Distribution of Inositol 1,4,5-Trisphosphate Receptors in Rat Osteoclasts

Abstract

Inositol 1,4,5-trisphosphate (IP₃) receptors (IP₃Rs) are Ca²⁺ channels that localize to intracellular Ca²⁺ stores such as the endoplasmic reticulum (ER). Recently, IP₃Rs were found to participate in the formation of the cytoskeleton and cellular adhesions. In this study, we examined the cellular localization of type I, II, and III IP₃Rs to assess their role in cellular adhesion in rat osteoclasts. Rat bone marrow cells were cultured in α-MEM with 10% fetal bovine serum, M-CSF, RANKL, and 1,25(OH)₂D₃ for 1 week to promote osteoclast formation. Type I, II, and III IP₃R expression in the osteoclasts was then examined by RT-PCR. Double-staining was performed using antibodies against type I, II, and III IP₃Rs and DiOC₆, an ER marker, or TRITC-phalloidin, an actin filament marker. Expression of all three IP₃Rs was detected in the newly formed osteoclasts; however, the localization of the type I and II IP₃Rs was predominantly close to nuclear, and possibly colocalized with the ER, while the type III IP₃Rs were localized to the ER and podosomes, actin-rich adhesion structures in osteoclasts. These findings suggest that type III IP₃Rs are associated with osteoclast adhesion.