LOCALIZATION OF CATHEPSINS-B, CATHEPSINS-D, AND CATHEPSINS-L IN THE RAT OSTEOCLAST BY IMMUNO-LIGHT AND IMMUNOELECTRON MICROSCOPY
LOCALIZATION OF CATHEPSINS-B, CATHEPSINS-D, AND CATHEPSINS-L IN THE RAT OSTEOCLAST BY IMMUNO-LIGHT AND IMMUNOELECTRON MICROSCOPY
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DOI：
10.1007/bf00315829
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发表时间：
1994-01-01
期刊：
影响因子：
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通讯作者：
TANAKA, T
中科院分区：
文献类型：
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作者：
GOTO, T;KIYOSHIMA, T;TANAKA, T
用免疫光镜和电镜技术，用抗生物素蛋白-生物素-过氧化物酶复合物（ABC）法研究了大鼠破骨细胞中组织蛋白酶B、D和L的定位。在冷冻切片制备的光学显微镜，组织蛋白酶D的免疫反应被发现在许多囊泡和空泡，但没有检测到沿着破骨细胞的吸收陷窝。组织蛋白酶B和L的免疫反应强烈沿着陷窝，只有弱的细胞内免疫反应观察到的小泡和周围部分的空泡附近的皱褶边缘。在未与抗体孵育的对照切片中，在破骨细胞中或沿破骨细胞的吸收陷窝沿着未发现组织蛋白酶。在电子显微镜水平上，在许多空泡和囊泡中发现了强的细胞内组织蛋白酶D反应性，而细胞外组织蛋白酶D仅在皱褶边缘的基部轻微检测到，但在侵蚀的骨基质中未发现。大多数破骨细胞显示出强烈的细胞外沉积的组织蛋白酶B和L的胶原纤维和骨基质下的皱褶边缘。组织蛋白酶L的细胞外沉积强于组织蛋白酶B。此外，组织蛋白酶B和L免疫标记了一些小凹和部分壶腹细胞外间隙，在切片中显示为空泡。相反，细胞内的组织蛋白酶B和L的反应是弱的：含组织蛋白酶的囊泡和空泡作为初级和次级溶酶体只发生稀疏。这些发现表明，与组织蛋白酶D不同，组织蛋白酶B和L迅速释放到细胞外基质中，并参与皱褶边缘尖端附近含有胶原纤维的有机骨基质的降解。组织蛋白酶L可能比组织蛋白酶B更有效地降解骨基质。
The localization of cathepsins B, D, and L was studied in rat osteoclasts by immuno-light and -electron microscopy using the avidin-biotin-peroxidase complex (ABC) method. In cryosections prepared for light microscopy, immunoreactivity for cathepsin D was found in numerous vesicles and vacuoles but was not detected along the resorption lacunae of osteoclasts. However, immunoreactivity for cathepsins B and L occurred strongly along the lacunae, and only weak intracellular immunoreactivity was observed in the vesicles and peripheral part of the vacuoles near the ruffled border. In control sections that were not incubated with the antibody, no cathepsins were found in the osteoclasts or along the resorption lacunae of osteoclasts. At the electron microscopic level, strong intracellular reactivity of cathepsin D was found in numerous vacuoles and vesicles, while extracellular cathepsin D was only slightly detected at the base of the ruffled border but was not found in the eroded bone matrix. Most osteoclasts showed strong extracellular deposition of cathepsins B and L on the collagen fibrils and bone matrix under the ruffled border. The extracellular deposition was stronger for cathepsin L than for cathepsin B. Furthermore cathepsins B and L immunolabled some pits and part of the ampullar extracellular spaces, appearing as vacuoles in the sections. Conversely, the intracellular reactivity for cathepsins B and L was weak: cathepsin-containing vesicles and vacuoles as primary and secondary lysosomes occurred only sparsely. These findings suggest that cathepsins B and L, unlike cathepsin D, are rapidly released into the extracellular matrix and participate in the degradation of organic bone matrix containing collagen fibrils near the tip of the ruffled border. Cathepsin L may be more effective in the degradation of bone matrix than cathepsin B.