Figure 2

hBDNF-EGFP mRNA expression in tissues of three transgenic mouse lines. (A) RT-PCR analysis of hBDNF-EGFP mRNA expression in tissues of three transgenic BAC mouse lines – C3, E1, E4. mBDNF – mouse BDNF; hBDNF – human BDNF in human tissues; HPRT – reference gene hypoxanthine phosphoribosyltransferase. Cx – cortex; Hc – hippocampus; St – striatum; Th – thalamus; Mb – midbrain; PM – pons/medulla; Cb – cerebellum; Ty – thymus; He – heart; Lu – lung; Li – liver; Ki – kidney; SM – skeletal muscle; Sp – spleen; Te – testis. (B) Analysis of hBDNF-EGFP mRNA expression levels in C3 mouse brain by RNase protection assay. hBDNF-EGFP probe was used to determine both transgenic and endogenous BDNF mRNA levels as protein coding sequences of mouse and human BDNF share a high degree of similarity. P – probe without RNase; tRNA – yeast tRNA; HC – hippocampus; CX – cortex. On the right, black boxes denote vector-derived sequences, white boxes BDNF and gray boxes EGFP sequences. (C) Expression of alternative hBDNF-EGFP transcripts in C3 mouse hippocampus (HC), analyzed by RT-PCR. PCR primers used were specific for human BDNF transcripts as shown by control reactions with human (hHC) and mouse (mHC) hippocampal cDNA. eIX – transcript containing 5'-extended exon IX.