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We previously demonstrated, using osteoblastic MC3T3-E1 cells, that P2Y 2  purinergic receptors are involved in osteoblast mechanotransduction. In this study, our objective was to further investigate, using a knockout mouse model, the roles of P2Y 2  receptors in bone mechanobiology. We first examined bone structure with micro-CT and measured bone mechanical properties with three point bending experiments in both wild type mice and P2Y 2  knockout mice. We found that bones from P2Y 2  knockout mice have significantly decreased bone volume, bone thickness, bone stiffness and bone ultimate breaking force at 17 week old age. In order to elucidate the mechanisms by which P2Y 2  receptors contribute to bone biology, we examined differentiation and mineralization of bone marrow cells from wild type and P2Y 2  knockout mice. We found that P2Y 2  receptor deficiency reduces the differentiation and mineralization of bone marrow cells. Next, we compared the response of primary osteoblasts, from both wild type and P2Y 2  knockout mice, to ATP and mechanical stimulation (oscillatory fluid flow), and found that osteoblasts from wild type mice have a stronger response, in terms of ERK1/2 phosphorylation, to both ATP and fluid flow, relative to P2Y 2  knockout mice. However, we did not detect any difference in ATP release in response to fluid flow between wild type and P2Y 2  knock out osteoblasts. Our findings suggest that P2Y 2  receptors play important roles in bone marrow cell differentiation and mineralization as well as in bone cell mechanotransduction, leading to an osteopenic phenotype in P2Y 2  knockout mice.

The Roles of P2Y2 Purinergic Receptors in Osteoblasts and Mechanotransduction