Expression of Extracellular Calcium (Ca2+o)-Sensing Receptor in Human Peripheral Blood Monocytes

• Published in: Biochemical and biophysical research communications 1998-05, Vol.246 (2), p.501-506
• Main Authors: Yamaguchi, Toru, Olozak, Ivona, Chattopadhyay, Naibedya, Butters, Robert R., Kifor, Olga, Scadden, David T., Brown, Edward M.
• Format: Article
• Language: English
• Subjects: Base Sequence; Calcium - blood; Cells, Cultured; DNA Primers - genetics; Extracellular Space - metabolism; Humans; Immunohistochemistry; Life Sciences (General); Monocytes - metabolism; Polymerase Chain Reaction; Receptors, Calcium-Sensing; Receptors, Cell Surface - blood; Receptors, Cell Surface - genetics; RNA, Messenger - blood; RNA, Messenger - genetics; Space life sciences
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1998.8648

The calcium-sensing receptor (CaR) is a G protein-coupled receptor playing key roles in extracellular calcium ion (Ca2+o) homeostasis in parathyroid gland and kidney. Macrophage-like mononuclear cells appear at sites of osteoclastic bone resorption during bone turnover and may play a role in the “reversal” phase of skeletal remodeling that follows osteoclastic resorption and precedes osteoblastic bone formation. Bone resorption produces substantial local increases in Ca2+othat could provide a signal for such mononuclear cells present locally within the bone marrow microenvironment. Indeed, previous studies by other investigators have shown that raising Ca2+oeitherin vivoorin vitrostimulated the release of interleukin-6 (IL-6) from human peripheral blood monocytes, suggesting that these cells express a Ca2+o-sensing mechanism. In these earlier studies, however, the use of reverse transcription-polymerase chain reaction (RT-PCR) failed to detect transcripts for the CaR previously cloned from parathyroid and kidney in peripheral blood monocytes. Since we recently found that non-specific esterase-positive, putative monocytes isolated from murine bone marrow express the CaR, we reevaluated the expression of this receptor in human peripheral blood monocytes. Immunocytochemistry, flow cytometry, and Western blot analysis, performed using a polyclonal antiserum specific for the CaR, detected CaR protein in human monocytes. In addition, the use of RT-PCR with CaR-specific primers, followed by nucleotide sequencing of the amplified products, identified CaR transcripts in the cells. Therefore, taken together, our data show that human peripheral blood monocytes possess both CaR protein and mRNA very similar if not identical to those expressed in parathyroid and kidney that could mediate the previously described, direct effects of Ca2+oon these cells. Furthermore, since mononuclear cells isolated from bone marrow also express the CaR, the latter might play some role in the “reversal” phase of bone remodeling, sensing local changes in Ca2+oresulting from osteoclastic bone resorption and secreting osteotropic cytokines or performing other Ca2+o-regulated functions that contribute to the control of bone turnover.