IMMUNOCHEMICAL AND IMMUNOHISTOCHEMICAL STUDIES, USING ANTISERA AGAINST PORCINE 25KDA AMELOGENIN, 89KDA ENAMELIN AND THE 13-17KDA NONAMELOGENINS, ON IMMATURE ENAMEL OF THE PIG AND RAT
IMMUNOCHEMICAL AND IMMUNOHISTOCHEMICAL STUDIES, USING ANTISERA AGAINST PORCINE 25KDA AMELOGENIN, 89KDA ENAMELIN AND THE 13-17KDA NONAMELOGENINS, ON IMMATURE ENAMEL OF THE PIG AND RAT
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DOI：
10.1007/bf00315983
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发表时间：
1991-01-01
期刊：
影响因子：
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通讯作者：
KOBAYASHI, S
中科院分区：
文献类型：
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作者：
UCHIDA, T;TANABE, T;KOBAYASHI, S
从新形成的未成熟猪牙釉质层中提取牙釉质蛋白，并纯化25 kDa牙釉蛋白、89 kDa牙釉蛋白和13-17 kDa非牙釉蛋白。 针对这些蛋白质产生了特异性抗血清。 通过用 20 kDa 牙釉蛋白（含有抗原的残基 1-148）吸收抗 25 kDa 牙釉蛋白血清来产生特异于 25 kDa 牙釉蛋白 C 端区域（残基 149-173）的抗体。 提取的猪牙釉质蛋白的免疫电转移印迹表明，抗25 kDa牙釉蛋白血清可识别25 kDa和其他低分子量和高分子量牙釉蛋白。 C末端特异性抗25kDa牙釉蛋白血清仅与分子量超过23kDa的牙釉蛋白反应。 抗 89 kDa 牙釉蛋白血清识别 89 kDa 牙釉蛋白和较低分子量蛋白质，但既不识别牙釉蛋白，也不识别 13-17 kDa 非牙釉蛋白。 针对 13-17 kDa nonamelogenins 的抗血清显示出与 89 kDa enamelin 没有交叉反应性，但识别出更高分子量的 nonamelogenins。 在猪牙胚的免疫组织化学制剂中，未成熟牙釉质上的 25 kDa 牙釉质样免疫反应性从牙釉质表面到中层呈梯度下降。 在内层中，免疫反应集中在棱柱鞘上。 C 端特异性 25 kDa 牙釉蛋白样免疫反应性在未成熟牙釉质的外层强烈，并向中层急剧下降。 棱柱鞘被 13-17 kDa nonamelogenins 的抗血清强烈染色。 牙釉质棱柱上的 89 kDa 类牙釉质免疫反应性在外层强烈，向中层减弱。 棱镜鞘的抗 89 kDa 牙釉质血清的染色很弱。 在未成熟的大鼠门牙釉质中，C 端特异性 25 kDa 釉原蛋白抗血清表现出与猪未成熟牙釉质相似的染色模式。 尤其是在 Tomes 突附近的层中，发现了不同的 13-17 kDa nonamelogenin 样和 89 kDa enamelin 样免疫反应性。 我们得出的结论是，一些牙釉质蛋白在从大鼠和猪的分泌性成釉细胞分泌后不久就会被降解。 与 13-17 kDa nonamelogenins 抗血清反应的特定牙釉质蛋白似乎与未成熟猪牙釉质中棱柱鞘的形成有关，但与大鼠门牙牙釉质中无关。
Enamel proteins were extracted from the newly formed layer of immature porcine enamel, and the 25 kDa amelogenin, 89 kDa enamelin and 13-17 kDa nonamelogenins were purified. Specific antisera were raised against these proteins. Antibodies specific to the C-terminal region (residues 149-173) of the 25 kDa amelogenin were generated by absorption of the anti-25 kDa amelogenin serum with 20 kDa amelogenin, which contains residues 1-148 of the antigen. Immunoelectro-transfer blotting of the extracted porcine enamel proteins showed that the anti-25 kDa amelogenin serum recognized the 25 kDa and other low and high molecular weight amelogenins. The C-terminal specific anti-25 kDa amelogenin serum reacted only with amelogenins having molecular weights over 23 kDa. The anti-89 kDa enamelin serum recognized the 89 kDa enamelin and lower molecular weight proteins, but neither the amelogenins nor the 13-17 kDa nonamelogenins. The antiserum against the 13-17 kDa nonamelogenins showed no cross reactivity to the 89 kDa enamelin, but recognized higher molecular weight nonamelogenins. In immunohisto-chemical preparations of the porcine tooth germs, the 25 kDa amelogenin-like immunoreactivity over immature enamel decreased in a gradient from the enamel surface to the middle layer. In the inner layer immunoreactivity was concentrated over the prism sheaths. The C-terminal specific 25 kDa amelogenin-like immunoreactivity was intense at the outer layer of immature enamel and decreased sharply toward the middle layer. Prism sheaths were intensely stained by the antiserum to the 13-17 kDa nonamelogenins. The 89 kDa enamelin-like immunoreactivity over enamel prisms was intense at the outer layer and decreased toward the middle layer. Staining by the anti-89 kDa enamelin serum of prism sheaths was faint. In immature rat incisor enamel, the C-terminal specific 25 kDa amelogenin antiserum demonstrated a staining pattern similar to that in the immature enamel of the pig. Distinct 13-17 kDa nonamelogenin-like and 89 kDa enamelin-like immunoreactivities were found especially in the layer adjacent to the Tomes' process. We conclude that some enamel proteins are degraded soon after their secretion from the secretory ameloblast in the rat and the pig. The specific enamel proteins which reacted with the antiserum to the 13-17 kDa nonamelogenins seem to be involved with the formation of prism sheaths in immature porcine enamel, but not in rat incisor enamel.