Solberg L B, Stang E, Brorson S-H, Andersson G, Reinholt F P
Department of Pathology, The Core Facility for Advanced Electron Microscopy, Oslo University Hospital, Rikshospitalet, P.O. Box 4950, Nydalen, 0424, Oslo, Norway,
Histochem Cell Biol. 2015 Feb;143(2):195-207. doi: 10.1007/s00418-014-1272-4. Epub 2014 Sep 9.
Tartrate-resistant acid phosphatase (TRAP) is well known as an osteoclast marker; however, a recent study from our group demonstrated enhanced number of TRAP + osteocytes as well as enhanced levels of TRAP located to intracellular vesicles in osteoblasts and osteocytes in experimental osteoporosis in rats. Such vesicles were especially abundant in osteoblasts and osteocytes in cancellous bone as well as close to bone surface and intracortical remodeling sites. To further investigate TRAP in osteoblasts and osteocytes, long bones from young, growing rats were examined. Immunofluorescence confocal microscopy displayed co-localization of TRAP with receptor activator of NF-KB ligand (RANKL) and osteoprotegerin (OPG) in hypertrophic chondrocytes and diaphyseal osteocytes with Pearson's correlation coefficient ≥0.8. Transmission electron microscopy showed co-localization of TRAP and RANKL in lysosomal-associated membrane protein 1 (LAMP1) + vesicles in osteoblasts and osteocytes supporting the results obtained by confocal microscopy. Recent in vitro data have demonstrated OPG as a traffic regulator for RANKL to LAMP1 + secretory lysosomes in osteoblasts and osteocytes, which seem to serve as temporary storage compartments for RANKL. Our in situ observations indicate that TRAP is located to RANKL-/OPG-positive secretory lysosomes in osteoblasts and osteocytes, which may have implications for osteocyte regulation of osteoclastogenesis.
抗酒石酸酸性磷酸酶(TRAP)是一种众所周知的破骨细胞标志物;然而,我们团队最近的一项研究表明,在实验性骨质疏松大鼠的成骨细胞和骨细胞中,TRAP+骨细胞数量增加,并且位于细胞内囊泡中的TRAP水平也有所提高。这种囊泡在松质骨的成骨细胞和骨细胞中尤其丰富,并且靠近骨表面和皮质内重塑部位。为了进一步研究成骨细胞和骨细胞中的TRAP,对幼年生长大鼠的长骨进行了检查。免疫荧光共聚焦显微镜显示,在肥大软骨细胞和骨干骨细胞中,TRAP与核因子κB受体活化因子配体(RANKL)和骨保护素(OPG)共定位,皮尔逊相关系数≥0.8。透射电子显微镜显示,在成骨细胞和骨细胞的溶酶体相关膜蛋白1(LAMP1)+囊泡中,TRAP和RANKL共定位,支持了共聚焦显微镜获得的结果。最近的体外数据表明,OPG是RANKL向成骨细胞和骨细胞中LAMP1+分泌性溶酶体转运的调节因子,这些溶酶体似乎是RANKL的临时储存区。我们的原位观察表明,TRAP位于成骨细胞和骨细胞中RANKL-/OPG阳性的分泌性溶酶体中,这可能对骨细胞对破骨细胞生成的调节有影响。