Expression of the calcitonin receptor in bone marrow cell cultures and in bone: a specific marker of the differentiated osteoclast that is regulated by calcitonin.
Expression of the calcitonin receptor in bone marrow cell cultures and in bone: a specific marker of the differentiated osteoclast that is regulated by calcitonin.
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DOI：
10.1210/endo.136.10.7664679
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发表时间：
1995-10
期刊：
影响因子：
4.8
通讯作者：
Sun-Kyeong Lee;S. Goldring;J. Lorenzo
中科院分区：
文献类型：
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作者：
Sun-Kyeong Lee;S. Goldring;J. Lorenzo
我们研究了小鼠骨髓培养中破骨细胞（OC）从前体细胞分化的时间顺序。评估了OC表型的两种标志物，降钙素（CT）受体（CTR）和抗酒石酸酸性磷酸酶（TRAP）。将C57 BL/6小鼠的骨髓细胞在有或没有1，25-（OH）2维生素D3（10（-8）M）的情况下培养3、5、7和9天。在对照组中，每孔仅形成少量的骨肉瘤多核细胞（9 MNC）（第7天每孔80个）。TRAP的信使RNA（mRNA）在对照组和1，25-（OH）2D 3处理组中均通过逆转录-聚合酶链反应扩增检测到。然而，TRAP mRNA在MNCs中仅在第5、7和9天以及仅在1，25-（OH）2D 3处理的细胞中通过较不敏感的原位杂交检测到。在对照培养物中，CTR mRNA在第3天仅存在于非贴壁细胞中，并且在任何时间点均不存在于贴壁细胞（其中形成MNC）中。在1，25-（OH）2D 3处理的培养物中，在第3天的非贴壁细胞和第5天及之后的贴壁细胞中可检测到CTR mRNA。在第7天在贴壁细胞中观察到CTR mRNA的峰值水平（比第5天高15倍，比第9天高4倍）。CT（10（-7）M）处理7天培养物（其已被刺激以表达骨细胞表型）在24小时引起CTR mRNA表达的显著降低。CT处理对3 h的CTR mRNA表达或3或24 h的TRAP mRNA表达无影响。在新生小鼠颅盖培养物中，CTR mRNA的表达是组成性的，并且在CT处理48小时后显著降低。同样，在这些文化中的骨吸收抑制在24小时的CT治疗，但在48和72小时有逃脱CT对吸收的抑制作用。在骨髓培养物中，[125 I]-鲑鱼降钙素（sCT）结合的MNC阳性率大于98%，并且这种结合完全被过量冷sCT（10（-7）M）竞争消除。所有主要分离的破骨细胞从1至3天的小鼠长骨表现出[125 I]-sCT结合和TRAP活性，并强烈阳性CTR和TRAP mRNA的原位杂交。在骨髓培养物中形成的MNCs和分离的初级破骨细胞在骨切片上形成吸收陷窝。（400字处删节）
We studied the temporal sequence of osteoclast (OC) differentiation from precursor cells in murine marrow cultures. Two markers of the OC phenotype, calcitonin (CT) receptor (CTR) and tartrate resistant acid phosphatase (TRAP), were assessed. Marrow cells from C57BL/6 mice were cultured for 3, 5, 7, and 9 days with or without 1,25-(OH)2vitamin D3 (10(-8) M). In controls only small numbers of osteoclastic multinucleated cells 9MNCs) formed per well ( 80 per well on day 7). Messenger RNA (mRNA) for TRAP was detectable by reverse transcription-polymerase chain reaction amplification in both control and 1,25-(OH)2D3 treated groups at all times. However, TRAP mRNA was detectable in MNCs by the less sensitive in situ hybridization only on days 5, 7, and 9 and only in 1,25-(OH)2D3 treated cells. In control cultures, CTR mRNA was present on day 3 only in nonadherent cells and was not present in adherent cells (where MNCs formed) at any time point. In 1,25-(OH)2D3 treated cultures CTR mRNA was detectable in nonadherent cells on day 3 and in adherent cells on day 5 and thereafter. Peak levels of CTR mRNA were seen in adherent cells on day 7 (15-fold more than day 5 and 4-fold more than day 9). CT (10(-7) M) treatment of 7 day cultures, which had been stimulated to express the osteoclastic phenotype, caused a marked decrease in CTR mRNA expression at 24 h. There was no effect of CT treatment on CTR mRNA expression at 3 h or on TRAP mRNA expression at 3 or 24 h. In neonatal mouse calvaria cultures, CTR mRNA expression was constitutively present and was markedly decreased by 48 h of CT treatment. Similarly, bone resorption in these cultures was inhibited at 24 h by CT treatment, but at 48 and 72 h there was escape from the inhibitory effects of CT on resorption. In the marrow cultures, MNCs were greater than 98% positive for [125I]-salmon calcitonin (sCT) binding and this binding was completely competed away by excess cold sCT (10(-7) M). All primary isolated osteoclasts from 1- to 3-day-old mouse long bones exhibited [125I]-sCT binding and TRAP activity and were strongly positive for CTR and TRAP mRNA by in situ hybridization. Both MNCs that formed in bone marrow cultures and isolated primary osteoclasts formed resorption pits on bone slices.(ABSTRACT TRUNCATED AT 400 WORDS)