Imbalance in osteogenesis and adipogenesis of bone marrow stromal cells is a crucial pathological process of osteoporosis. P2 x 7-deficient mice were previously shown to exhibit an osteopenic phenotype and abnormal fat distribution, leading us to hypothesize that P2 x 7R activation was involved in the differentiation of BMSCs. Consequently, we investigated the effect of P2 x 7R activation on osteogenic and adipogenic differentiation of BMSCs in vitro, and established an ovariectomized (OVX) osteoporosis model to test P2 x 7R activation on adipocytes formation, trabecular and cortical bone parameters in vivo. Our results showed that activation of P2 x 7R by BzATP resulted in increase in the gene expression of osteoblastic markers, the activity of alkaline phosphatase and bone mineralization, and decrease in the gene expression of adipogenic markers and the number of adipocytes generated by BMSCs. MicroCT analysis showed that BzATP treatment ameliorated the micro-architecture of trabecular bones in OVX mice, while cortical bone parameters were unaffected. H&E staining analysis showed that was increase in the volume of trabecular bone and number of trabecular bone, and decrease in bone marrow adipocytes in BzATP-treated OVX mice compared with OVX mice. Also, activation of P2 x 7R transduced to ERK1/2 and JNK signaling pathways. This transduction was prevented by BBG, U0126, and SP600125. U0126 and SP600125 prevented BzATP-induced up-regulation of osteogenic-related genes expression and down-regulation of adipogenic-related genes expression. These data suggest that BzATP activates the differentiation of BMSCs into osteoblasts but not adipocytes by stimulating ERK1/2 and JNK signaling pathways in a P2 x 7R-dependent way. (C) 2015 Elsevier Inc. All rights reserved.