Mammalia

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    Protocols in Current Issue
    Generation of Functional Mouse Hippocampal Neurons
    Authors:  Francesco Tomassoni-Ardori, Zhenyi Hong, Gianluca Fulgenzi and Lino Tessarollo, date: 08/05/2020, view: 519, Q&A: 0
    [Abstract] Primary culture of mouse hippocampal neurons is a very useful in vitro model for studying neuronal development, axonal and dendritic morphology, synaptic functions, and many other neuronal features. Here we describe a step-by-step process of generating primary neurons from mouse embryonic hippocampi (E17.5/E18.5). Hippocampal neurons ...
    In vivo Blood-brain Barrier Permeability Assays Using Clostridium perfringens Epsilon Toxin
    Authors:  Michael R. Mazzucco, Timothy Vartanian and Jennifer R. Linden, date: 08/05/2020, view: 267, Q&A: 0
    [Abstract] In order for the brain to function properly, a carefully orchestrated homeostasis must be maintained. To help regulate this delicate balance, the brain has developed a highly selective blood-brain barrier (BBB). Under normal conditions, the BBB excludes harmful blood-borne material from the brain parenchyma. However, numerous neuropathological ...
    Multiple Simultaneous Acute Stresses in Mice: Single or Repeated Induction
    [Abstract] Stress is crucial to the survival of an organism, but excessive stress can lead to psychological disorders including depression, anxiety, substance abuse, and suicidality. The prevailing notion is that chronic stress promotes adverse outcomes on brain and body health, whereas acute stressors are generally benign. Notably, acute events such mass ...
    Mechanical Tissue Compression and Whole-mount Imaging at Single Cell Resolution for Developing Murine Epididymal Tubules
    Author:  Tsuyoshi Hirashima, date: 08/05/2020, view: 460, Q&A: 0
    [Abstract] Cells inside the body are subjected to various mechanical stress, such as stretch or compression provided by surrounding cells, shear stresses by blood or lymph flows, and normal stresses by luminal liquids. Force loading to the biological tissues is a fundamental method to better understand cellular responses to such mechanical stimuli. There ...
    Development of the Deoxycorticosterone Acetate (DOCA)-salt Hypertensive Rat Model
    Authors:  Nayara Pestana-Oliveira, David B. Nahey, Tim Johnson and John P. Collister, date: 08/05/2020, view: 219, Q&A: 0
    [Abstract] According to the World Health Organization (WHO), nearly 1.13 billion people worldwide have hypertension, a major factor responsible for premature death globally. The inherent multifactorial nature of hypertension makes its study difficult since the chronic rise in blood pressure depends on the intricate connection between dietary, genetic ...
    Expression and Purification of Functionally Active Serotonin 5-HT2A Receptor in Insect Cells Using Low-titer Viral Stock
    [Abstract] The serotonin 5-HT2A receptor (5-HT2AR) is a member of the GPCR family that is important for various neurological functions and whose dysregulation causes many mental health disorders. Structural investigations of 5-HT2AR require the production of functionally active receptors expressed from eukaryotic cell ...
    Metabolomic and Lipidomic Analysis of Bone Marrow Derived Macrophages
    Authors:  Gretchen L. Seim, Steven V. John and Jing Fan, date: 07/20/2020, view: 335, Q&A: 0
    [Abstract] Macrophages are highly plastic immune cells that are capable of adopting a wide array of functional phenotypes in response to environmental stimuli. The changes in macrophage function are often supported and regulated by changes in cellular metabolism. Capturing a comprehensive picture of metabolism is vital for understanding the role of metabolic ...
    Preparation and Characterization of Ginger Lipid-derived Nanoparticles for Colon-targeted siRNA Delivery
    Authors:  Junsik Sung, Chunhua Yang, James F. Collins and Didier Merlin, date: 07/20/2020, view: 251, Q&A: 0
    [Abstract] Synthetic nanoparticle-based drug delivery system is widely known for its ability to increase the efficacy and specificity of loaded drugs, but it often suffers from relatively higher immunotoxicity and higher costs as compared to traditional drug formulations. Contrarily, plant-derived nanoparticles appear to be free from these limitations of ...
    Isolation, Culture, and Differentiation of Primary Myoblasts Derived from Muscle Satellite Cells
    Authors:  Kun Ho Kim, Jiamin Qiu and Shihuan Kuang, date: 07/20/2020, view: 302, Q&A: 0
    [Abstract] The skeletal muscle is key for body mobility and motor performance, but aging and diseases often lead to progressive loss of muscle mass due to wasting or degeneration of muscle cells. Muscle satellite cells (MuSCs) represent a population of tissue stem cells residing in the skeletal muscles and are responsible for homeostatic maintenance and ...
    Method for Prolonged Incubation of Brain Slices
    Authors:  Orsolya Kekesi and Yossi Buskila, date: 07/20/2020, view: 266, Q&A: 0
    [Abstract] Slices of neuronal tissue maintain a high degree of topographical and functional properties of neurons and glia and therefore are extensively used for measurements of neuronal activity at the molecular, cellular and network levels. However, the lifespan of slice preparations is narrow, averaging of 6-8 hours. Moreover, the average viability of ...



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