Journal: The Journal of Neuroscience
Article Title: FABP3 in the Anterior Cingulate Cortex Modulates the Methylation Status of the Glutamic Acid Decarboxylase 67 Promoter Region
Figure Lengend Snippet: Fabp3 gene ablation in the ACC caused upregulation of GAD67 expression and GABA synthesis. A, Quantitative analysis of the number of PV−, SOM−, and CR+ neurons in the ACC. PV, n = 8 sections, 4 mice; SOM, n = 12 sections, 6 mice; CR, n = 12 sections, 6 mice per group. B, Representative immunoblots (left) and quantitative densitometry analysis (right) probed with GABA-related antibodies. GAD67 and β-actin, n = 6 mice per group; GAD65, VGAT, and GABAAR, n = 5 mice per group; gephyrin, n = 5 and 6 WT and Fabp3 KO mice, respectively. C, Quantitative analysis of Gad67 mRNA expression in the ACC from WT and Fabp3 KO mice. n = 4 mice per group. D, Quantitative analysis of the GABA concentrations in ACC extracts from WT and Fabp3 KO mice by ELISA. n = 6 mice per group. E–G, There were no differences in GAD67 expression or GABA synthesis in the mPFC or AM of Fabp3 KO mice. E, Representative immunoblots (left) and quantitative densitometry analysis (right) probed with GAD67 antibody. n = 4 mice per group. F, Quantitative analysis of the Gad67 mRNA expression. n = 4 mice per group. G, Quantitative analysis of the GABA concentrations in the mPFC and AM extracts by ELISA. mPFC, n = 6 mice per group; AM, n = 4 and 6 WT and Fabp3 KO mice, respectively. H, I, Quantitative analysis of the DA and 5-HT concentrations in ACC extracts by ELISA. 5-HT, n = 6 mice per group; DA, n = 6 and 5 WT and Fabp3 KO mice, respectively. A–I, **p < 0.01 versus WT mice. J, Left, Representative immunoblots probed with FABP3 antibody. Right, Quantitative analysis of Gad67 mRNA in Neuro-2a cells. n = 4 per group. **p < 0.01 versus control. mock, Mock control; OE, Fabp3 overexpression; NC, negative control; KD, Fabp3 knockdown. Error bar indicates mean ± SEM.
Article Snippet: The primary antibodies included mouse monoclonal antibodies against FABP3 (1:200, Hycult Biotechnology HM2016, RRID: AB_533050 ); rabbit polyclonal antibodies against neurogranin (Ng, 1:2000, Millipore AB5620, RRID: AB_91937 ); rabbit polyclonal antibodies against PV (1:2000, Abcam ab11427, RRID: AB_298032 ); rat monoclonal antibodies against somatostatin (SOM, 1:50, Millipore MAB354, RRID: AB_2255365 ); and rabbit polyclonal antibodies against calretinin (CR, 1:2000, Swant CR7697, RRID: AB_2619710 ).
Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Over Expression, Negative Control