NDLI: Dynamic imaging of pancreatic nuclear factor κB (NF-κB) activation in live mice using adeno-associated virus (AAV) infusion and bioluminescence.

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Dynamic imaging of pancreatic nuclear factor κB (NF-κB) activation in live mice using adeno-associated virus (AAV) infusion and bioluminescence.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Orabi, Abrahim I. Guo, Ping Prasadan, Krishna Jin, Shunqian Javed, Tanveer A. Sah, Swati Good, Misty L. Husain, Sohail Z. Xiao, Xiangwei Lemon, Kathryn L. Gittes, George K.
Description	Author Affiliation: Orabi AI ( From the Department of Pediatrics and.); Sah S ( From the Department of Pediatrics and.); Javed TA ( From the Department of Pediatrics and.); Lemon KL ( From the Department of Pediatrics and.); Good ML ( From the Department of Pediatrics and.); Guo P ( Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.); Xiao X ( Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.); Prasadan K ( Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.); Gittes GK ( Surgery, Children's Hospital of Pittsburgh of UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15224.); Jin S ( From the Department of Pediatrics and.); Husain SZ ( From the Department of Pediatrics and sohail.husain@chp.edu.)
Abstract	Nuclear factor κB (NF-κB) is an important signaling molecule that plays a critical role in the development of acute pancreatitis. Current methods for examining NF-κB activation involve infection of an adenoviral NF-κB-luciferase reporter into cell lines or electrophoretic mobility shift assay of lysate. The use of adeno-associated viruses (AAVs) has proven to be an effective method of transfecting whole organs in live animals. We examined whether intrapancreatic duct infusion of AAV containing an NF-κB-luciferase reporter (AAV-NF-κB-luciferase) can reliably measure pancreatic NF-κB activation. We confirmed the infectivity of the AAV-NF-κB-luciferase reporter in HEK293 cells using a traditional luciferase readout. Mice were infused with AAV-NF-κB-luciferase 5 weeks before induction of pancreatitis (caerulein, 50 µg/kg). Unlike transgenic mice that globally express NF-κB-luciferase, AAV-infused mice showed a 15-fold increase in pancreas-specific NF-κB bioluminescence following 12 h of caerulein compared with baseline luminescence (p < 0.05). The specificity of the NF-κB-luciferase signal to the pancreas was confirmed by isolating the pancreas and adjacent organs and observing a predominant bioluminescent signal in the pancreas compared with liver, spleen, and stomach. A complementary mouse model of post-ERCP-pancreatitis also induced pancreatic NF-κB signals. Taken together these data provide the first demonstration that NF-κB activation can be examined in a live, dynamic fashion during pancreatic inflammation. We believe this technique offers a valuable tool to study real-time activation of NF-κB in vivo.
ISSN	00219258
e-ISSN	1083351X
Journal	Journal of Biological Chemistry
Issue Number	18
Volume Number	290
Language	English
Publisher	American Society for Biochemistry and Molecular Biology (United States)
Publisher Date	2015-05-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Dependovirus Genetics Luminescent Measurements Molecular Imaging NF-kappa B Metabolism Pancreas Virology Animals Ceruletide Physiology HEK293 Cells Luciferases Mice Mice, Transgenic Organ Specificity Signal Transduction Research Support, N.I.H., Extramural Biochemistry Molecular Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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