponding calcium signals. Our approach allows to characterize the population of synapses in terms of their activity rate and synaptic protein organization, providing a basis for further exploring the diverse molecular mechanisms of synaptic plasticity.
Book Synopsis Quantifying and Controlling the Nano-Architecture of Neuronal Synapses by : Xiaobing Chen
Download or read book Quantifying and Controlling the Nano-Architecture of Neuronal Synapses written by Xiaobing Chen and published by Frontiers Media SA. This book was released on 2022-09-27 with total page 195 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Advanced Optical Methods for Brain Imaging by : Fu-Jen Kao
Download or read book Advanced Optical Methods for Brain Imaging written by Fu-Jen Kao and published by Springer. This book was released on 2018-05-14 with total page 342 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book highlights the rapidly developing field of advanced optical methods for structural and functional brain imaging. As is known, the brain is the most poorly understood organ of a living body. It is indeed the most complex structure in the known universe and, thus, mapping of the brain has become one of the most exciting frontlines of contemporary research. Starting from the fundamentals of the brain, neurons and synapses, this book presents a streamlined and focused coverage of the core principles, theoretical and experimental approaches, and state-of-the-art applications of most of the currently used imaging methods in brain research. It presents contributions from international leaders on different photonics-based brain imaging modalities and techniques. Included are comprehensive descriptions of many of the technology driven spectacular advances made over the past few years that have allowed novel insights of the structural and functional details of neurons. The book is targeted at researchers, engineers and scientists who are working in the field of brain imaging, neuroscience and connectomics. Although this book is not intended to serve as a textbook, it will appeal to undergraduate students engaged in the specialization of brain imaging.
Book Synopsis Springer Handbook of Microscopy by : Peter W. Hawkes
Download or read book Springer Handbook of Microscopy written by Peter W. Hawkes and published by Springer Nature. This book was released on 2019-11-02 with total page 1561 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book features reviews by leading experts on the methods and applications of modern forms of microscopy. The recent awards of Nobel Prizes awarded for super-resolution optical microscopy and cryo-electron microscopy have demonstrated the rich scientific opportunities for research in novel microscopies. Earlier Nobel Prizes for electron microscopy (the instrument itself and applications to biology), scanning probe microscopy and holography are a reminder of the central role of microscopy in modern science, from the study of nanostructures in materials science, physics and chemistry to structural biology. Separate chapters are devoted to confocal, fluorescent and related novel optical microscopies, coherent diffractive imaging, scanning probe microscopy, transmission electron microscopy in all its modes from aberration corrected and analytical to in-situ and time-resolved, low energy electron microscopy, photoelectron microscopy, cryo-electron microscopy in biology, and also ion microscopy. In addition to serving as an essential reference for researchers and teachers in the fields such as materials science, condensed matter physics, solid-state chemistry, structural biology and the molecular sciences generally, the Springer Handbook of Microscopy is a unified, coherent and pedagogically attractive text for advanced students who need an authoritative yet accessible guide to the science and practice of microscopy.
Book Synopsis Shedding Light on the Nervous System: Progress in Neurophotonics Research by : Jean-Claude Béïque
Download or read book Shedding Light on the Nervous System: Progress in Neurophotonics Research written by Jean-Claude Béïque and published by Frontiers Media SA. This book was released on 2022-07-01 with total page 250 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Imaging Synapse Structure and Function by : George J. Augustine
Download or read book Imaging Synapse Structure and Function written by George J. Augustine and published by Frontiers Media SA. This book was released on 2017-07-06 with total page 127 pages. Available in PDF, EPUB and Kindle. Book excerpt: Development of new imaging technologies in recent years has transformed neuroscience in profound ways. Following on the heels of the revolution based on the Green Fluorescent Protein, refined genetically-encoded fluorescent reporters and genetic targeting strategies now enable optical recording of synaptic transmission in defined neuronal populations at speeds approaching the enviable temporal resolution of electrophysiology. Super-resolution light microscopy permits observation of synapses and their molecular machinery at sub-diffraction resolution. At the ultrastructural level, automated forms of electron microscopy, improvements in specimen fixation methods, and recent efforts to correlate data from light and electron micrographs now make the reconstruction of functional neural circuits a reality. Finally, the use of optogenetic actuators, such as channelrhodopsins, allows precise temporal and spatial manipulation of neuronal activity and is revealing profound insights into the organization of neural circuits and their roles in behavior. This research topic highlights recent advances in both light and electron microscopy, with a specific focus on approaches that combine innovations from several different fields to obtain novel information about synapse structure and function. We are confident that this collection of articles - three original research papers, six reviews, one methods paper and one perspective article - will enable neuroscientists to achieve the next generation of experiments aimed at cracking the neural code.
Book Synopsis Translational Research Methods in Neurodevelopmental Disorders by : Stéphane Martin
Download or read book Translational Research Methods in Neurodevelopmental Disorders written by Stéphane Martin and published by Springer Nature. This book was released on 2022-09-29 with total page 271 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume explores the latest techniques used to study neurodevelopmental diseases (NDD) that range from molecular aspects to integrated research approaches and brain imaging in living rodents. Chapters in this book cover topics such as protocols to deliver shRNA in vitro and in vivo using lentiviral particles to knock-down specific protein expression; experimental procedures to use recombinant fluorescent probes to visualize endogenous proteins at the mammalian synapse; CRISPR/Cas9 toolkit to assess either gain- or loss-of-gene function in brain organoids; detailed protocols to use in vivo manipulations to correct the behavioral phenotypes in cognitive disorder mouse lines; and experimental approaches to genetically engineer macaque models of NDDs and investigate how genetic predisposition may cause neural and functional alterations. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and practical, Translational Research Methods in Neurodevelopmental Disorders is a valuable resource for all researchers who want to learn more about this important and developing field.
Book Synopsis Emerging Mechanisms in Dynamic GABAergic Inhibition by : Shiva K. Tyagarajan
Download or read book Emerging Mechanisms in Dynamic GABAergic Inhibition written by Shiva K. Tyagarajan and published by Frontiers Media SA. This book was released on 2021-12-01 with total page 171 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Super-Resolution Imaging in Biomedicine by : Alberto Diaspro
Download or read book Super-Resolution Imaging in Biomedicine written by Alberto Diaspro and published by CRC Press. This book was released on 2016-11-03 with total page 448 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book encompasses the full breadth of the super-resolution imaging field, representing modern techniques that exceed the traditional diffraction limit, thereby opening up new applications in biomedicine. It shows readers how to use the new tools to increase resolution in sub-nanometer-scale images of living cells and tissue, which leads to new information about molecules, pathways and dynamics. The book highlights the advantages and disadvantages of the techniques, and gives state-of-the-art examples of applications using microscopes currently available on the market. It covers key techniques such as stimulated emission depletion (STED), structured illumination microscopy (SSIM), photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM). It will be a useful reference for biomedical researchers who want to work with super-resolution imaging, learn the proper technique for their application, and simultaneously obtain a solid footing in other techniques.
Book Synopsis Cellular and Molecular Neurophysiology by : Constance Hammond
Download or read book Cellular and Molecular Neurophysiology written by Constance Hammond and published by Elsevier. This book was released on 2024-06-03 with total page 602 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cellular and Molecular Neurophysiology, Fifth Edition is the only up-to-date textbook on the market that focuses on the molecular and cellular physiology of neurons and synapses. Hypothesis-driven rather than a dry presentation of the facts, the book promotes a real understanding of the function of nerve cells that is useful for practicing neurophysiologists and students in graduate-level courses on the topic alike. This new edition explains the molecular properties and functions of excitable cells in detail and teaches students how to construct and conduct intelligent research experiments. The content is firmly based on numerous experiments performed by top experts in the field. The new edition contains new chapters on recording neuronal activity, iconotrophic and metabotropic receptors for sensory transduction, and a section containing exercises for further learning. This book will be a useful resource for neurophysiologists, neurobiologists, neurologists, and students taking graduate-level courses on neurophysiology. Authoritative foundational coverage of basic cellular and molecular neurophysiology Includes new chapters on recording neuronal activity, iconotrophic and metabotropic receptors for sensory transduction Provides fifteen appendices that describe how neurobiological techniques are interspersed in the text Presents enhanced coverage of new methodologies and experimental techniques
Book Synopsis Synapse Development and Maturation by : Pasko Rakic
Download or read book Synapse Development and Maturation written by Pasko Rakic and published by Academic Press. This book was released on 2020-05-29 with total page 560 pages. Available in PDF, EPUB and Kindle. Book excerpt: Synapse Development and Maturation, the latest release in the Comprehensive Developmental Neuroscience series, presents the latest information on the genetic, molecular and cellular mechanisms of neural development. The book provides a much-needed update that underscores the latest research in this rapidly evolving field, with new section editors discussing the technological advances that are enabling the pursuit of new research on brain development. This volume focuses on the synaptogenesis and developmental sequences in the maturation of intrinsic and synapse-driven patterns. Features leading experts in various subfields as section editors and article authors Presents articles that have been peer reviewed to ensure accuracy, thoroughness and scholarship Includes coverage of mechanisms which regulate synapse formation and maintenance during development Covers neural activity, from cell-intrinsic maturation, to early correlated patterns of activity
Book Synopsis Frontiers in Synaptic Neuroscience - Editor's Pick 2021 by : P. Jesper Sjöström
Download or read book Frontiers in Synaptic Neuroscience - Editor's Pick 2021 written by P. Jesper Sjöström and published by Frontiers Media SA. This book was released on 2021-07-21 with total page 168 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Single Molecule Microscopy in Neurobiology by : Nobuhiko Yamamoto
Download or read book Single Molecule Microscopy in Neurobiology written by Nobuhiko Yamamoto and published by Humana. This book was released on 2021-05-13 with total page 331 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume looks at the methodology and techniques used by experts to study how certain molecules function in specific locations, and their temporal patterns. Chapters in this book cover topics such as in vivo single-molecule tracking of voltage-gated calcium channels with split-fluorescent proteins in CRISPR-engineering C. elegans; protein-protein interactions in membranes using single particle tracking; neuropathological diseases revealed by quantum-dot single particle tracking; SPoD-OnSPAN; and investigating molecular diffusion inside small neuronal compartments with two-photon fluorescence correlation spectroscopy. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and comprehensive, Single Molecule Microscopy is a valuable resource for any researcher interested in learning more about this important field.
Book Synopsis Methods for Synaptic Interrogation by : Dirk Feldmeyer
Download or read book Methods for Synaptic Interrogation written by Dirk Feldmeyer and published by Frontiers Media SA. This book was released on 2020-07-14 with total page 276 pages. Available in PDF, EPUB and Kindle. Book excerpt: This eBook is a collection of articles from a Frontiers Research Topic. Frontiers Research Topics are very popular trademarks of the Frontiers Journals Series: they are collections of at least ten articles, all centered on a particular subject. With their unique mix of varied contributions from Original Research to Review Articles, Frontiers Research Topics unify the most influential researchers, the latest key findings and historical advances in a hot research area! Find out more on how to host your own Frontiers Research Topic or contribute to one as an author by contacting the Frontiers Editorial Office: frontiersin.org/about/contact.
Book Synopsis Molecular Nanomachines of the Presynaptic Terminal, Volume II by : Silvio O. Rizzoli
Download or read book Molecular Nanomachines of the Presynaptic Terminal, Volume II written by Silvio O. Rizzoli and published by Frontiers Media SA. This book was released on 2022-06-30 with total page 168 pages. Available in PDF, EPUB and Kindle. Book excerpt:
