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脱钙并在塑料中冷包埋后骨和软骨中的抗酒石酸酸性磷酸酶

Tartrate-resistant acid phosphatase in bone and cartilage following decalcification and cold-embedding in plastic.

作者信息

Cole A A, Walters L M

出版信息

J Histochem Cytochem. 1987 Feb;35(2):203-6. doi: 10.1177/35.2.3540104.

Abstract

Tartrate-resistant acid phosphatase (TRAP) has been proposed as a cytochemical marker for osteoclasts. We have developed an improved technique for the localization of TRAP in rat and mouse bone and cartilage. This procedure employs JB-4 plastic as the embedding medium, permits decalcification, and results in improved morphology compared with frozen sections. Peritoneal lavage cells were used to determine the appropriate isomer and concentration of tartrate necessary for inhibition of tartrate-sensitive acid phosphatase. After incubation in medium containing 50 mM L(+)-tartaric acid, osteoclasts and chondroclasts were heavily stained with reaction product. On the basis of their relative sensitivity to tartrate inhibition, three populations of mononuclear cells could also be distinguished. These three populations may represent: heavily stained osteoclast/chondroclast precursors; sparsely stained osteoblast-like cells lining the bone surface; and unstained cells of monocyte-macrophage lineage. Our results are consistent with the use of TRAP as a histochemical marker for study of the osteoclast.

摘要

抗酒石酸酸性磷酸酶(TRAP)已被提议作为破骨细胞的细胞化学标志物。我们开发了一种改进技术,用于在大鼠和小鼠的骨骼及软骨中定位TRAP。该方法采用JB - 4塑料作为包埋介质,允许脱钙,与冰冻切片相比,能改善形态。采用腹腔灌洗细胞来确定抑制酒石酸敏感酸性磷酸酶所需的合适酒石酸异构体和浓度。在含有50 mM L(+)-酒石酸的培养基中孵育后,破骨细胞和成软骨细胞被反应产物大量染色。根据它们对酒石酸抑制的相对敏感性,还可区分出三类单核细胞。这三类细胞可能分别代表:大量染色的破骨细胞/成软骨细胞前体;骨表面稀疏染色的成骨细胞样细胞;以及未染色的单核细胞 - 巨噬细胞系细胞。我们的结果与将TRAP用作研究破骨细胞的组织化学标志物是一致的。

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