title: Molecular imaging of the transcription factor NF-kappaB, a primary regulator of stress response.
abstract: A wide range of environmental stress and human disorders involves inappropriate regulation of NF-kappaB, including cancers and numerous inflammatory conditions. We have developed transgenic mice that express luciferase under the control of NF-kappaB, enabling real-time non-invasive imaging of NF-kappaB activity in intact animals. We show that, in the absence of stimulation, strong, intrinsic luminescence is evident in lymph nodes in the neck region, thymus, and Peyer's patches. Treating mice with stressors, such as TNF-alpha, IL-1alpha, or lipopolysaccharide (LPS) increases the luminescence in a tissue-specific manner, with the strongest activity observable in the skin, lungs, spleen, Peyer's patches, and the wall of the small intestine. Liver, kidney, heart, muscle, and adipose tissue exhibit less intense activities. Exposure of the skin to a low dose of UV-B radiation increases luminescence in the exposed areas. In ocular experiments, LPS- and TNF-alpha injected NF-kappaB-luciferase transgenic mice exhibit a 20-40-fold increase in lens NF-kappaB activity, similar to other LPS- and TNF-alpha-responsive organs. Peak NF-kappaB activity occurs 6h after injection of TNF-alpha and 12h after injection of LPS. Peak activities occur, respectively, 3 and 6h later than that in other tissues. Mice exposed to 360J/m(2) of UV-B exhibit a 16-fold increase in NF-kappaB activity 6h after exposure, characteristically similar to TNF-alpha-exposed mice. Thus, in NF-kappaB-luciferase transgenic mice, NF-kappaB activity also occurs in lens epithelial tissue and is activated when the intact mouse is exposed to classical stressors. Furthermore, as revealed by real-time non-invasive imaging, induction of chronic inflammation resembling rheumatoid arthritis produces strong NF-kappaB activity in the affected joints. Finally, we have used the model to demonstrate NF-kappaB regulation by manipulating the Vitamin A status in mice. NF-kappaB activity is elevated in mice fed a Vitamin A deficient (VAD) diet, and suppressed by surplus doses of retinoic acid (RA). We thus demonstrate the development and use of a versatile model for monitoring NF-kappaB activation both in tissue homogenates and in intact animals after the use of classical activators, during disease progression and after dietary intervention.
authors.affiliation: Department of Nutrition, Faculty of Medicine, University of Oslo, P.O. Box 1046 Blindern, N-0316 Oslo, Norway.
authors.forename: Harald
authors.initials: H
authors.lastname: Carlsen
authors.forename: George
authors.initials: G
authors.lastname: Alexander
authors.forename: Liv M I
authors.initials: LM
authors.lastname: Austenaa
authors.forename: Kanae
authors.initials: K
authors.lastname: Ebihara
authors.forename: Rune
authors.initials: R
authors.lastname: Blomhoff
mesh_terms: D000818:Animals; D003952:Diagnostic Imaging; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D007908:Lens, Crystalline / Q000737:chemistry; D008156:Luciferases / Q000032:analysis; D008163:Luminescent Measurements; D051379:Mice; D008822:Mice, Transgenic; D016328:NF-kappa B / Q000378:metabolism*; D014801:Vitamin A / Q000097:blood
chemical_list: D016328:NF-kappa B; D014801:Vitamin A; D008156:Luciferases