Oofusa Ken, Yoshizato Katsutoshi
Developmental Biology Laboratory, Department of Biology, Faculty of Science, Tokyo Metropolitan University, 2-1-1 Fukazawa, Setagaya-ku, Tokyo 158, Japan.
Molecular Cell Science Laboratory, Zoological Institute, Faculty of Science, Hiroshima University, 1-1-89 Higashisenda-machi, Naka-ku, Hiroshima 730, Japan.
Dev Growth Differ. 1991 Aug;33(4):329-339. doi: 10.1111/j.1440-169X.1991.00329.x.
Tissue inhibitor of metalloproteinases (TIMP, a specific inhibitor of collagenase) was found to inhibit thyroid hormone-induced tail regression, suggesting the important role of collagenase in this process. Collagenase was purified from culture media of back skin of tadpole of bullfrog, Rana catesbeiana. Anti-tadpole collagenase polyclonal antisera were obtained against the purified enzyme. The antibody inhibited the activity of tadpole collagenase. The antisera reacted to tissues of adult bullfrogs, tadpoles of african clawed frog, Xenopus laevis, and adult newts, Cynopus pyrrhogaster, and also reacted to human fibroblast collagenase. Immunoblot analyses suggested that tadpole collagenase lacks the procollagenase which is generally found in mammalian collagenases. Intense immunological stains were observed for the tissues of thyroid hormone-treated tadpoles as compared to those of untreated animals. Thyroid hormone increased amounts of collagenase not only in epidermal layer but also in mesenchymal tissues including fibroblastic cells.
金属蛋白酶组织抑制剂（TIMP，一种胶原酶的特异性抑制剂）被发现可抑制甲状腺激素诱导的尾部退化，这表明胶原酶在该过程中起重要作用。胶原酶是从牛蛙（Rana catesbeiana）蝌蚪背部皮肤的培养基中纯化得到的。针对纯化后的酶制备了抗蝌蚪胶原酶多克隆抗血清。该抗体抑制了蝌蚪胶原酶的活性。该抗血清可与成年牛蛙、非洲爪蟾（Xenopus laevis）蝌蚪以及成年蝾螈（Cynopus pyrrhogaster）的组织发生反应，也可与人成纤维细胞胶原酶发生反应。免疫印迹分析表明，蝌蚪胶原酶缺乏通常在哺乳动物胶原酶中发现的前胶原酶。与未处理动物的组织相比，在甲状腺激素处理的蝌蚪组织中观察到强烈的免疫染色。甲状腺激素不仅增加了表皮层中胶原酶的含量，还增加了包括成纤维细胞在内的间充质组织中胶原酶的含量。
