Figure 3: Strategy for BM-derived cell (BMDC)-specific BDNF deletion.
From: Haematopoietic cells produce BDNF and regulate appetite upon migration to the hypothalamus
(a) Schematic of the strategy for the creation of a congenital BDNF deletion specific for BMDCs (BM-BDNF−/−) by the interbreeding of mice of the Bdnftm3Jae/J and B6.129P2-Lyzstm1(cre)Ifo/J genotypes. Primers 1 and 2 were designed for the detection of genomic DNA. Primers 3 and 4 were designed for the detection of mRNA. (b) Immunostaining for BDNF and Iba1 in the PVN of BM-BDNF−/− and control LysM-cre mice (BM-BDNF+/+). The arrow indicates co-localization of staining for BDNF and Iba1 in the BM-BDNF+/+ cells, and the arrowhead indicates the lack of such co-localization in the BM-BDNF−/− cells. Scale bars, 10 μm. (c) Analysis of the co-localization of BDNF staining and GFP expression in GFP-TG/BM-BDNF−/− cells. GFP-TG/BM-BDNF−/− mice were generated by interbreeding of GFP-TG and BM-BDNF−/− mice. BDNF did not co-localize with GFP in GFP-TG/BM-BDNF-/- cells. Scale bars, 10 μm. (d) Genomic analysis of BM-BDNF−/−. The 1.3 kb BDNF gene is deleted in mononuclear cells and GFP-positive cells obtained from the PVN of GFP-TG/BM-BDNF−/− mice by LCM. (e) mRNA expression of BDNF. BDNF mRNA was not expressed in GFP-positive cells obtained from the PVN of GFP-TG/BM-BDNF−/− mice by LCM.
