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分离的大鼠破骨细胞中存在丰富的降钙素受体。生化及放射自显影特征

Abundant calcitonin receptors in isolated rat osteoclasts. Biochemical and autoradiographic characterization.

作者信息

Nicholson G C, Moseley J M, Sexton P M, Mendelsohn F A, Martin T J

出版信息

J Clin Invest. 1986 Aug;78(2):355-60. doi: 10.1172/JCI112584.

Abstract

Calcitonin receptors have been characterized for the first time in isolated osteoclasts. These receptors have been demonstrated by autoradiographic and biochemical methods, and the cells have also been shown to respond to calcitonin with a dose-dependent increase in cyclic AMP. The receptors in rat osteoclasts are specific and of high affinity (dissociation constant, Kd, 1 to 6 X 10(-10) M), and are present in greater numbers than in any cell previously studied (greater than 10(6) per cell). Chemical cross-linking of 125I-labeled salmon calcitonin to osteoclasts using disuccinimidyl suberate resulted in identification of a receptor component with a relative molecular weight of 80,000-90,000. By counting grains in autoradiographic experiments, we found that greater than 80% of specifically bound radioactivity was associated with multinucleate osteoclasts and the remainder was associated with mononuclear cells that are not osteoblasts, but that may be osteoclast precursors.

摘要

降钙素受体首次在分离的破骨细胞中得到鉴定。这些受体已通过放射自显影和生化方法得以证实,并且这些细胞也已显示出对降钙素产生反应,伴随环磷酸腺苷呈剂量依赖性增加。大鼠破骨细胞中的受体具有特异性且亲和力高(解离常数Kd为1至6×10⁻¹⁰M),并且其数量比之前研究的任何细胞都多(每个细胞大于10⁶个)。使用辛二酸二琥珀酰亚胺酯将¹²⁵I标记的鲑鱼降钙素与破骨细胞进行化学交联,结果鉴定出一种相对分子质量为80,000 - 90,000的受体成分。通过对放射自显影实验中的银粒进行计数,我们发现超过80%的特异性结合放射性与多核破骨细胞相关,其余部分与非成骨细胞的单核细胞相关,但这些单核细胞可能是破骨细胞前体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bda0/423551/5947ab40a5e2/jcinvest00107-0036-a.jpg

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