Antagonizing the fibroblast growth factor system in vitro and in vivo.
E. EREN KOCAK; C. A. TURNER; J. A. PEREZ; S. J. WATSON; H. AKIL
Society for Neuroscience. 2008.
The fibroblast growth factor (FGF) system has been implicated in affective disorders, including major depressive disorder in humans. We have previously found FGF2 to have antidepressant-like effects in animal models. However, there is a need for more specific antagonists of the FGF system. We designed and validated small interfering RNAs to FGF2 and FGF9 (Dharmacon). To this end, we transfected COS7 cells with a plasmid vector (psiCHECK-2, Promega) containing either FGF2 or FGF9 and co-transfected the cells with either FGF2 or FGF9 siRNA. We evaluated the tranfection efficiency, as well as the ability of siRNA to FGF2 or FGF9 to decrease protein levels by a dual-luciferase reporter assay. We subsequently determined the effects of FGF2 siRNA in a human gliomal cell line (U87MG) that endogenous expresses high levels of FGF2. We have identified sequences that are effective (>80% knockdown) and specific in vitro. We are currently evaluating the efficacy of antagonizing the FGF system in vivo by siRNA, as well as by pharmacological manipulation. We plan to assess the effects of antagonizing the FGF system on anxiety-like and depression-like behavior.