HISTOCHEMICAL SPECIFICITY OF STREPTOMYCES HYALURONIDASE AND CHONDROITINASE ABC
HISTOCHEMICAL SPECIFICITY OF STREPTOMYCES HYALURONIDASE AND CHONDROITINASE ABC
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DOI：
10.1007/bf01003135
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发表时间：
1978-01-01
期刊：
影响因子：
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通讯作者：
PINTAR, JE
中科院分区：
文献类型：
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作者：
DERBY, MA;PINTAR, JE
用降解特定类型糖胺聚糖的酶处理组织切片应提供一种在组织切片中定位糖胺聚糖的方法。通过利用鸡和鹌鹑胚胎中的内源标记糖胺聚糖检验了该技术的可行性。总糖胺聚糖中不到 8% 似乎在固定和脱水过程中非特异性损失。链霉菌透明质酸酶和软骨素酶 ABC 均降解了各自 90% 以上的底物，并且对其他糖胺聚糖的非特异性提取极少。通过将孵育缓冲液的 pH 值提高到 8.6，软骨素酶 ABC 对硫酸化糖胺聚糖的选择性大大提高。在此 pH 值下，软骨素酶 ABC 降解的透明质酸量可以忽略不计。使用链霉菌透明质酸酶和软骨素酶 ABC 证实，胚胎透明质酸在先前认为对硫酸化糖胺聚糖具有特异性的条件下与阿尔新蓝结合。这可能是由于与成人组织中的透明质酸相比，胚胎透明质酸的分子量增加。用缓冲液、pH 8.6 的软骨素酶 ABC 和链霉菌透明质酸酶处理相邻切片，然后用阿尔新蓝染色，可能提供了一种定位和定量组织切片中糖胺聚糖的方法。
Treatment of tissue sections with enzymes which degrade specific types of glycosaminoglycans should provide a means for localizing glycosaminoglycans in tissue sections. The feasibility of this technique was examined by utilizing endogenously labeled glycosaminoglycans in chick and quail embryos. Less than 8% of the total glycosaminoglycans appear to be lost non-specifically during fixation and dehydration. Both Streptomyces hyaluronidase and chondroitinase ABC degraded more than 90% of their respective substrates and demonstrated minimal non-specific extraction of other glycosaminoglycans. The selectivity of chondroitinase ABC for sulfated glycosaminoglycans was substantially increased by raising the pH of the incubation buffer to 8.6. At this pH, chondroitinase ABC degraded negligible amounts of hyaluronic acid. Use of both Streptomyces hyaluronidase and chondroitinase ABC confirmed that embryonic hyaluronic acid binds Alcian Blue under conditions that were previously believed specific for sulfated glycosaminoglycans. This may be due to the increased MW of embryonic hyaluronic acid compared to the hyaluronic acid in adult tissues. Treatment of adjacent sections with buffer, chondroitinase ABC at pH 8.6 and Streptomyces hyaluronidase and subsequent staining with Alcian Blue probably provides a method for localizing and quantitating glycosaminoglycans in tissue sections.