SIMULTANEOUS DEMONSTRATION OF BONE ALKALINE AND ACID-PHOSPHATASE ACTIVITIES IN PLASTIC-EMBEDDED SECTIONS AND DIFFERENTIAL INHIBITION OF THE ACTIVITIES
SIMULTANEOUS DEMONSTRATION OF BONE ALKALINE AND ACID-PHOSPHATASE ACTIVITIES IN PLASTIC-EMBEDDED SECTIONS AND DIFFERENTIAL INHIBITION OF THE ACTIVITIES
复制标题
DOI：
10.1007/bf00489547
复制
发表时间：
1987-01-01
期刊：
影响因子：
--
通讯作者：
HOWARD, GA
中科院分区：
文献类型：
--
作者：
LIU, C;SANGHVI, R;HOWARD, GA
骨碱性磷酸酶（AlP）和酸性磷酸酶（AcP）的活动，同时证明在从小鼠，大鼠和人类获得的组织切片。该方法涉及在乙醇中的组织固定，包埋在乙二醇甲基丙烯酸酯（GMA），和示范的AlP和AcP的活动，采用同时偶氮染料技术，取代萘酚磷酸盐作为底物。首先证明了AlP活性，然后是AcP活性。两种酶的活性在固定和/或储存在丙酮或70%乙醇中长达14天或储存在GMA中2个月的骨组织切片中得到证实。然而，在10%福尔马林、2%戊二醛或甲醛-钙中固定的骨组织切片中的AlP活性在3-7天后被显著抑制，并且在固定14天后不再检测到。此外，在70%乙醇或10%福尔马林中固定并随后在10%EDTA（pH 7）中脱矿2天的骨组织切片中，AlP活性降低，并且在随后在5%甲酸：20%柠檬酸钠（1：1，pH 4.2）中脱矿2天的骨组织切片中，该活性完全消除。甲基丙烯酸甲酯（MMA）包埋在66%以上的浓度完全抑制AlP活性。AcP活性，但是，只有部分抑制福尔马林，戊二醛，或福尔马林-钙后7或14天的固定或MMA包埋，是不受影响的脱矿质剂甲酸-柠檬酸盐2天。虽然固定在福尔马林中，随后在EDTA中脱矿的骨中保留了AcP活性，但在先前固定在70%乙醇中的骨上进行EDTA脱矿时，该活性完全被废除。这些结果表明，骨AlP和AcP的活动，可以同时证明在同一部分使用一个简单的组织制备技术，并保留在固定和/或储存在丙酮，70%乙醇，或GMA的组织的活动，但差异失活的其他固定剂研究和EDTA，甲酸-柠檬酸盐，MMA包埋。
Bone alkaline (AlP) and acid phosphatase (AcP) activities were simultaneously demonstrated in tissue sections obtained from mice, rats, and humans. The method involved tissue fixation in ethanol, embedding in glycol methacrylate (GMA), and demonstration of AlP and AcP activities employing a simultaneous coupling azo dye technique using substituted naphthol phosphate as a substrate. AlP activity was demonstrated first followed by AcP activity. Both enzyme activities were demonstrated in tissue sections from bones fixed and/or stored in acetone or 70% ethanol for up to 14 days or stored in GMA for 2 months. AlP activity in tissue sections from bones fixed in 10% formalin, 2% glutaraldehyde, or formal-calcium, however, was markedly inhibited after 3-7 days and was no longer detectable after 14 days of fixation. Moreover, AlP activity was diminished in tissue sections from bones fixed in 70% ethanol or 10% formalin and subsequently demineralized in 10% EDTA (pH 7) for 2 days, and the activity was completely abolished in tissue sections from bones subsequently demineralized in 5% formic acid: 20% sodium citrate (1:1, pH 4.2) for 2 days. Methyl methacrylate (MMA) embedding at concentrations above 66% completely inhibited AlP activity. AcP activity, however, was only partially inhibited by formalin, glutaraldehyde, or formal-calcium after 7 or 14 days of fixation or by MMA embedding and was unaffected by the demineralizing agent formic acid-citrate for 2 days. While AcP activity was preserved in bones fixed in formalin and subsequently demineralized in EDTA, the activity was completely abolished when EDTA demineralization was carried out on bones previously fixed in 70% ethanol. These results nidicate that bone AlP and AcP activities can be demonstrated simultaneously in the same section using a simple tissue preparation technique and that the activities are retained in tissues fixed and/or stored in acetone, 70% ethanol, or GMA, but are differentially inactivated by other fixatives studied and by EDTA, formic acid-citrate, and MMA embedding.