Establishment of stable reporter expression for in vivo imaging of nuclear factor-Κb activation in mouse liver

Abstract

The nuclear factor-ΚB (NF-ΚB) signaling pathway plays a critical role in a multitude of cellular processes. Activation of the NF-ΚB transcription factor family is essential for the initiation of inflammation, immunity, cell proliferation and apoptosis through a list of responsive genes. In hepatic tissue, activation of the NF-?B pathway has been implicated in a number of pathological conditions. Here we described a mouse model for noninvasive quantification of NF-ΚB activation in the hepatic tissues. Mice were subjected to hydrodynamic delivery with a mixture of pattB-NF-ΚB-Fluc reporter and φC31o integrase vector. Hepatic expression of φC31o integrase mediated chromosomal integration of the pattB-NF-ΚB-Fluc reporter, resulting in stable luciferase expression at 300 days post transfection. We applied noninvasive imaging and were able to detect NF-ΚB activation under acute liver injury and hepatitis conditions. During hepatectomy-induced liver regeneration, NF-ΚB activation was detected locally in the tissues at the surgery site. Treatment with Sorafenib suppressed NF-ΚB activation, accompanied with perturbation of liver regeneration. In conclusion, we established a method for stable transfection of the hepatic tissues and applied the transfected mice to longitudinal monitoring of NF-ΚB activity under pathological conditions. Further exploration of this methodology for establishment of other disease models and for evaluation of novel pharmaceuticals is likely to be fruitful. © Ivyspring International Publisher.