Cell Biology

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    Protocols in Current Issue
    Maximizing the Rod Outer Segment Yield in Retinas Extracted from Cattle Eyes
    Authors:  Isabella Panfoli, Daniela Calzia, Silvia Ravera, Paolo Bianchini and Alberto Diaspro, date: 07/20/2022, view: 565, Q&A: 0
    [Abstract]

    The retina is a thin neuronal multilayer responsible for the detection of visual information. The first step in visual transduction occurs in the photoreceptor outer segment. The studies on photoreception and visual biochemistry have often utilized rod outer segments (OS) or OS disks purified from mammalian eyes. Literature reports several OS and

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    Whole-mount Senescence-Associated Beta-Galactosidase (SA-β-GAL) Activity Detection Protocol for Adult Zebrafish
    Authors:  Marta Marzullo, Mounir El Maï and Miguel Godinho Ferreira, date: 07/05/2022, view: 1235, Q&A: 0
    [Abstract]

    Senescence-associated beta-galactosidase (SA-β-GAL) is an enzyme that accumulates in the lysosomes of senescent cells, where it hydrolyses β-galactosides. With p16, it represents a well-recognized biomarker used to assess senescence both in vivo and in cell culture. The use of a chromogenic substrate, such as

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    Experimental Models for Cold Exposure of Muscle in vitro and in vivo
    [Abstract]

    Work in cold environments may have a significant impact on occupational health. In these and similar situations, cold exposure localized to the extremities may reduce the temperature of underlying tissues. To investigate the molecular effects of cold exposure in muscle, and since adequate methods were missing, we established two experimental cold

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    Electroporation of Small Interfering RNAs into Tibialis Anterior Muscles of Mice
    Authors:  Anna Stephan, Flavia A. Graca, Liam C. Hunt and Fabio Demontis, date: 06/05/2022, view: 977, Q&A: 0
    [Abstract]

    Aging and wasting of skeletal muscle reduce organismal fitness. Regrettably, only limited interventions are currently available to address this unmet medical need. Many methods have been developed to study this condition, including the intramuscular electroporation of DNA plasmids. However, this technique requires surgery and high electrical

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    Plasma Membrane Wounding and Repair Assays for Eukaryotic Cells
    [Abstract]

    Damage to the plasma membrane and loss of membrane integrity are detrimental to eukaryotic cells. It is, therefore, essential that cells possess an efficient membrane repair system to survive. However, the different cellular and molecular mechanisms behind plasma membrane repair have not been fully elucidated. Here, we present three complementary

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    Simple Methods for Permanent or Transient Denervation in Mouse Sciatic Nerve Injury Models
    [Abstract]

    Our ability to move and breathe requires an efficient communication between nerve and muscle that mainly takes place at the neuromuscular junctions (NMJs), a highly specialized synapse that links the axon of a motor neuron to a muscle fiber. When NMJs or axons are disrupted, the control of muscle fiber contraction is lost and muscle are paralyzed.

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    Preparation of a Single Cell Suspension from the Murine Iridocorneal Angle
    Authors:  Benjamin R. Thomson and Susan E. Quaggin, date: 05/20/2022, view: 738, Q&A: 0
    [Abstract]

    Single cell RNA sequencing is a powerful tool that can be used to identify distinct cell types and transcriptomic differences within complex tissues. It has proven to be especially useful in tissues of the eye, where investigators have identified novel cell types within the retina, anterior chamber, and iridocorneal angle and explored

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    Protocol to Induce Follicular T Helper Cells, Germinal Centers, and Skin Lesions in Mouse Models for Skin Blistering Diseases
    [Abstract]

    Autoreactive T cells in autoantibody-mediated autoimmune diseases can be divided into two major subsets: (i) follicular T helper cells (Tfh) that provide T cell help in germinal centers (GC) and (ii) effector T (Teff) cells that immigrate into peripheral tissue sites such as the skin and mediate local inflammation. To study the sequence of events

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    Vestibular Organ Dissection and Whole-Mount Immunolabeling in Mouse
    Authors:  Timothy S. Balmer and Laurence O. Trussell, date: 05/20/2022, view: 664, Q&A: 0
    [Abstract]

    The vestibular sensory apparatus contained in the inner ear is a marvelous evolutionary adaptation for sensing movement in 3 dimensions and is essential for an animal’s sense of orientation in space, head movement, and balance. Damage to these systems through injury or disease can lead to vertigo, Meniere’s disease, and other disorders

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    Ex vivo Human Skin Infection with Herpes Simplex Virus 1
    Authors:  Nydia C. De La Cruz, Maureen Möckel, Lisa Wirtz and Dagmar Knebel-Mörsdorf, date: 05/05/2022, view: 630, Q&A: 0
    [Abstract]

    Although herpes simplex virus 1 (HSV-1) is a well-studied virus, how the virus invades its human host via skin and mucosa to reach its receptors and initiate infection remains an open question. For studies of HSV-1 infection in skin, mice have been used as animal models. Murine skin infection can be induced after injection or scratching of the

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