Acetylcholine is the predominant neurotransmitter in the neuromuscular junction, and a role in bone has been postulated. The expression of nicotinic receptors has been reported in osteoblasts, but the expression and function of muscarinic receptor in bone remain obscure. In this study, we investigated the expression and functional activities of muscarinic receptor subtypes in human osteoblast cell lines and animal and human bone tissue. The mRNA levels of muscarinic receptor subtypes were detected by reverse-transcription polymerase chain reaction. We found that muscarinic subtypes m1, m2, m3, m4, and m5 were expressed at different levels in human osteosarcoma HOS cells, rat femur, and human rib bone tissue; m1, m4, m5 were in cultured mouse femur bone cells and cultured mouse calvarial bone cells; m2, m3, m4 were in bovine bone. The mRNA of neuronal markers, light-, medium- and heavy-neurofilament, was not found in human bone tissues to exclude the possible contamination from neuronal tissue. Methacholine induced an elevation in cytosolic calcium concentration and proliferation in HOS cells. Both effects were blocked by atropine. We conclude that muscarinic receptor is present in bone tissue to evoke calcium signaling and modulate cell proliferation. Different muscarinic receptor subtypes are distributed in various parts of the animal skeletal system including the different species and bone portions. Bone remodeling involving osteoblast proliferation leads the possibilities that muscarinic receptor may play roles in bone remodeling.
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