Molecular Mechanisms for Activation of the Agouti-Related Protein and Stimulation of Appetite
- Olha Ilnytska1,2,
- Adrian M. Stütz1,3,
- MieJung Park-York4,
- David A. York4,
- David M. Ribnicky5,
- Aamir Zuberi1,
- William T. Cefalu1 and
- George Argyropoulos1,6
- 1Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, Louisiana;
- 2Department of Biological Sciences, Rutgers University, Newark, New Jersey;
- 3Department of Genome Biology, European Molecular Biology Laboratory, Heidelberg, Germany;
- 4Center for Advanced Nutrition, Utah State University, Logan, Utah;
- 5Biotech Center, Cook College, Rutgers University, New Brunswick, New Jersey;
- 6Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania.
- Corresponding author: George Argyropoulos, .
OBJECTIVE The agouti-related protein (Agrp) is a powerful orexigenic peptide, but little is known about its transcriptional regulation. The objective of this study was to determine molecular mechanisms for the activation of hypothalamic Agrp and identify compounds that stimulate appetite.
RESEARCH DESIGN AND METHODS We used promoter analyses methods, hypothalamic cell culture and transfection, immunohistochemistry, luciferase-expressing transgenic mice, in vivo bioluminescence, anitisense RNA, mouse feeding studies, indirect calorimetry, real-time PCR, and Western blots.
RESULTS We found that the Krüppel-like factor 4 (Klf4) is a potent activator of Agrp by binding to a specific CACCC-box in its minimal promoter. We also found that an extract of tarragon, termed PMI-5011, activated hypothalamic Klf4 and Agrp. In vivo, PMI-5011 increased Agrp promoter activity in luciferase-expressing transgenic mice, increased hypothalamic Klf4 and Agrp expression, increased hypothalamic Orexin and melanin-concentrating hormone, increased food intake, reduced circulating insulin and leptin levels, attenuated energy expenditure, and enhanced body weight but only when using a high-fat diet.
CONCLUSIONS These data show that Klf4 augmented hypothalamic Agrp by binding to a specific CACCC-box onto its minimal promoter. In addition, the tarragon extract PMI-5011 activated Klf4 and orexigenic neuropeptides and reduced peripheral insulin and leptin levels leading to positive energy balance.
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- Received February 3, 2010.
- Accepted October 14, 2010.
- © 2011 by the American Diabetes Association.
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