쥐 골모세포 Na+-Ca2+ 교환이동체의 세포내 Ca2+ 농도 조절 기전

초록

Physiological activity of osteoblast cells including bone formation is known to be closely related to the increase in intracellular Ca2+ activity ([Ca2+]i) of osteoblast. Ca2+ is an important intracellular messenger in diverse cellular functions, and regulation of its level is mediated by the transmembrane Ca2+ movement via Ca2+ channels and Na+-Ca2+ exchange (NCX), and the intracellular Ca2+ movement through the intracellular stores. In this study, we investigated the role of NCX in regulation of intracellular Ca2+ activity in osteoblasts, employing antisense oligodeoxynucleotides technique. Osteoblasts were isolated from femur and tibia of neonatal rats, and cultured for 7-8 days. Antisense oligodeoxynucleotides (AS), 5‘-GTGAACATGACAAATGTCA-3‘ and sense (S), 5’-TGACATTTGTCATGTTCAC-3’ (bp3008-3026), were added into the culture media, on the 7th day. Cultured osteoblasts were loaded with Fura2-AM, and fluorescence images were monitored with a cooled CCD camera.