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The mechanism of cell locomotion of cultured osteoclasts in three-dimensions ; the observation by time-cinematography

培养破骨细胞三维运动机制；

基本信息

批准号：
04454449
负责人：
WAKITA Minoru
金额：
$ 4.29万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

Osteoclasts and macrophages of ddy strain mice were isolated, and then culturedwith or without dentine slices for 72 hours. After the incubation, tartrate-resistant acid phosphatase (TRACPase) and adic phosphateas (ACPase) activities were detected in them by the azo dye method. ACPasc-positive cells phagocytized indian inks, however, they showed neither TRACPase nor resosrptive activity. TRACPase-positive cells made lacunae on the dentine slices, and showed sthe ruffled border and clear zone. Therefore, ACPase-positive cells with phagocytosis were seemed to be macrophages, and TRACPase-positive cells were osteoclasts. These results suggested that TRACPase activity was specific for osteoclastic cells, not macrophages, in 72 hours culture of ddy mice.One multinucleated TRACPase-positive cell with comnplicated contours was observed on the dentine slice by light microscopy, and then this cell was serially sectioned by altermating semithin and ultrathin sections to observe its ultrastructure and three-dimensional sturcture. By transmission electron microscopy, this cell had the similar structure as those of osteoclasts, and possessed the clear zone-like structures, but showed no ruffled border. A three-dimensional reconstruction showed that this cell had very flat and complicated outlines, and extended its cell body to the back side of the dentine slice. These results suggested that this cell might be a locomoting osteoclast.

分离ddy品系小鼠的破骨细胞和巨噬细胞，分别用或不加牙本质切片培养72小时。孵育后，用偶氮染色法检测抗酒石酸酸性磷酸酶（TRACPase）和碱性磷酸酶（ACPase）活性。acpasc阳性细胞可吞噬印度墨水，但它们既没有显示出TRACPase活性，也没有显示出吸附活性。tracpase阳性细胞在牙本质切片上形成凹穴，边缘呈皱折状，带清晰。因此，具有吞噬作用的acpase阳性细胞似乎是巨噬细胞，而tracpase阳性细胞似乎是破骨细胞。这些结果表明，在培养72小时的小鼠中，TRACPase活性对破骨细胞特异性，而不是巨噬细胞。光镜下在牙本质切片上观察到1个轮廓复杂的多核tracpase阳性细胞，然后采用半薄和超薄交替连续切片，观察其超微结构和三维结构。透射电镜下，该细胞结构与破骨细胞相似，具有清晰的带状结构，但无皱褶边界。三维重建显示该细胞具有非常平坦复杂的轮廓，其细胞体延伸到牙本质切片的背面。这些结果提示该细胞可能是一种运动破骨细胞。