Acetylcholine affects osteocytic MLO-Y4 cells via acetylcholine receptors

• Acetylcholine receptors express in osteocytes and mediate cell functions.
• Acetylcholine receptors mediated the acetylcholine’s effects on osteocytes.
• We detected the expression of the acetylcholine receptors in MLO-Y4 cells.
• Acetylcholine affected the expression of osteocytic acetylcholine receptors.
• Acetylcholine regulated MLO-Y4 cell functions via the acetylcholine receptors.

The identification of the neuronal control of bone remodeling has become one of the many significant recent advances in bone biology. Cholinergic activity has recently been shown to favor bone mass accrual by complex cellular regulatory networks. Here, we identified the gene expression of the muscarinic and nicotinic acetylcholine receptors (m- and nAChRs) in mice tibia tissue and in osteocytic MLO-Y4 cells. Acetylcholine, which is a classical neurotransmitter and an osteo-neuromediator, not only influences the mRNA expression of the AChR subunits but also significantly induces the proliferation and viability of osteocytes. Moreover, acetylcholine treatment caused the reciprocal regulation of RANKL and OPG mRNA expression, which resulted in a significant increase in the mRNA ratio of RANKL:OPG in osteocytes via acetylcholine receptors. The expression of neuropeptide Y and reelin, which are two neurogenic markers, was also modulated by acetylcholine via m- and nAChRs in MLO-Y4 cells. These results indicated that osteocytic acetylcholine receptors might be a new valuable mediator for cell functions and even for bone remodeling.