Expression of Na(+)/Ca(2+) exchanger isoforms (NCX1 and NCX3) and plasma membrane Ca(2+) ATPase during osteoblast differentiation.
The ability to deliver calcium to the osteoid is critical to osteoblast function as a regulator of bone calcification. There are two known transmembrane proteins capable of translocating calcium out of the osteoblast, the Na(+)/Ca(2+) exchanger (NCX) and the plasma membrane Ca(2+)-ATPase (PMCA). In this study, we reveal the presence of the NCX3 isoform in primary osteoblasts and examine the expression of NCX1, NCX3, and PMCA1 during osteoblast differentiation. The predominant NCX isoform expressed by osteoblasts is NCX3. NCX1 also is expressed, but at low levels. Both NCX isoforms are expressed at nearly static levels throughout differentiation. In contrast, PMCA expression peaks at 8 days of culture, early in osteoblast differentiation, but declines thereafter. Immunocytochemical co-detection of NCX and PMCA reveal that NCX is positioned along surfaces of the osteoblast adjacent to osteoid, while PMCA is localized to plasma membrane sites distal to the osteoid. The expression pattern and spatial distribution of NCX support a role as a regulator of calcium efflux from osteoblasts required for calcification. The expression pattern and spatial distribution of PMCA makes its role in the mineralization process unlikely and suggests a role in calcium homeostasis following signaling events.
Stains
Joseph P
JP
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, 108 Althouse Lab, University Park, PA 16802, USA.
Weber
Janet A
JA
Gay
Carol V
CV
D000818:Animals; D002113:Calcification, Physiologic; D000252:Calcium-Transporting ATPases / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D027682:Cation Transport Proteins; D002454:Cell Differentiation; D002645:Chickens; D007150:Immunohistochemistry; D007700:Kinetics; D018613:Microscopy, Confocal; D008954:Models, Biological; D010006:Osteoblasts / Q000201:enzymology / Q000378:metabolism* / Q000502:physiology; D053499:Plasma Membrane Calcium-Transporting ATPases; D012333:RNA, Messenger / Q000096:biosynthesis; D019831:Sodium-Calcium Exchanger / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014158:Transcription, Genetic
D027682:Cation Transport Proteins; D012333:RNA, Messenger; D019831:Sodium-Calcium Exchanger; C110374:sodium-calcium exchanger 1; D053499:Plasma Membrane Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases