Mammalia

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    Protocols in Current Issue
    Maternal Immune Activation with the Viral Mimetic Poly:IC in Pregnant Rats
    Authors:  Thaisa Meira Sandini, Quentin Greba, Brittney Rose Lins and John George Howland, date: 11/20/2020, view: 420, Q&A: 0

    Maternal immune activation (MIA) is increasingly well appreciated as an environmental risk factor for some psychiatric disorders. Administration of proinflammatory compounds such as the synthetic double-stranded RNA molecule

    ...
    Generating Three-dimensional Human Granulomas in vitro to Study Mycobacterium tuberculosis-host Interaction
    Authors:  Ainhoa Arbués, Michael Kammüller and Damien Portevin, date: 11/20/2020, view: 458, Q&A: 0
    Granulomas are organized multicellular structures that constitute the hallmark of an infection by the human pathogen Mycobacterium tuberculosis (Mtb). A better understanding of the complex host-Mtb interactions within the ...
    In vitro Time-lapse Imaging of Primary Cilium in Migrating Neuroblasts
    Neuronal migration is a critical step for the development of neuronal circuits in the brain. Immature new neurons (neuroblasts) generated in the postnatal ventricular-subventricular zone (V-SVZ) show a remarkable potential to migrate for a long ...
    Confocal Microscopy of Reovirus Transport in Living Dorsal Root Ganglion Neurons
    Authors:  Pavithra Aravamudhan, Krishnan Raghunathan and Terence S. Dermody, date: 11/20/2020, view: 381, Q&A: 0

    Neurotropic reoviruses repurpose host machinery to traffic over long distances in neuronal processes and access distal replication sites. Understanding mechanisms of neuronal transmission is facilitated by using simplified in vitro primary neuronal

    ...
    Visual-looming Shadow Task with in-vivo Calcium Activity Monitoring to Assess Defensive Behaviors in Mice
    There has been a clear movement in recent years towards the adoption of more naturalistic experimental regimes for the study of behavior and its underlying neural architecture. Here we provide a protocol that allows experimenters working with mice, ...
    Maternal Immune Activation with the Viral Mimetic Poly:IC in Pregnant Rats
    Authors:  Thaisa Meira Sandini, Quentin Greba, Brittney Rose Lins and John George Howland, date: 11/20/2020, view: 420, Q&A: 0
    [Abstract]

    Maternal immune activation (MIA) is increasingly well appreciated as an environmental risk factor for some psychiatric disorders. Administration of proinflammatory compounds such as the synthetic double-stranded RNA molecule polyinosinic-polycytidylic acid (polyI:C) to pregnant rodents results in the release of proinflammatory cytokines in the

    ...
    Generating Three-dimensional Human Granulomas in vitro to Study Mycobacterium tuberculosis-host Interaction
    Authors:  Ainhoa Arbués, Michael Kammüller and Damien Portevin, date: 11/20/2020, view: 458, Q&A: 0
    [Abstract] Granulomas are organized multicellular structures that constitute the hallmark of an infection by the human pathogen Mycobacterium tuberculosis (Mtb). A better understanding of the complex host-Mtb interactions within the granuloma’s environment may lead to new therapeutic or preventive tools to improve the control of the ...
    In vitro Time-lapse Imaging of Primary Cilium in Migrating Neuroblasts
    [Abstract] Neuronal migration is a critical step for the development of neuronal circuits in the brain. Immature new neurons (neuroblasts) generated in the postnatal ventricular-subventricular zone (V-SVZ) show a remarkable potential to migrate for a long distance at a high speed in the postnatal mammalian brain, and are thus a powerful model to analyze the ...
    Confocal Microscopy of Reovirus Transport in Living Dorsal Root Ganglion Neurons
    Authors:  Pavithra Aravamudhan, Krishnan Raghunathan and Terence S. Dermody, date: 11/20/2020, view: 381, Q&A: 0
    [Abstract]

    Neurotropic reoviruses repurpose host machinery to traffic over long distances in neuronal processes and access distal replication sites. Understanding mechanisms of neuronal transmission is facilitated by using simplified in vitro primary neuronal culture models. Advances in the design of compartmentalized microfluidic devices lend robustness to

    ...
    Visual-looming Shadow Task with in-vivo Calcium Activity Monitoring to Assess Defensive Behaviors in Mice
    [Abstract] There has been a clear movement in recent years towards the adoption of more naturalistic experimental regimes for the study of behavior and its underlying neural architecture. Here we provide a protocol that allows experimenters working with mice, to mimic a looming and advancing predatory threat from the sky. This approach is easy to implement ...
    Murine Acute Pneumonia Model of Pseudomonas aeruginosa Lung Infection
    Authors:  Xiaolei Pan and Weihui Wu, date: 11/05/2020, view: 402, Q&A: 0
    [Abstract] Animal infection models play significant roles in studying bacterial pathogenic mechanisms, host pathogen interaction as well as evaluating drug and vaccine efficacies. We have been utilizing an acute pneumonia model to study bacterial colonization in lungs and assess virulence to the host by determination of bacterial loads and survival assays, ...
    Super-resolution Imaging of the T cell Central Supramolecular Signaling Cluster Using Stimulated Emission Depletion Microscopy
    Authors:  Sin Lih Tan, Dominic Alibhai, Stephen J. Cross, Harry Thompson and Christoph Wülfing, date: 11/05/2020, view: 426, Q&A: 0
    [Abstract]

    Supramolecular signaling assemblies are of interest for their unique signaling properties. A µm scale signaling assembly, the central supramolecular signaling cluster (cSMAC), forms at the center interface of T cells activated by antigen presenting cells (APC). The adaptor protein linker for activation of T cells (LAT) is a key cSMAC component.

    ...
    Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels
    Authors:  Eric Parigoris, Daniel L. Dunkelmann and Unai Silvan, date: 11/05/2020, view: 778, Q&A: 0
    [Abstract] Giant unilamellar vesicles (GUVs) are a widely used model system for a range of applications including membrane biophysics, drug delivery, and the study of actin dynamics. While several protocols have been developed for their generation in recent years, the use of these techniques involving charged lipid types and buffers of physiological ionic ...
    Advances in Proximity Ligation in situ Hybridization (PLISH)
    Authors:  Monica Nagendran, Adam M Andruska, Pehr B Harbury and Tushar J Desai, date: 11/05/2020, view: 702, Q&A: 0
    [Abstract] Understanding tissues in the context of development, maintenance and disease requires determining the molecular profiles of individual cells within their native in vivo spatial context. We developed a Proximity Ligation in situ Hybridization technology (PLISH) that enables quantitative measurement of single cell gene expression ...
    In vitro Glutamylation Inhibition of Ubiquitin Modification and Phosphoribosyl-Ubiquitin Ligation Mediated by Legionella pneumophila Effectors
    Authors:  Alan G. Sulpizio, Marena E. Minelli and Yuxin Mao, date: 11/05/2020, view: 249, Q&A: 0
    [Abstract]

    Glutamylation is a posttranslational modification where the amino group of a free glutamate amino acid is conjugated to the carboxyl group of a glutamate side chain within a target protein. SidJ is a Legionella kinase-like protein that has recently been identified to perform protein polyglutamylation of the Legionella SdeA Phosphoribosyl-Ubiquitin

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