Role of the hypoxia-inducible factor-1α in metabolic changes in the hypothalamus of obese mice

Project Description:

Astrocyte glucose metabolism is essential for the delivery of energy substrates to neurons and may regulate a variety of brain functions. The transcription factor hypoxia-inducible factor-1α (HIF-1α) regulates astrocytic glycolysis and lactate production (J Neurosci 32,9727, 2012). In obesity, low-grade inflammation occurs within the hypothalamus and astrocyte activity as well as glucose uptake is changed. Using in vitro and in vivo systems we will analyze the effect of dietary and inflammatory factors on HIF-1 signaling and target gene regulation in cultured astrocytes and within the hypothalamus of obese mice.

 

Experimental Methods:

• Primary cell cultures (astrocytes, microglia, brain endothelial cells), co-culture systems
• Mouse experiments including of rAAV vectors application and phenotyping (energy expenditure, body weight)
• DNA and RNA extraction, PCR, qPCR
• Western blotting, immunocytochemistry
• Chromatin Immunoprecipitation (ChIP), pyrosequencing of DNA methylation, reporter gene assays