Journal: PLoS ONE
Article Title: Dendritic Branching of Olfactory Bulb Mitral and Tufted Cells: Regulation by TrkB
doi: 10.1371/journal.pone.0006729
Figure Lengend Snippet: A: OB neurons cultured from P1 thy1-YFP-G mice were doubly stained with anti-YFP antibody (A1; green in A2) and anti-TrkBex antibody (A3; red in A2) at 2 DIV. All YFP-positive cells coexpress TrkB. B: The p75NTR (red) is expressed by fibroblast-like cells and not by YFP-positive cells (green). Scale bars: 50 µm. C–E: Densities of DAPI (C) and YFP-positive cells (D) are measured after being cultured for 4 days with/without neurotrophins, and percentages of YFP-positive cells among DAPI are calculated and graphed after normalizing to control (E). BDNF, NT3, or NT4 were added to medium with final concentration of 1 or 10 ng/ml. When TrkBFc was applied, it was added to medium at a final concentration of 1 µg/ml with 10 ng/ml of neurotrophins. Application of BDNF or NT4 increases the proportion of YFP-positive cells in culture. Every condition was tested at least three times. Asterisks in E denote statistically significant differences from control condition analyzed with unpaired t test (**p<0.0001).
Article Snippet: After 3 hours of incubation at 37°C in a 5% CO 2 atmosphere, recombinant human neurotrophins (BDNF, NT3, or NT4, PeproTech Inc., Rocky Hill, NJ; 1 or 10 ng/ml) with/without recombinant human TrkBFc chimera proteins (R&D Systems; Minneapolis, MN; 1 μg/ml) were added to the medium.
Techniques: Cell Culture, Staining, Control, Concentration Assay