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Journal of Histochemistry and Cytochemistry, Vol. 51, 741-749, June 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Bioluminescent Detection of Endotoxin Effects on HIV-1 LTR-driven Transcription in Vivo

Fiona E. Yullc, Wei Hanc, E. Duco Jansenb, M. Brett Everharta, Ruxana T. Sadikota, John W. Christmana, and Timothy S. Blackwella
a Department of Medicine, Division of Allergy, Pulmonary and Critical Care Medicine, Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, and the Department of Veterans Affairs, Nashville, Tennessee
b Department of Biomedical Engineering, Vanderbilt University School of Medicine, and the Department of Veterans Affairs, Nashville, Tennessee
c Department of Cancer Biology, Vanderbilt University School of Medicine, and the Department of Veterans Affairs, Nashville, Tennessee

Correspondence to: Timothy S. Blackwell, Div. of Allergy, Pulmonary and Critical Care Medicine, Dept. of Medicine, Vanderbilt U. School of Medicine, T-1217 MCN, Nashville, TN 37232-2650. E-mail: timothy.blackwell@vanderbilt.edu

We investigated the effects of Gram-negative bacterial lipopolysaccharide (LPS) on luciferase expression in transgenic reporter mice in which luciferase expression is driven by the nuclear factor {kappa}B (NF-{kappa}B)-dependent portion of the human immunodeficiency virus-1 (HIV-1) long terminal repeat (HIV-1 LTR). Using these mice, we dissected the sources of luciferase activity at the organ level by (a) assessing luciferase activity in organ homogenates, (b) bioluminescence imaging in vivo, and (c) bioluminescence imaging of individual organs ex vivo. Luciferin dosage was a critical determinant of the magnitude of photon emission from these reporter mice. Photon emission increased at doses from 0.5–6 mg of luciferin given by intraperitoneal (IP) injection. The differential between basal and LPS-induced bioluminescence was maximal at 3–6 mg of luciferin. Luciferase expression was highly inducible in lungs, liver, spleen, and kidneys after a single IP injection of LPS, as assessed by luciferase activity measurements in organ homogenates. Luciferase activity was also induced in the forebrain by treatment with IP LPS. In contrast, aerosolized LPS produced a response localized to the lungs as assessed by both bioluminescence and ex vivo luciferase assay measurements. These studies demonstrate the utility of luciferase reporter mice for determining organ-specific gene expression in response to local and systemic stimuli. (J Histochem Cytochem 51:741–749, 2003)

Key Words: NF-{kappa}B, luciferase, transgenic, mouse


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