Cajaraville M P, Völkl A, Fahimi H D
Department of Cell Biology and Morphological Sciences, University of the Basque Country, Bilbo/Spain.
Eur J Cell Biol. 1992 Dec;59(2):255-64.
The present study was undertaken because of the paucity of information on peroxisomes in molluscs and the increasing importance of these organisms as sensitive indicators of environmental pollution. Peroxisomes were identified by electron microscopy in all three main cell types of the digestive gland of the bivalve mollusc Mytilus galloprovincialis Lmk. They stained weakly with the alkaline diaminobenzidine reaction but showed distinct immunolabeling with an antibody against mammalian catalase by the postembedding protein A-gold procedure. In addition, mussel digestive gland peroxisomes were isolated by differential and metrizamide-density gradient centrifugation, and a 30-fold enrichment of catalase and a 20-fold enrichment of palmitoyl-CoA oxidase was obtained over the initial homogenate. By Western blotting, isolated peroxisomes crossreacted with antibodies to catalase and, furthermore, specific and prominent labeling of isolated peroxisomes was also demonstrated in thin sections incubated with anti-catalase antibodies. These observations establish that peroxisomes in molluscan digestive gland contain the peroxisomal marker enzymes catalase and acyl-CoA oxidase and that they can be labeled by cytochemical and immunocytochemical techniques. Further studies of alterations of molluscan peroxisomes by environmentally relevant xenobiotics are warranted.
由于关于软体动物过氧化物酶体的信息匮乏,且这些生物作为环境污染敏感指标的重要性日益增加,因此开展了本研究。通过电子显微镜在双壳贝类贻贝(Mytilus galloprovincialis Lmk)消化腺的所有三种主要细胞类型中鉴定出了过氧化物酶体。它们在用碱性二氨基联苯胺反应染色时颜色较浅,但通过包埋后蛋白A-金法用抗哺乳动物过氧化氢酶抗体显示出明显的免疫标记。此外,通过差速离心和甲泛影酰胺密度梯度离心分离出贻贝消化腺过氧化物酶体,与初始匀浆相比,过氧化氢酶富集了30倍,棕榈酰辅酶A氧化酶富集了20倍。通过蛋白质免疫印迹法,分离出的过氧化物酶体与过氧化氢酶抗体发生交叉反应,此外,在用抗过氧化氢酶抗体孵育的薄片中也显示出分离出的过氧化物酶体有特异性且明显的标记。这些观察结果表明,软体动物消化腺中的过氧化物酶体含有过氧化物酶体标记酶过氧化氢酶和酰基辅酶A氧化酶,并且它们可以通过细胞化学和免疫细胞化学技术进行标记。有必要进一步研究环境相关的外源化合物对软体动物过氧化物酶体的影响。