Additional file 1:

Endogenous BDNF was expressed primarily along the mossy fiber pathways in cultured hippocampal slices, as observed in brain sections. Merged images of hippocampal slices stained (C, merged) with both Nissl staining of neurons (B) and BDNF immunohistochemistry (A). BDNF was highly expressed in the SL (arrow) and the dentate hilus, which are the normal mossy fiber pathways. GCL: granule cell layer, PCL: pyramidal cell layer, and DH: dentate hilus.

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Additional file 2:

Higher doses of K252a defasciculated the BDNF-overexpressing mossy fibers. We performed the same experiments as in the case of figure 3; however, we used higher doses [200 nM (A) and 300 nM (B)] of K252a. Mossy-fiber distribution was analyzed by determining the intensity of coexpressed Venus in the CA3c area (left) and the CA3a area (right). We observed a minor effect of the expressed BDNF at 200 nM (A); however, the effect was much lower than that observed in case of 100 nM K252a, suggesting that BDNF could overcome the effect of K252a at submaximal doses and that BDNF action on mossy fibers is dependent on Trk signaling. *P < 0.05 and **P < 0.01; Tukey's test after analysis of variance (n = 7–9 obtained from three independent experiments). SR: stratum radiatum, SP: stratum pyramidale and SO: stratum oriens.

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