A5056623924- CAR-T cell therapy research
- Immune Cell Function and Interaction
- Advanced Electron Microscopy Techniques and Applications
- Cellular transport and secretion
- Cancer Immunotherapy and Biomarkers
- Photosynthetic Processes and Mechanisms
- RNA and protein synthesis mechanisms
- Endoplasmic Reticulum Stress and Disease
- RNA Research and Splicing
- Lipid metabolism and biosynthesis
- Electron and X-Ray Spectroscopy Techniques
- Photoreceptor and optogenetics research
- Retinal Development and Disorders
- Advanced Fluorescence Microscopy Techniques
- Viral Infections and Immunology Research
- Lysosomal Storage Disorders Research
- Autoimmune and Inflammatory Disorders Research
- Diet, Metabolism, and Disease
- Phagocytosis and Immune Regulation
- Click Chemistry and Applications
- Liver Disease Diagnosis and Treatment
- Genomics and Chromatin Dynamics
- Lipid Membrane Structure and Behavior
- Neuroscience and Neuropharmacology Research
- Anodic Oxide Films and Nanostructures
Janelia Research Campus
2019-2023
Howard Hughes Medical Institute
2019-2022
The University of Melbourne
2022
University of California, Berkeley
2021
Within cells, the spatial compartmentalization of thousands distinct proteins serves a multitude diverse biochemical needs. Correlative super-resolution (SR) fluorescence and electron microscopy (EM) can elucidate protein relationships to global ultrastructure, but has suffered from tradeoffs structure preservation, retention, resolution, field view. We developed platform for three-dimensional cryogenic SR focused ion beam-milled block-face EM across entire vitreously frozen cells. The...
Lipid droplets (LDs) are neutral lipid storage organelles that transfer lipids to various including peroxisomes. Here, we show the hereditary spastic paraplegia protein M1 Spastin, a membrane-bound AAA ATPase found on LDs, coordinates fatty acid (FA) trafficking from LDs peroxisomes through two interrelated mechanisms. First, Spastin forms tethering complex with peroxisomal ABCD1 promote LD–peroxisome contact formation. Second, recruits membrane-shaping ESCRT-III proteins IST1 and CHMP1B via...
Chemical synapses between axons and dendrites mediate neuronal intercellular communication. Here, we describe a synapse primary cilia: the axo-ciliary synapse. Using enhanced focused ion beam-scanning electron microscopy on samples with optimally preserved ultrastructure, discovered brainstem serotonergic cilia of hippocampal CA1 pyramidal neurons. Functionally, these are enriched in ciliary-restricted serotonin receptor, 5-hydroxytryptamine receptor 6 (5-HTR6). cilia-targeted sensor, show...
Cytotoxic T lymphocytes (CTLs) and natural killer cells kill virus-infected tumor through the polarized release of perforin granzymes. Perforin is a pore-forming toxin that creates lesion in plasma membrane target cell which granzymes enter cytosol initiate apoptosis. Endosomal sorting complexes required for transport (ESCRT) proteins are involved repair small wounds. We found ESCRT were precisely recruited to sites CTL engagement immediately after release. Inhibition machinery...
Abstract mRNA translation is tightly regulated to preserve cellular homeostasis. Despite extensive biochemical, genetic, and structural studies, a detailed understanding of regulation lacking. Imaging methodologies able resolve the binding dynamics factors at single-cell single-mRNA resolution were necessary fully elucidate this paramount process. Here live-cell spectroscopy single-particle tracking combined interrogate endogenous initiation 5’cap. The diffusion (IFs) changed markedly upon...
Understanding cellular architecture is essential for understanding biology. Electron microscopy (EM) uniquely visualizes structures with nanometer resolution. However, traditional methods, such as thin-section EM or tomography, have limitations inasmuch they only visualize a single slice relatively small volume of the cell, respectively. Focused Ion Beam-Scanning Microscopy (FIB-SEM) demonstrated ability to image samples at 4-nm isotropic voxels rather limited imageable volume. Here, we...
Abstract Living cells function through the spatial compartmentalization of thousands distinct proteins serving a multitude diverse biochemical needs. Correlative super-resolution (SR) fluorescence and electron microscopy (EM) has emerged as pathway to directly view nanoscale protein relationships underlying global ultrastructure, but traditionally suffered from tradeoffs structure preservation, retention, resolution, field view. We developed platform for three-dimensional correlative...
Summary Chemical synapses between axons and dendrites mediate much of the brain’s intercellular communication. Here we describe a new kind synapse – axo-ciliary - primary cilia. By employing enhanced focused ion beam scanning electron microscopy on samples with optimally preserved ultrastructure, discovered serotonergic arising from brainstem, cilia hippocampal CA1 pyramidal neurons. Functionally, these are enriched in ciliary-restricted serotonin receptor, 5-hydroxytryptamine receptor 6...
Abstract Lipid droplets (LDs) are neutral lipid storage organelles that transfer lipids to various including peroxisomes. Here, we show the hereditary spastic paraplegia protein M1 Spastin, a membrane-bound AAA ATPase found on LDs, coordinates fatty acid (FA) trafficking from LDs peroxisomes through two inter-related mechanisms. First, Spastin forms tethering complex with peroxisomal ABCD1 promote LD-peroxisome contact formation. Second, recruits membrane-shaping ESCRT-III proteins IST1 and...
Abstract Despite extensive biochemical, genetic, and structural studies, a complete understanding of mRNA translation initiation is still lacking. Imaging methodologies able to resolve the binding dynamics factors at single-cell single-molecule resolution are necessary fully elucidate regulation this paramount process. We fused tags suitable for live imaging eIF4E, eIF4G1 4E-BP1 without affecting their function. combined Fluorescence Correlation Spectroscopy (FCS) Single-Particle Tracking...
Journal Article A Serotonergic Axon-Cilium Synapse Drives Nuclear Signaling to Maintain Chromatin Accessibility Get access Shu-Hsien Sheu, Sheu Janelia Research Campus, Ashburn, VA, USAHarvard Medical School, Boston, MA, USABoston Children’s Hospital, Department of Pathology, USAHHMI, Boston Cardiology, USA Correspondence to: sheus@janelia.hhmi.org, claphamd@janelia.hhmi.org Search for other works by this author on: Oxford Academic Google Scholar Srigokul Upadhyayula, Upadhyayula Advanced...
Elucidating the formation of molecular complexes in living cells is key for understanding cellular homeostasis, but has not been amenable to quantitative microscopy. Here we established a platform real-time quantification interactions monitor that results differential diffusion from their components. We have applied this one most dynamic processes: regulation mRNA translation. observe initiation factors (IFs) changes markedly upon association with mRNA. Quantifying binding dynamics revealed...