Open source software for quantification of cell migration, protrusions, and fluorescence intensities.
AuthorsBarry DJ, Durkin CH, Abella JV, Way M,
Journal
PubMed ID25847537
'Cell migration is frequently accompanied by changes in cell morphology (morphodynamics) on a range of spatial and temporal scales. Despite recent advances in imaging techniques, the application of unbiased computational image analysis methods for morphodynamic quantification is rare. For example, manual analysis using kymographs is still commonplace, often caused by ... More
A BRCA1-interacting lncRNA regulates homologous recombination.
AuthorsSharma V, Khurana S, Kubben N, Abdelmohsen K, Oberdoerffer P, Gorospe M, Misteli T,
Journal
PubMed ID26412854
Long non-coding RNAs (lncRNAs) are important players in diverse biological processes. Upon DNA damage, cells activate a complex signaling cascade referred to as the DNA damage response (DDR). Using a microarray screen, we identify here a novel lncRNA, DDSR1 (DNA damage-sensitive RNA1), which is induced upon DNA damage. DDSR1 induction ... More
Microtubule-dependent transport and dynamics of vimentin intermediate filaments.
We studied two aspects of vimentin intermediate filament dynamics-transport of filaments and subunit exchange. We observed transport of long filaments in the periphery of cells using live-cell structured illumination microscopy. We studied filament transport elsewhere in cells using a photoconvertible-vimentin probe and total internal reflection microscopy. We found that filaments ... More
Ebola Virus and Severe Acute Respiratory Syndrome Coronavirus Display Late Cell Entry Kinetics: Evidence that Transport to NPC1+ Endolysosomes Is a Rate-Defining Step.
AuthorsMingo RM, Simmons JA, Shoemaker CJ, Nelson EA, Schornberg KL, D'Souza RS, Casanova JE, White JM,
Journal
PubMed ID25552710
Ebola virus (EBOV) causes hemorrhagic fevers with high mortality rates. During cellular entry, the virus is internalized by macropinocytosis and trafficked through endosomes until fusion between the viral and an endosomal membrane is triggered, releasing the RNA genome into the cytoplasm. We found that while macropinocytotic uptake of filamentous EBOV ... More
3D imaging of Sox2 enhancer clusters in embryonic stem cells.
AuthorsLiu Z, Legant WR, Chen BC, Li L, Grimm JB, Lavis LD, Betzig E, Tjian R,
Journal
PubMed ID25537195
Combinatorial cis-regulatory networks encoded in animal genomes represent the foundational gene expression mechanism for directing cell-fate commitment and maintenance of cell identity by transcription factors (TFs). However, the 3D spatial organization of cis-elements and how such sub-nuclear structures influence TF activity remain poorly understood. Here, we combine lattice light-sheet imaging, ... More
The Nurr1 Activator 1,1-Bis(3'-Indolyl)-1-(p-Chlorophenyl)Methane Blocks Inflammatory Gene Expression in BV-2 Microglial Cells by Inhibiting Nuclear Factor ?B.
AuthorsDe Miranda BR, Popichak KA, Hammond SL, Jorgensen BA, Phillips AT, Safe S, Tjalkens RB,
Journal
PubMed ID25858541
NR4A family orphan nuclear receptors are an important class of transcription factors for development and homeostasis of dopaminergic neurons that also inhibit expression of inflammatory genes in glial cells. The identification of NR4A2 (Nurr1) as a suppressor of nuclear factor ?B (NF-?B)-related neuroinflammatory genes in microglia and astrocytes suggests that ... More
Axonal autophagosomes recruit dynein for retrograde transport through fusion with late endosomes.
AuthorsCheng XT, Zhou B, Lin MY, Cai Q, Sheng ZH,
Journal
PubMed ID25940348
Efficient degradation of autophagic vacuoles (AVs) via lysosomes is an important cellular homeostatic process. This is particularly challenging for neurons because mature acidic lysosomes are relatively enriched in the soma. Although dynein-driven retrograde transport of AVs was suggested, a fundamental question remains how autophagosomes generated at distal axons acquire dynein ... More
Posttranslational Modification of HOIP Blocks Toll-Like Receptor 4-Mediated Linear-Ubiquitin-Chain Formation.
AuthorsBowman J, Rodgers MA, Shi M, Amatya R, Hostager B, Iwai K, Gao SJ, Jung JU,
Journal
PubMed ID26578682
Linear ubiquitination is an atypical posttranslational modification catalyzed by the linear-ubiquitin-chain assembly complex (LUBAC), containing HOIP, HOIL-1L, and Sharpin. LUBAC facilitates NF-?B activation and inflammation upon receptor stimulation by ligating linear ubiquitin chains to critical signaling molecules. Indeed, linear-ubiquitination-dependent signaling is essential to prevent pyogenic bacterial infections that can lead ... More
Akt signaling dynamics in individual cells.
AuthorsGross SM, Rotwein P,
Journal
PubMed ID26040286
The protein kinase Akt is a key intracellular mediator of many biological processes, yet knowledge of Akt signaling dynamics is limited. Here we have constructed a fluorescent reporter molecule in a lentiviral delivery system to assess Akt kinase activity at the single cell level. The reporter, a fusion between a ... More
Visualization of Calcium Ion Loss from Rotavirus during Cell Entry.
AuthorsSalgado EN, Garcia Rodriguez B, Narayanaswamy N, Krishnan Y, Harrison SC
JournalJ Virol
PubMed ID30258012
'Bound calcium ions stabilize many nonenveloped virions. Loss of Ca'
Progressive Recruitment of Mesenchymal Progenitors Reveals a Time-Dependent Process of Cell Fate Acquisition in Mouse and Human Nephrogenesis.
AuthorsLindström NO, De Sena Brandine G, Tran T, Ransick A, Suh G, Guo J, Kim AD, Parvez RK, Ruffins SW, Rutledge EA, Thornton ME, Grubbs B, McMahon JA, Smith AD, McMahon AP
JournalDev Cell
PubMed ID29870722
'Mammalian nephrons arise from a limited nephron progenitor pool through a reiterative inductive process extending over days (mouse) or weeks (human) of kidney development. Here, we present evidence that human nephron patterning reflects a time-dependent process of recruitment of mesenchymal progenitors into an epithelial nephron precursor. Progressive recruitment predicted from ... More
AuthorsSteinmark IE, James AL, Chung PH, Morton PE, Parsons M, Dreiss CA, Lorenz CD, Yahioglu G, Suhling K
JournalPLoS One
PubMed ID30763333
'The only way to visually observe cellular viscosity, which can greatly influence biological reactions and has been linked to several human diseases, is through viscosity imaging. Imaging cellular viscosity has allowed the mapping of viscosity in cells, and the next frontier is targeted viscosity imaging of organelles and their microenvironments. ... More
A role for mitotic bookmarking of SOX2 in pluripotency and differentiation.
AuthorsDeluz C, Friman ET, Strebinger D, Benke A, Raccaud M, Callegari A, Leleu M, Manley S, Suter DM
JournalGenes Dev
PubMed ID27920086
'Mitotic bookmarking transcription factors remain bound to chromosomes during mitosis and were proposed to regulate phenotypic maintenance of stem and progenitor cells at the mitosis-to-G1 (M-G1) transition. However, mitotic bookmarking remains largely unexplored in most stem cell types, and its functional relevance for cell fate decisions remains unclear. Here we ... More
Unraveling growth factor signaling and cell cycle progression in individual fibroblasts.
AuthorsGross SM, Rotwein P,
JournalJ Biol Chem
PubMed ID27226630
Cultured cells require the actions of growth factors to enter the cell cycle, but how individual members of a population respond to the same stimulus remains unknown. Here we have employed continuous monitoring by live cell imaging in a dual-reporter cell model to investigate the regulation of short-term growth factor ... More
A PKC-MARCKS-PI3K regulatory module links Ca2+ and PIP3 signals at the leading edge of polarized macrophages.
AuthorsZiemba BP, Falke JJ
JournalPLoS One
PubMed ID29715315
The leukocyte chemosensory pathway detects attractant gradients and directs cell migration to sites of inflammation, infection, tissue damage, and carcinogenesis. Previous studies have revealed that local Ca2+ and PIP3 signals at the leading edge of polarized leukocytes play central roles in positive feedback loop essential to cell polarization and chemotaxis. ... More
Engineering a uniaxial substrate-stretching device for simultaneous electrophysiological measurements and imaging of strained peripheral neurons.
AuthorsBianchi F, George JH, Malboubi M, Jerusalem A, Thompson MS, Ye H
JournalMed Eng Phys
PubMed ID30878301
Peripheral nerves are continuously subjected to mechanical strain during everyday movements, but excessive stretch can lead to damage and neuronal cell functionality can also be impaired. To better understand cellular processes triggered by stretch, it is necessary to develop in vitro experimental methods that allow multiple concurrent measurements and replicate ... More
Membrane Mechanical Properties Regulate the Effect of Strain on Spontaneous Electrophysiology in Human iPSC-Derived Neurons.
AuthorsBianchi F, Pereno V, George JH, Thompson MS, Ye H
JournalNeuroscience
PubMed ID30817953
Peripheral nerves contain neuron fibers vital for movement and sensation and are subject to continuous elongation and compression during everyday movement. At supraphysiological strains conduction blocks occur, resulting in permanent or temporary loss of function. The mechanisms underpinning these alterations in electrophysiological activity remain unclear; however, there is evidence that ... More
Clinical trial in a dish: personalized stem cell derived cardiomyocyte assay compared to clinical trial results for two QT prolonging drugs.
Induced pluripotent stem cells (iPSCs) have shown promise in investigating donor-specific phenotypes and pathologies. iPSC-derived cardiomyocytes (iPSC-CMs) could potentially be utilized in personalized cardiotoxicity studies, assessing individual proarrhythmic risk. However, it is unclear how closely iPSC-CMs derived from healthy subjects can recapitulate a range of responses to drugs. It is ... More
Defects in COG-Mediated Golgi Trafficking Alter Endo-Lysosomal System in Human Cells.
The conserved oligomeric complex (COG) is a multi-subunit vesicle tethering complex that functions in retrograde trafficking at the Golgi. We have previously demonstrated that the formation of enlarged endo-lysosomal structures (EELSs) is one of the major glycosylation-independent phenotypes of cells depleted for individual COG complex subunits. Here, we characterize the ... More
A Mechanism of Resistance to Antibody-Targeted Immune Attack.
AuthorsAldeghaither DS, Zahavi DJ, Murray JC, Fertig EJ, Graham GT, Zhang YW, O'Connell A, Ma J, Jablonski SA, Weiner LM
JournalCancer Immunol Res
PubMed ID30563830
Targeted monoclonal antibody therapy is a promising therapeutic strategy for cancer, and antibody-dependent cell-mediated cytotoxicity (ADCC) represents a crucial mechanism underlying these approaches. The majority of patients have limited responses to monoclonal antibody therapy due to the development of resistance. Models of ADCC provide a system for uncovering immune-resistance mechanisms. ... More
Single-Cell Screening of Tamoxifen Abundance and Effect Using Mass Spectrometry and Raman-Spectroscopy.
AuthorsAli A, Abouleila Y, Shimizu Y, Hiyama E, Watanabe TM, Yanagida T, Germond A
JournalAnal Chem
PubMed ID30664349
Monitoring drug uptake, its metabolism, and response on the single-cell level is invaluable for sustaining drug discovery efforts. In this study, we show the possibility of accessing the information about the aforementioned processes at the single-cell level by monitoring the anticancer drug tamoxifen using live single-cell mass spectrometry (LSC-MS) and ... More