Mammalia

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    Protocols in Current Issue
    Skin Transplantation and Lymphoid Organ Analysis in Mice
    [Abstract] Skin transplantation in mice is an important procedure to evaluate immune responses generated against heterologous grafts, especially given its highly immunogenic nature. In fact, skin is one of the most challenging organs in terms of allograft retention. In this protocol, we provide a detailed procedure for skin grafting using the tail skin as ...
    Visualizing Hypoxia in a Murine Model of Candida albicans Infection Using in vivo Biofluorencence
    Authors:  José Pedro Lopes and Constantin F. Urban, date: 08/05/2019, view: 1122, Q&A: 0
    [Abstract] Candida albicans is a leading human fungal pathogen that uses several metabolic adaptations to escape immune cells and causes systemic disease. Here, we describe a protocol for measuring one of these adaptations, the ability to thrive in hypoxic niches. Hypoxia was generated after successful subdermal infection with C. albicans ...
    Morphological Evaluation of Wound Healing Events in the Excisional Wound Healing Model in Rats
    [Abstract] Skin wound healing is a complex process involving different events such as blood coagulation, inflammation, new blood vessels formation, and extracellular matrix deposition. These events can be observed by using histology techniques. However, the lack of the standardization of such parameters impacts on the reproducibility of results. Here, we ...
    Activation of Fibroblast Contractility via Cell-Cell Interactions and Soluble Signals
    Authors:  Neha Pincha, Dyuti Saha, Tanay Bhatt, Ravindra K. Zirmire and Colin Jamora, date: 09/20/2018, view: 3869, Q&A: 0
    [Abstract] The collagen contraction assay is an in vitro, three-dimensional method to determine the factor(s) affecting the contractile behavior of activated cells such as fibroblasts in either physiological or pathological scenarios. The collagen lattices/hydrogels are seeded with fibroblasts to mimic the interactions between these cells and their ...
    Mouse Model of Immune Complex-mediated Vasculitis in Dorsal Skin and Assessment of the Neutrophil-mediated Tissue Damage
    [Abstract] Neutrophils are the most abundant leukocytes in the blood. In the recent decades, their crucial roles in host defense, immune regulation and tissue damage have been studied in a deeper dimension. In this protocol, we described a mouse model of immune complex-mediated vasculitis in the dorsal skin induced by Arthus reaction, and the subsequent ...
    A Novel Mouse Skin Graft Model of Vascular Tumors Driven by Akt1
    Authors:  Thuy L. Phung and Sriram Ayyaswamy, date: 07/05/2017, view: 6691, Q&A: 0
    [Abstract] To investigate whether endothelial Akt1 activation is sufficient to induce vascular tumor formation in the skin, we have developed a skin graft model in which a skin fragment from transgenic donor mice with inducible and endothelial cell-specific overexpression of activated Akt1 (myrAkt1) is grafted into the skin of wild type recipient mice. The ...
    Ear Inflammation and Whole-mount Ear Staining
    Authors:  Jia Tong Loh, Merry Gunawan and I-hsin Su, date: 10/20/2016, view: 6774, Q&A: 0
    [Abstract] The recruitment of circulating neutrophils from the bloodstream to the site of inflammation represents one of the earliest events during an innate immune response. During this response, neutrophils tether and roll along the vessel walls before transmigrating across the endothelium into the interstitial space to exert their functions. Here, we ...
    Skin TRITC Painting to Track Dendritic Cells Migrating to the Lymph Nodes
    Authors:  Jyh Liang Hor and Scott N. Mueller, date: 09/20/2016, view: 6010, Q&A: 0
    [Abstract] Our protocol describes a simple method that allows tracking of dendritic cells (DC) migration from the flank skin to draining lymph nodes (LN) using a red fluorescent dye tetramethylrhodamine-5-isothiocyanate (TRITC). TRITC is a photostable dye that readily labels cells including DC in the skin and can survive repeated exposure of two-photon laser ...
    Visualization of Intracellular Tyrosinase Activity in vitro
    Authors:  Riddhi Atul Jani, Sudeshna Nag and Subba Rao Gangi Setty, date: 04/20/2016, view: 5890, Q&A: 0
    [Abstract] Melanocytes produce the melanin pigments in melanosomes and these organelles protect the skin against harmful ultraviolet rays. Tyrosinase is the key cuproenzyme which initiates the pigment synthesis using its substrate amino acid tyrosine or L-DOPA (L-3, 4-dihydroxyphenylalanine). Moreover, the activity of tyrosinase directly correlates to the ...
    Estimation of Wound Tissue Neutrophil and Macrophage Accumulation by Measuring Myeloperoxidase (MPO) and N-Acetyl-β-D-glucosaminidase (NAG) Activities
    [Abstract] The inflammatory response is essential to the reestablishment of cutaneous homeostasis following injury. In this context, leukocytes arrive at the wound site and orchestrate essential events in the wound healing process. Therefore, the quantification of specific subsets of inflammatory cells in the wound tissue is of considerable interest. The ...



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