Brain-derived neurotrophic factor signaling is altered in the forebrain of Engrailed-2 knockout mice

Engrailed-2 (En2), a homeodomain transcription factor involved in regionalization and patterning of the midbrain and hindbrain regions has been associated to autism spectrum disorders (ASD). En2 knockout (En2-/-) mice show ASD-like features accompanied by a significant loss of GABAergic subpopulations in hippocampus and neocortex. Brain-derived neurotrophic factor (BDNF) is a crucial factor for the postnatal development of forebrain GABAergic neurons, and altered GABA signaling has been hypothesized to underlie the symptoms of ASD. Here we sought to determine whether interneuron loss in En2-/- forebrain might be related to altered expression of BDNF and its signaling receptors. We first evaluated the expression of different BDNF mRNA isoforms in the neocortex and hippocampus of wild-type (WT) and En2-/- mice. Quantitative RT-PCR showed a marked downregulation of several splicing variants of BDNF mRNA in the neocortex but not hippocampus of adult En2-/- mice, as compared to WT controls. Accordingly, levels of mature BDNF protein were lower in the neocortex but not hippocampus of En2-/- mice, as compared to WT. Increased levels of phosphorylated TrkB and decreased levels of p75 receptor were also detected in the neocortex of mutant mice. Accordingly, the expression of low density lipoprotein receptor (LDLR) and RhoA, two genes regulated via p75 was significantly altered in forebrain areas of mutant mice. These data indicate that BDNF signaling alterations might be involved in the anatomical changes observed in the En2-/- forebrain and suggest a pathogenic role of altered BDNF signaling in this mouse model of ASD.