Fig. 4: Ddah1 deficiency increases concentrations of ADMA to induce bone loss in vivo. | Nature Communications

Fig. 4: Ddah1 deficiency increases concentrations of ADMA to induce bone loss in vivo.

From: Mechanical force promotes dimethylarginine dimethylaminohydrolase 1-mediated hydrolysis of the metabolite asymmetric dimethylarginine to enhance bone formation

Fig. 4

a Serum and bone samples of 10-week old Ddah1+/+ and Ddah1−/− mice were analyzed by LC-MS assay. Ddah1+/+, n = 5. Ddah1−/−, n = 5. **p < 0.01. ***p < 0.005. Data are represented as mean values ± SD. b Serum and bone samples of 12-week-old Ddah1f/f and Ddah1prx1 mice were analyzed by an ADMA specific ELISA assay. Ddah1f/f, n = 5. Ddah1prx1, n = 5. *p < 0.05. **p < 0.01. Data are represented as mean values ± SD. c Immunofluorescence staining of eNOS (green) in bones of Ddah1+/+ and Ddah1−/− mice. Scale bar = 100 μm. d Immunofluorescence staining of NOS enzymes in bones of Ddah1f/f and Ddah1prx1 mice. Scale bar = 100 μm. e Mice were treated with saline solution or ADMA (12.5 mg/ml), and typical micro-CT images from tibiae are shown. Vehicle-treated group, n = 5, ADMA-treated group, n = 5. f Bone-microstructure analysis by micro-CT. The bone volume of total volume (BV/TV), trabecular bone separation (Tb.Sp), trabecular bone thickness (Tb.Th), trabecular bone number (Tb.N), and cortical bone thickness (Ct.Th) values are shown. *p < 0.05. Data are represented as mean values ± SD. g The serum ADMA concentrations were confirmed in ELISA assays. Vehicle-treated group, n = 5, ADMA-treated group, n = 5. **p < 0.01. Data are represented as mean values ± SD. h Serum P1NP concentrations were confirmed by performing ELISA assays. Vehicle-treated group, n = 4, ADMA-treated group, n = 4. **p < 0.01. Data are represented as mean values ± SD. i The BFR/BS ratio and MAR were analyzed by von Kossa staining and double-calcein staining. Scale bar = 100 μm. j Quantitative analysis of BFR/BS and MAR values. Vehicle-treated group, n = 5, ADMA-treated group, n = 5. *p < 0.05. Data are represented as mean values ± SD. k Bone biomechanics were analyzed by performing a three-point bending mechanics test. The bone stiffness and ultimate force are shown. Vehicle-treated group, n = 5, ADMA-treated group, n = 5. *p < 0.05. Data are represented as mean values ± SD. The data were analyzed by an unpaired two-tailed Student’s t test in two groups compared.

Back to article page