Three‐dimensional organization of the cytoskeleton: A cryo‐electron tomography perspective

Chakraborty, Saikat; Jasnin, Marion; Baumeister, Wolfgang

doi:10.1002/pro.3858

Cited by 28 publications

(21 citation statements)

References 184 publications

(296 reference statements)

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“…Additionally, focused-ion beam (FIB) milling instrumentation allows for exquisitely thin and detailed cell sections to be isolated ( Figure 6A) (163,164). Cryo-ET examples include cytosolic and mitochondrial structures of actively translating ribosomes (165), complex actin and microtubule network assembly (166,167), and intriguing views of the neural synapse (168). To examine the nuclear pore complex, detergent solubilization and removal of nucleic acids has enabled thinning samples as much as possible while maintaining 3D structure.…”

Section: Future Techniques For Understanding the Ismentioning

confidence: 99%

Considerations of Antibody Geometric Constraints on NK Cell Antibody Dependent Cellular Cytotoxicity

Murin

2020

Front. Immunol.

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It has been well-established that antibody isotype, glycosylation, and epitope all play roles in the process of antibody dependent cellular cytotoxicity (ADCC). For natural killer (NK) cells, these phenotypes are linked to cellular activation through interaction with the IgG receptor FcγRIIIa, a single pass transmembrane receptor that participates in cytoplasmic signaling complexes. Therefore, it has been hypothesized that there may be underlying spatial and geometric principles that guide proper assembly of an activation complex within the NK cell immune synapse. Further, synergy of antibody phenotypic properties as well as allosteric changes upon antigen binding may also play an as-of-yet unknown role in ADCC. Understanding these facets, however, remains hampered by difficulties associated with studying immune synapse dynamics using classical approaches. In this review, I will discuss relevant NK cell biology related to ADCC, including the structural biology of Fc gamma receptors, and how the dynamics of the NK cell immune synapse are being studied using innovative microscopy techniques. I will provide examples from the literature demonstrating the effects of spatial and geometric constraints on the T cell receptor complex and how this relates to intracellular signaling and the molecular nature of lymphocyte activation complexes, including those of NK cells. Finally, I will examine how the integration of high-throughput and "omics" technologies will influence basic NK cell biology research moving forward. Overall, the goal of this review is to lay a basis for understanding the development of drugs and therapeutic antibodies aimed at augmenting appropriate NK cell ADCC activity in patients being treated for a wide range of illnesses.

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Section: Future Techniques For Understanding the Ismentioning

confidence: 99%

Considerations of Antibody Geometric Constraints on NK Cell Antibody Dependent Cellular Cytotoxicity

Murin

2020

Front. Immunol.

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“…Classical EM techniques had clear limitations with respect to efficiency and the use of fixation and staining methods that might not always represent axonal structure in all accuracy. New techniques such as large-volume EM or cryo-electron tomography ( Kubota et al, 2018 ; Xu et al, 2017 ; Chakraborty et al, 2020 ) provide ever-improving means to drive the discovery processes toward good understanding of axon structure, which will eventually have important implications for work on neuronal pathologies.…”

Section: Introductionmentioning

confidence: 99%

Cytoskeletal organization of axons in vertebrates and invertebrates

Prokop

2020

Journal of Cell Biology

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The maintenance of axons for the lifetime of an organism requires an axonal cytoskeleton that is robust but also flexible to adapt to mechanical challenges and to support plastic changes of axon morphology. Furthermore, cytoskeletal organization has to adapt to axons of dramatically different dimensions, and to their compartment-specific requirements in the axon initial segment, in the axon shaft, at synapses or in growth cones. To understand how the cytoskeleton caters to these different demands, this review summarizes five decades of electron microscopic studies. It focuses on the organization of microtubules and neurofilaments in axon shafts in both vertebrate and invertebrate neurons, as well as the axon initial segments of vertebrate motor- and interneurons. Findings from these ultrastructural studies are being interpreted here on the basis of our contemporary molecular understanding. They strongly suggest that axon architecture in animals as diverse as arthropods and vertebrates is dependent on loosely cross-linked bundles of microtubules running all along axons, with only minor roles played by neurofilaments.

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“…The methodology of 4polar-STORM is also compatible with two-color localization and orientation measurements and thus can provide insights into the functional interplay between the nanometric organizations of interacting partners; the link between conformational changes in activated integrins and actin filament remodeling is such an example. At last, 4polar-STORM is compatible with 3D localization schemes, including astigmatism 61 or multiplane 62,63 strategies, and can therefore be adapted for exploring the full 3D organization of a large variety of biological structures 64 .…”

Section: Discussionmentioning

confidence: 99%

4polar-STORM polarized super-resolution imaging of actin filament organization in cells

Rimoli

Cav

Curcio

et al. 2021

Preprint

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Advances in single-molecule localization microscopy are providing unprecedented insights into the nanometer-scale organization of protein assemblies in cells and thus a powerful means for interrogating biological function. However, localization imaging alone does not contain information on protein conformation and orientation, which constitute additional key signatures of protein function. Here, we present a new microscopy method which combines for the first time Stochastic Optical Reconstruction Microscopy (STORM) super-resolution imaging with single molecule orientation and wobbling measurements using a four polarization-resolved image splitting scheme. This new method, called 4polar-STORM, allows us to determine both single molecule localization and orientation in 2D and to infer their 3D orientation, and is compatible with high labelling densities and thus ideally placed for the determination of the organization of dense protein assemblies in cells. We demonstrate the potential of this new method by studying the nanometer-scale organization of dense actin filament assemblies driving cell adhesion and motility, and reveal bimodal distributions of actin filament orientations in the lamellipodium, which were previously only observed in electron microscopy studies. 4polar-STORM is fully compatible with 3D localization schemes and amenable to live-cell observations, and thus promises to provide new functional readouts by enabling nanometer-scale studies of orientational dynamics in a cellular context.

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Three‐dimensional organization of the cytoskeleton: A cryo‐electron tomography perspective

Cited by 28 publications

References 184 publications

Considerations of Antibody Geometric Constraints on NK Cell Antibody Dependent Cellular Cytotoxicity

Considerations of Antibody Geometric Constraints on NK Cell Antibody Dependent Cellular Cytotoxicity

Cytoskeletal organization of axons in vertebrates and invertebrates

4polar-STORM polarized super-resolution imaging of actin filament organization in cells

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