Protocols in Current Issue
    Bone-in-culture Array to Model Bone Metastasis in ex vivo Condition
    Authors:  Hai Wang and Xiang H.-F. Zhang, date: 01/20/2020, view: 35, Q&A: 0
    [Abstract] Bone is the most frequently affected organ by metastases of breast cancer and prostate cancer. Our knowledge on bone metastasis is extremely limited due to the lack of potent and efficient experimental models. We developed the “Bone-In-Culture Array (BICA)” platform to model the bone colonization of cancer cells in ex vivo cultures. The ...
    Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro
    Authors:  Nicholas D. Condon, Jennifer L. Stow and Adam A. Wall, date: 01/20/2020, view: 29, Q&A: 0
    [Abstract] Cell surface protrusions include F-actin rich, wave-like ruffles that are erected transiently in response to stimuli and during cell migration. Macrophages are innate immune cells that ruffle constitutively and more dramatically in cells activated by pathogens. Dorsal ruffles and their resulting macropinosomes are key sites for environmental ...
    A Simple and Efficient Method for Concomitant Isolation and Culture of Enriched Astroglial and Microglial Cells from the Rat Spinal Cord
    Authors:  Pooja Shree Mishra and Trichur R Raju, date: 01/20/2020, view: 117, Q&A: 0
    [Abstract] Investigations into glial biology have contributed substantially in understanding the physiology and pathology of the nervous system. However, intricacies of the neuron-glial and glial-glial interactions in vivo present significant challenges while delineating the individual cell-type contributions, thus making the in vitro ...
    Protocol for Measuring Free (Low-stress) Exploration in Rats
    Authors:  Wojciech Pisula and Klaudia Modlinska, date: 01/20/2020, view: 85, Q&A: 0
    [Abstract] Research on exploratory behavior plays a key part in behavioral science. Studying exploratory behavior of laboratory rodents may provide important data about many developmental and neurobiological processes occurring in animal ontogenesis. The proposed protocol for measuring the free (low-stress) exploration behavior in rats is straightforward, ...
    A Single Test to Study Social Behavior and Repetitive Self-grooming in Mice
    Authors:  Niraj V. Lawande, Ammar L. Ujjainwala and Catherine A. Christian, date: 01/20/2020, view: 79, Q&A: 0
    [Abstract] The ability to recognize and interact with members of the same species is essential for social communication. Investigating the neural substrates of social interest and recognition may offer insights into the behavioral differences present in disorders affecting social behavior. Assays used to study social interest in rodents include the 3-chamber ...
    Assessing Rough-and-tumble Play Behavior in Juvenile Rats
    Authors:  Jonathan W. VanRyzin, Ashley E. Marquardt and Margaret M. McCarthy, date: 01/05/2020, view: 292, Q&A: 0
    [Abstract] Play is a complex social behavior that is highly conserved across mammals. In most species, males engage in more frequent and vigorous play as juveniles than females, which reflects subtle yet impactful sex differences in brain circuitry and development. In this protocol, we describe a behavioral testing paradigm to assess social play in male and ...
    RNA Interactome Identification via RNA-BioID in Mouse Embryonic Fibroblasts
    Authors:  Joyita Mukherjee, Mirita Franz-Wachtel, Boris Maček and Ralf-Peter Jansen, date: 01/05/2020, view: 187, Q&A: 0
    [Abstract] Cytoplasmic localization of mRNAs is common to all organisms and serves the spatial expression of genes. Cis-acting RNA signals (mostly found in the mRNA’s 3'-UTR), called zipcodes recruit trans-acting RNA-binding proteins that facilitate the localization of the mRNA. UV-cross-linking or affinity purification has been applied to identify such ...
    Laser Capture Micro-dissection (LCM) of Neonatal Mouse Forebrain for RNA Isolation
    Authors:  Achira Roy, Mei Deng, Kimberly A. Aldinger, Ian A. GLass and Kathleen J. Millen, date: 01/05/2020, view: 243, Q&A: 0
    [Abstract] Precise and reproducible isolation of desired cell types or layers from heterogeneous tissues is crucial to analyze specific gene profiles and molecular interactions in vivo. Forebrain is the core site of higher functions, like cognition and memory consolidation. It is composed of heterogeneous and distinct cell types, interconnected to ...
    The Mouse Gambling Task: Assessing Individual Decision-making Strategies in Mice
    Authors:  Elsa Pittaras, Arnaud Rabat and Sylvie Granon, date: 01/05/2020, view: 165, Q&A: 0
    [Abstract] Decision-making is a complex cognitive process which consists of choosing one option among several alternatives. In humans, this process is featured in the Iowa gambling task (IGT), a decision-making task that mimics real life situations by reproducing uncertain conditions based on probabilistic rewards or penalties (see Background). Several ...
    Whole-cell and Perforated Patch-clamp Recordings from Acutely-isolated Murine Sino-atrial Node Cells
    Authors:  Qadeer Aziz, Muriel Nobles and Andrew Tinker, date: 01/05/2020, view: 123, Q&A: 0
    [Abstract] Cardiac pacemaker cells of the sino-atrial node are responsible for the initiation of the heart beat and express an array of ion channels. The patch-clamp technique is the gold standard method for investigating the function of ion channels expressed in electrically active cells. Conventional whole-cell and perforated patch-clamp techniques can be ...

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