不同pH下的聚赖氨酸-金配合物是大鼠前牙本质和牙本质中糖胺聚糖和磷蛋白的鉴别检测探针。

The Histochemical Journal Pub Date : 1995-05-01
M Goldberg, S Lécolle
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摘要

在中性pH下,聚l -赖氨酸-金络合物广泛标记牙本质,而在牙本质中,金络合物的数量减少了一半。在标记前,在pH 6.8下对切片进行透明质酸酶预处理,抑制了牙本质前的大部分染色,不影响牙本质。pH为9的碱性磷酸酶预处理增强了牙本质前期的金配合物标记,去除了大部分牙本质中的金配合物标记。这证明在pH为7.2时,被染色的多阴离子包括位于牙本质前的糖胺聚糖和在牙本质中可见的磷蛋白的异质群体。在酸性pH值(2.9和1.1)下,被标记的金配合物数量减少,但牙本质前和牙本质标记之间的比例保持不变。透明质酸酶预处理去除或牢固地降低了牙本质前期和牙本质中的金络合物标记,而在标记前pH为9的碱性磷酸酶预处理没有引起任何变化。这证明了聚赖氨酸-金络合物染色对糖胺聚糖的特异性增加,在低pH值下染色在牙本质前和牙本质中。根据pH值对糖胺聚糖和磷酸蛋白进行区分染色,为研究这两种基质成分在牙本质矿化过程中所起的作用提供了有用的工具。
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Poly-L-lysine-gold complexes used at different pH are probes for differential detection of glycosaminoglycans and phosphoproteins in the predentine and dentine of rat incisor.

At neutral pH, poly-L-lysine-gold complexes labelled the predentine extensively, whereas in dentine the number of gold complexes was reduced by half. Hyaluronidase pretreatment of the section at pH 6.8, prior to labelling, suppressed most of the staining in predentine and did not affect dentine. In contrast, alkaline phosphatase pretreatment at pH 9 enhanced the gold complex labelling in predentine and removed most of the labelling in dentine. This proves that at pH 7.2, the polyanions which are stained include a heterogeneous population of glycosaminoglycans, located in predentine, and phosphoproteins, visualized in dentine. At acidic pH levels (2.9 and 1.1), the number of scored gold complexes decreased, but the ratio between predentine and dentine labelling remained constant. Hyaluronidase pretreatment removed or firmly reduced the gold complex labelling both in predentine and dentine, whereas alkaline phosphatase pretreatment of the sections at pH 9 prior to labelling did not induce any change. This argues in favour of an increased specificity of polylysine-gold complex staining for glycosaminoglycans, stained at low pH in both predentine and dentine. Differential staining of glycosaminoglycans and phosphoproteins according to the pH provides a useful tool for studying the role played respectively by the two matrix components in dentine mineralization.

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