The Analysis of Osteoclasts Differentiation in Calvariae

颅骨破骨细胞分化分析

• 批准号: 09680732
• 负责人: MARUNOUCHI Tohru
• 金额: $ 1.98万
• 依托单位: Fujita Health University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680732/
• 关键词: Osteoclast; op; M-CSF; OPGL; TRAP-staining; Microglia; ERMP; op; M-CSF; ER-MP; TRAP; 頭蓋冠; サイトカイン

The analysis of osteoclasts differentiation in calvariae of op/op mouse. Recently, precursor cells of osteoclasts, which are induced from bone marrow stem cells by macrophage-colony stimulating factor (M-CSF), have been shown to differentiate into multinuclear osteoclasts at the bone tissue by the action of osteoprotegerin ligand (OPGL). However, precise biochemical reactions included in the differentiation process have to be elucidated. In the osteopetrotic, op/op mouse, the mutation of M-CSF gene prevents formation of macrophage and osteoclasts and, consequently, bone resorption. We have found that there are two sub-groups in the brain macrophages, microglia ; M-CSF dependent and M-CSF independent. The former is Mac-1, while the latter is ER-MP12 positive and can be detected in the op/op mouse. Since immature bone marrow cells share ER-MP antigen, it was conceivable that ER-MP12 positive precursor cells differentiate into osteoclasts at calvariae. TRAP-positive cells could not be found in the mixed glial culture prepared from op/op mouse. However, TRAP-negative calvariae of new born op/op mouse became TRAP-positive after culture for 13 days, suggesting the existence of the alternative pathway of osteoclast differentiation.

op/op小鼠颅骨破骨细胞分化的分析。最近，破骨细胞的前体细胞，其由骨髓干细胞通过巨噬细胞集落刺激因子（M-CSF）诱导，已经显示在骨组织中通过骨保护素配体（OPGL）的作用分化为多核破骨细胞。然而，精确的生化反应包括在分化过程中必须阐明。在骨硬化、OP/OP小鼠中，M-CSF基因突变阻止巨噬细胞和破骨细胞的形成，从而阻止骨吸收。我们发现脑巨噬细胞中有两个亚群，即小胶质细胞：M-CSF依赖性和M-CSF非依赖性。前者是Mac-1，而后者是ER-MP 12阳性，可以在op/op小鼠中检测到。由于未成熟的骨髓细胞共享ER-MP抗原，因此可以想象ER-MP 12阳性前体细胞在颅骨分化为破骨细胞。在op/op小鼠混合培养的胶质细胞中未发现TRAP阳性细胞。而新生op/op小鼠的颅骨在培养13 d后TRAP阴性变为阳性，提示破骨细胞分化的替代途径的存在。