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Cell biological studies on cell-cell, cell-matrix interaction in bone metabolism.

骨代谢中细胞-细胞、细胞-基质相互作用的细胞生物学研究。

基本信息

批准号：
02404071
负责人：
OZAWA Hidehiro
金额：
$ 16.7万
依托单位：
Niigata University.
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

We conducted 6 series of experiments on the correlation between osteoclasts and osteoblasts : 1) Changes in Ca concentration in osteoclasts was determined by fluorescence-labeling of Ca on single cells, and the effect of calcitonin on Ca movement using an Olympus OSP-3. Ca movement within osteoclasts was transiently enhanced by CT but rapidly decreased, after which the cells recovered their functional activities. The experiment was repeated on osteoclasts cultured on bone tissue, then the osteociasts were also submitted to a 2-dhnentional analysis using an Olympus CASARUS. Ca transport pathway within osteociasts and its inhibition by CT was evidenced. 2) Changes induced by CT on osteociast cytoskeleton was assessed with an Olympus IMT2-21CA3, by observing the changes in the localization of actin filametns and intermediate filaments occurring after addition of CT. 3) To clarify the differentiation of osteoclastic multinuclear giant cells (MNGCs), we cocultured osteoblasts and spleen cel … More ls in the presence of vitamin D3. The PMNGCs thus formed were TRACPasepositive and expressed CT receptors, which lead to consider them as osteociastic. Moreover, we found that a direct cell-to-cell contact with osteoblasts was required in the formation of osteoclastic MNGCs. 4) We also demonstrated by immunocytochemistry in vitro and in vivo that VDR was densely localized in the nucleus, especially in the heterochromatin, and that VDR was transported within the cytoplasm over and along microtubules to reach the nucleus. 5) We studied the interaction between differentiation/activation of osteoclasts and osteoblasts by lectin histochemistry and immunocytochemistry. Reaction of ConA, PNA, WGA, UEA-1 and MPA was intense on the membrane surface of osteoclast and osteoclasts where the two types of cells were in contact each other. The contact surface was also strongly heparan sulphate proteoglycan-positive. These two findings suggest an involvement of carbohydrate chains in the intercellular recognition between osteoclasts and osteoblasts, and to the local reservoir of growth factors and cytokines. 6) We attempted to clarify the inter-relationship between osteoclasts and osteocytes by cytochemistry, whereby we found that the latter cells enters in a cell-ceil interaction with the former : Osteocytes could promote bone resorption by this mechanism. Less

我们对破骨细胞和成骨细胞之间的相关性进行了6个系列的实验：1）通过荧光标记单个细胞上的Ca来测定破骨细胞中Ca浓度的变化，并且使用Olympus OSP-3测定降钙素对Ca运动的影响。破骨细胞内钙离子的运动被CT短暂增强，但很快下降，之后细胞恢复其功能活动。在骨组织上培养的破骨细胞上重复该实验，然后使用Olympus CASARUS对破骨细胞进行2-dnational分析。证明了成骨细胞内的钙转运途径及其被CT抑制。2)用OlympusIMT 2 - 21 CA 3型细胞骨架仪观察CT对成骨细胞骨架的影响，观察CT对成骨细胞骨架的影响。3)为了阐明成骨细胞多核巨细胞（MNGC）的分化，我们将成骨细胞和脾细胞共培养， ...更多信息是在维生素D3的存在下。由此形成的PMNGCs是TRACP酶阳性的，并表达CT受体，这导致认为它们是成骨细胞。此外，我们还发现成骨细胞与成骨细胞的直接细胞间接触是成骨细胞MNGCs形成所必需的。4)我们还通过体外和体内免疫细胞化学证明，VDR密集定位于细胞核，尤其是异染色质中，并且VDR在细胞质内通过并沿着微管运输到达细胞核。5)我们通过凝集素组织化学和免疫细胞化学研究破骨细胞和成骨细胞的分化/活化之间的相互作用。ConA、PNA、WGA、UEA-1和MPA在破骨细胞和破骨细胞接触处的膜表面有强烈的反应。接触表面也是强烈的硫酸乙酰肝素蛋白聚糖阳性。这两个发现表明碳水化合物链参与破骨细胞和成骨细胞之间的细胞间识别，并参与生长因子和细胞因子的局部储存。6)我们试图通过细胞化学来阐明破骨细胞和骨细胞之间的相互关系，由此我们发现后者细胞与前者进入细胞-细胞相互作用：骨细胞可以通过这种机制促进骨吸收。少