Mammalia

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    Protocols in Current Issue
    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: 259, 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 ...
    Quantification of Prostaglandin E2 Concentration in Interstitial Fluid from the Hypothalamic Region of Free-moving Mice
    Authors:  Yoshinobu Nakamura, Takatoshi Sakaguchi, Ikumi Tamai and Takeo Nakanishi, date: 08/05/2019, view: 200, Q&A: 0
    [Abstract] Prostaglandin E2 (PGE2) is a well-established chemical mediator for the generation of the fever at the hypothalamus of the brain. PGE2 mediates fever generation via PGE receptor 3 (i.e., EP3) on neurons in the preoptic area. The role of PGE2 has been analyzed by measuring PGE2 ...
    Looking through Brains with Fast Passive CLARITY: Zebrafish, Rodents, Non-human Primates and Humans
    Authors:  Farzad Mortazavi, Alexander J. Stankiewicz and Irina V. Zhdanova, date: 08/05/2019, view: 223, Q&A: 0
    [Abstract] Recently developed CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ-hybridization-compatible Tis-sue-hYdrogel) technique renders the tissue transparent by removing lipids in the tissue, while preserving and stabilizing the cellular and subcellular structures. This provides effective penetration of diverse ...
    Precision Technique for Splenectomy Limits Mouse Stress Responses for Accurate and Realistic Measurements for Investigating Inflammation and Immunity
    Authors:  Shengwen Calvin Li, Anthony D. Rangel and Mustafa H. Kabeer, date: 08/05/2019, view: 221, Q&A: 0
    [Abstract] Splenectomy in an animal model requires a standardized technique utilizing best practice to avoid variability which can result in adverse impact to the animal resulting in flawed physiologic responses simply due to technique rather than to the studied variables. In the case of the spleen, often investigators are analyzing the animal immune or ...
    Isolation and Long-term Cultivation of Mouse Alveolar Macrophages
    [Abstract] Alveolar macrophages (AM) are tissue-resident macrophages that colonize the lung around birth and can self-maintain long-term in an adult organism without contribution of monocytes. AM are located in the pulmonary alveoli and can be harvested by washing the lungs using the method of bronchoalveolar lavage (BAL). Here, we compared different ...
    Isolation and Culture of Single Myofiber and Immunostaining of Satellite Cells from Adult C57BL/6J Mice
    Authors:  Shujie Chen, Hongrong Ding, Xiangping Yao and Liwei Xie, date: 07/20/2019, view: 524, Q&A: 0
    [Abstract] Myofiber isolation followed with ex vivo culture could recapitulate and visualize satellite cells (SCs) activation, proliferation, and differentiation. This approach could be taken to understand the physiology of satellite cells and the molecular mechanism of regulatory factors, in terms of the involvement of intrinsic factors over SCs ...
    Axon-seq for in Depth Analysis of the RNA Content of Neuronal Processes
    Authors:  Jik Nijssen, Julio Aguila and Eva Hedlund, date: 07/20/2019, view: 590, Q&A: 0
    [Abstract] Neuronal processes have an RNA composition that is distinct from the cell body. Therefore, to fully understand neuronal biology in health and disease we need to study both somas, dendrites and axons. Here we describe a detailed protocol of a newly refined method, Axon-seq, for RNA sequencing of axons (and dendrites) grown in isolation using single ...
    Isoelectric Focusing to Quantify Rhodopsin Phosphorylation in Mouse Retina
    Authors:  Sowmya Bekshe Lokappa, M. Carter Cornwall and Jeannie Chen, date: 07/20/2019, view: 1131, Q&A: 0
    [Abstract] Rhodopsin is a G-protein coupled receptor (GPCR) that mediates vision under dim light. Upon light exposure, rhodopsin is phosphorylated at multiple serine and threonine sites at its carboxyl-terminus by rhodopsin kinase (GRK1). This, in turn, reduces its ability to activate the visual G-protein transducin. Binding of light-activated, ...
    Protocol for Measuring Compulsive-like Feeding Behavior in Mice
    Authors:  Marta Fructuoso, Jose Espinosa-Carrasco, Ionas Erb, Cedric Notredame and Mara Dierssen, date: 07/20/2019, view: 393, Q&A: 0
    [Abstract] Obesity is an important health problem with a strong environmental component that is acquiring pandemic proportion. The high availability of caloric dense foods promotes overeating potentially causing obesity. Animal models are key to validate novel therapeutic strategies, but researchers must carefully select the appropriate model to draw the ...
    Lipid-exchange Rate Assay for Lipid Droplet Fusion in Live Cells
    Authors:  Jia Wang, Boon Tin Chua, Peng Li and Feng-Jung Chen, date: 07/20/2019, view: 470, Q&A: 0
    [Abstract] Lipid droplets (LDs) are central organelles in maintaining lipid homeostasis. Defective LD growth often results in the development of metabolic disorders. LD fusion and growth mediated by cell death–inducing DNA fragmentation factor alpha (DFFA)-like effector (CIDE) family proteins are crucial for various biological processes including unilocular ...



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