IMMUNOHISTOCHEMICAL DEMONSTRATION OF A 44-KD PHOSPHOPROTEIN IN DEVELOPING RAT BONES
IMMUNOHISTOCHEMICAL DEMONSTRATION OF A 44-KD PHOSPHOPROTEIN IN DEVELOPING RAT BONES
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DOI：
10.1177/35.7.3295029
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发表时间：
1987-07-01
影响因子：
3.2
通讯作者：
BUTLER, WT
中科院分区：
文献类型：
--
作者：
MARK, MP;PRINCE, CW;BUTLER, WT
本文用兔抗大鼠骨44-KD磷蛋白（44 K-BPP）的多克隆抗体，经亲和纯化后作为探针，研究了44 K-BPP在新生大鼠不同发育阶段骨中的分布。应用三种免疫染色程序，利用间接免疫荧光，抗生物素蛋白-生物素-过氧化物酶复合物，和抗生物素蛋白-金复合物与银增强。所有方法都给出了基本相同和/或互补的结果。在软骨和膜骨中检测到抗44 K BPP的抗原性。在后者中，它也表现在类骨质中。在下颌骨的编织骨中，在骨小梁之间但不与骨细胞外材料直接接触的成纤维细胞形细胞（前成骨细胞）中发现了抗44 K BPP抗体的免疫反应性。除了这些假定的骨祖细胞，成骨细胞以及骨细胞被强烈染色，细胞质染色与高尔基体。破骨细胞中偶尔检测到免疫反应性，但在这些细胞中，免疫染色要么弥漫性分布在细胞质中，要么仅存在于骨侵蚀部位。这些发现支持了44 K BPP是由成骨细胞产生的蛋白质并且主要定位于骨中的假设。此外，该蛋白似乎在骨形成细胞的组织发生早期表达。
Polyclonal antibodies against a 44-KD phosphoprotein (44K BPP) from rat bone were raised in rabbits, affinity-purified, and used as probes to study the protein''s distribution in various types of developing bones from newborn rats. Three immunostaining procedures were applied utilizing indirect immunofluorescence, avidin-biotin-peroxidase complex, and avidin-gold complex with silver enhancement. All methods gave essentially identical and/or complementary results. Antigenicity for anti-44K BPP was detected in endochondral and membranous bone. In the latter, it was also demonstrated in the osteoid. In the woven bone of lower jaw, immunoreactivity for anti-44K BPP antibodies was found in fibroblast-shaped cells (pre-osteoblasts) that were between the bone trabeculae but not in direct contact with bony extracellular material. In addition to these presumed osteoprogenitor cells, osteoblasts as well as osteocytes were strongly stained; the cytoplasmic staining was associated with the Golgi apparatus. Occasionally immunoreactivity was detected in osteoclasts, but in these cells immunostaining was either diffusely spread in the cytoplasm or present only at sites of bone erosion. These findings support the hypothesis that the 44K BPP is a protein made by osteoblasts and is localized predominantly in bone. Furthermore, the protein appears to be expressed early in histogenesis of the bone-forming cells.