NDLI: Intraductal injection of LPS as a mouse model of mastitis: signaling visualized via an NF-κB reporter transgenic.

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Intraductal injection of LPS as a mouse model of mastitis: signaling visualized via an NF-κB reporter transgenic.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Barham, Whitney Sherrill, Taylor Connelly, Linda Blackwell, Timothy S. Yull, Fiona E.
Description	Country affiliation: United States Author Affiliation: Barham W ( Cancer Biology Department, Vanderbilt University Medical Center, USA.)
Abstract	Animal models of human disease are necessary in order to rigorously study stages of disease progression and associated mechanisms, and ultimately, as pre-clinical models to test interventions. In these methods, we describe a technique in which lipopolysaccharide (LPS) is injected into the lactating mouse mammary gland via the nipple, effectively modeling mastitis, or inflammation, of the gland. This simulated infection results in increased nuclear factor kappa B (NF-κB) signaling, as visualized through bioluminescent imaging of an NF-κB luciferase reporter mouse. Our ultimate goal in developing these methods was to study the inflammation associated with mastitis in the lactating gland, which often includes redness, swelling, and immune cell infiltration. Therefore, we were keenly aware that incision or any type of wounding of the skin, the nipple, or the gland in order to introduce the LPS could not be utilized in our methods since the approach would likely confound the read-out of inflammation. We also desired a straight-forward method that did not require specially made hand-drawn pipettes or the use of micromanipulators to hold these specialized tools in place. Thus, we determined to use a commercially available insulin syringe and to inject the agent into the mammary duct of an intact nipple. This method was successful and allowed us to study the inflammation associated with LPS injection without any additional effects overlaid by the process of injection. In addition, this method also utilized an NF-κB luciferase reporter transgenic mouse and bioluminescent imaging technology to visually and quantitatively show increased NF-κB signaling within the LPS-injected gland. These methods are of interest to researchers of many disciplines who wish to model disease within the lactating mammary gland, as ultimately, the technique described here could be utilized for injection of a number of substances, and is not limited to only LPS.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/4030
Journal	Journal of Visualized Experiments
Issue Number	67
Language	English
Publisher	MyJove Corp.
Publisher Date	2012-09-04
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Disease Models, Animal Lipopolysaccharides Administration & Dosage Mastitis Chemically Induced Genetics Nf-kappa B Animals Genes, Reporter Metabolism Mice Mice, Transgenic Signal Transduction Research Support, N.i.h., Extramural Research Support, U.s. Gov't, Non-p.h.s. Video-audio Media
Content Type	Text
Resource Type	Article

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Intraductal Injection of LPS as a Mouse Model of Mastitis: Signaling Visualized via an NF-κB Reporter Transgenic

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Intraductal Injection of LPS as a Mouse Model of Mastitis: Signaling Visualized via an NF-κB Reporter Transgenic

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