acid-phosphatase and castanospermine

Osteoclasts are formed from hematopoietic precursors via cell-cell fusion. We have previously reported that mannose residues are expressed on the outer membranes of monocytes during osteoclast differentiation. In the present study, we have attempted to demonstrate the pattern of expression levels of terminal high-mannose type oligosaccharide and to show that the mannose receptor is expressed on osteoclast precursor cells. Osteoclasts were formed using three different systems, namely mouse bone marrow cell culture, co-culture of mouse spleen cells with stromal cells, and RAW264.7 cell cultures. During osteoclast differentiation, the expression of terminal high-mannose type oligosaccharide gradually increased and then peaked at the stage of fusion in all three systems. Expression of the mannose receptor gradually increased during osteoclast differentiation in bone marrow cells and the co-culture system. In contrast, that in RAW264.7 cells had already been detected in the absence of the soluble receptor activator of NF-kappaB ligand and did not change during osteoclast differentiation. To ascertain whether expression of high-mannose type oligosaccharide is involved in tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cell (MNC) formation, glycosidase inhibitors were used on RAW264.7 cell culture. Castanospermine, an inhibitor of glucosidase I, inhibited the TRAP-positive MNCs, and deoxymannojirimycin, an inhibitor of alpha-mannosidase I, increased the TRAP-positive MNC formation. These results indicate that the binding of terminal high-mannose and mannose receptor is important for the process of cellular fusion in osteoclast formation.

Topics: 1-Deoxynojirimycin; Acid Phosphatase; alpha-Glucosidases; alpha-Mannosidase; Animals; Biomarkers; Bone Marrow Cells; Cell Culture Techniques; Cell Differentiation; Cell Line; Cells, Cultured; Enzyme Inhibitors; Glycoside Hydrolase Inhibitors; Indolizines; Isoenzymes; Lectins, C-Type; Macrophages; Male; Mannose Receptor; Mannose-Binding Lectins; Mannosidases; Mice; Microscopy, Fluorescence; Oligosaccharides; Osteoclasts; Receptors, Cell Surface; Stem Cells; Tartrate-Resistant Acid Phosphatase