Protocols in Current Issue
    Isolation of Pure Mitochondria from Rat Kidneys and Western Blot of Mitochondrial Respiratory Chain Complexes
    [Abstract] Cardiac, neuronal and renal tubular epithelial cells are the most metabolically active cells in the body. Their fate depends largely on their mitochondria as the primary energy generating system which participates in the control of apoptosis, cell cycle and metabolism. Thus, mitochondrial dysfunction is a hallmark of many chronic diseases ...
    Isolation and Transcriptomic Profiling of Single Myofibers from Mice
    [Abstract] Skeletal muscle is composed of different cells and myofiber types, with distinct metabolic and structural features. Generally, transcriptomic analysis of skeletal muscle is performed using whole muscle, resulting in average information as all cells composing the organ contribute to the expression value detected for each gene with the loss of ...
    Antisense Oligodeoxynucleotide Perfusion Blocks Gene Expression of Synaptic Plasticity-related Proteins without Inducing Compensation in Hippocampal Slices
    [Abstract] The elucidation of the molecular mechanisms of long-term synaptic plasticity has been hindered by both the compensation that can occur after chronic loss of the core plasticity molecules and by ex vivo conditions that may not reproduce in vivo plasticity. Here we describe a novel method to rapidly suppress gene expression by ...
    ATAC-seq on Sorted Adult Mouse Neurons
    Authors:  Erin A. Clark, Yasuyuki Shima and Sacha Nelson, date: 10/05/2019, view: 169, Q&A: 0
    [Abstract] Transcription regulation is a key aspect of cellular identity established during development and maintained into adulthood. Molecular and biochemical assays that probe the genome are critical tools in exploring mechanisms of transcription regulation and cell type identity. The mammalian brain is composed of a huge diversity of cell types with ...
    3D Organoid Formation from the Murine Salivary Gland Cell Line SIMS
    Authors:  Harleen K. Athwal and Isabelle M. A. Lombaert, date: 10/05/2019, view: 138, Q&A: 0
    [Abstract] Salivary glands consist of multiple phenotypically and functionally unique cell populations, such as the acinar, ductal, and myoepithelial cells that help produce, modify, and secrete saliva (Lombaert et al., 2011). Identification of mechanisms and factors that regulate these populations has been of key interest, as salivary gland-related ...
    An Operant Conditioning Task to Assess the Choice between Wheel Running and Palatable Food in Mice
    Authors:  Bastien Redon, Imane Hurel, Giovanni Marsicano and Francis Chaouloff, date: 10/05/2019, view: 203, Q&A: 0
    [Abstract] Wheel running, especially in the homecage, has been widely used to study the neurobiology of exercise because animal tends to use it voluntarily. However, as for each reward, its consumption (in the present case, running performance) does not specifically provide information on its incentive value, i.e., the extent to which animals are ...
    Imaging the Vasculature of Immunodeficient Mice Using Positron Emission Tomography/Computed Tomography (PET/CT) and 18F-fluorodeoxyglucose Labeled Human Erythrocytes
    Authors:  Shaowei Wang and Jung W. Choi, date: 10/05/2019, view: 101, Q&A: 0
    [Abstract] Nuclear blood pool imaging using radiolabeled red blood cells has been used in the clinical setting for the evaluation of a number of medical conditions including gastrointestinal hemorrhage, impaired cardiac contractility, and altered cerebrovascular blood flow. Nuclear blood pool imaging is typically performed using Technetium-99m-labeled (99m ...
    Simple Protocol for Distinguishing Drug-induced Effects on Spatial Memory Acquisition, Consolidation and Retrieval in Mice Using the Morris Water Maze
    Authors:  Ariel K. Frame, Asad Lone, Richard A. Harris and Robert C. Cumming, date: 09/20/2019, view: 188, Q&A: 0
    [Abstract] The Morris water maze (MWM) is one of the most commonly used tests for assessing spatial learning and memory in mice. While the MWM is highly amenable to testing the effects of memory modifying drugs, most studies do not consider the timing or duration of drug exposure when conducting the MWM assay; factors that can strongly influence the effect ...
    Quantitation of TLR4 Internalization in Response to LPS in Thioglycollate Elicited Peritoneal Mouse Macrophages by Flow Cytometry
    Authors:  Katharina Richard, Stefanie N. Vogel and Darren J. Perkins, date: 09/20/2019, view: 202, Q&A: 0
    [Abstract] Internalization of cell surface Toll-like Receptor 4 (TLR4) is a convenient read-out to measure LPS dependent activation of the TRIF adaptor pathway. We here provide a protocol to quantify the LPS dependent internalization of TLR4 using thioglycollate-elicited peritoneal macrophages by flow cytometry.
    Immunohistochemical Staining of CD8α in Diabetic Mouse Kidney
    Authors:  Ming-Zhi Zhang and Raymond C. Harris, date: 09/20/2019, view: 186, Q&A: 0
    [Abstract] Immune cell infiltration, particularly cytotoxic CD8α lymphocyte infiltration, plays an important role in development of diabetic nephropathy. Although CD8α infiltration can be evaluated by its production of cytokines, its localization in the kidney is of particular importance. The current protocol describes CD8α immunostaining using a Vectastain ...

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