Pharmacological and Genetic Accumulation of Hypoxia-Inducible Factor-1 Enhances Excitatory Synaptic Transmission in Hippocampal Neurons through the Production of Vascular Endothelial Growth Factor

Hypoxia-inducible factor-1 (HIF-1) is an important transcriptional factor in mammalian cells for coordination of adaptive responses to hypoxia. It consists of a regulatory subunit HIF-1 , which accumulates under hypoxic conditions, and a constitutively expressed subunit HIF-1 . In addition to the well characterized oxygen-dependent mode of action of HIF-1, recent work has shown that various growth factors and cytokines stimulate HIF-1 expression, thereby triggering transcription of numerous hypoxia-inducible genes by oxygenindependent mechanisms. In this study, we examined whether accumulation of HIF-1 induced by insulin-like growth factor-1 (IGF-1) has a regulatory role in excitatory synaptic transmission in hippocampal neuron cultures. Our results show that IGF-1 induced a timeand dose-dependent increase in HIF-1 expression that was blocked by pretreatment with selective IGF-1 receptor antagonist, transcriptional inhibitor, and translational inhibitors. In addition, pharmacological blockade of the phosphatidylinositol 3-kinase/Akt/mammalian target of the rapamycin signaling pathway, but not extracellular signal-regulated kinase, inhibited IGF-1-induced HIF-1 expression. More importantly, the increase in HIF-1 expression induced by IGF-1 was accompanied by increasing levels of vascular endothelial growth factor (VEGF) mRNA and protein, which enhanced excitatory synaptic transmission. In parallel, blockade of HIF-1 activity by echinomycin or lentiviral infection with dominant-negative mutant HIF-1 or short hairpin RNA targeting HIF-1 inhibited the increase in expression of VEGF and the enhancement of synaptic transmission induced by IGF-1. Conversely, transfection of constitutively active HIF-1 into neurons mimicked the effects of IGF-1 treatment. Together, these results suggest that HIF-1 accumulation can enhance excitatory synaptic transmission in hippocampal neurons by regulating production of VEGF.