Lavi E, Wang Q, Stieber A, Gonatas N K
Department of Pathology and Laboratory Medicine, University of Pennsylvania, School of Medicine, Philadelphia 19104-6079.
Brain Res. 1994 Jun 6;647(2):273-85. doi: 10.1016/0006-8993(94)91327-7.
The Golgi apparatus-complex (GA), is a key organelle involved in several posttranslational modifications of polypeptides destined for lysosomes, plasma membranes and secretion. As reported from this laboratory, certain astrocytes in rat brain contain cisternae of the GA not only in perikarya, but also in processes. In order to further investigate which type of astrocytes contain GA in processes we conducted the present study using primary cultures of rat astrocytes and organelle specific antibodies against the GA and the rough endoplasmic reticulum (RER). While the perikarya of all cells contained elements of the GA, only a single process of a subset of type I astrocytes, negative to antibodies A2B5 and HNK-1, contained GA. In contrast, elements of the RER were found within perikarya and all processes. In order to confirm that the immunostained structures in processes indeed represent the GA, we exposed cultures to Brefeldin A (BFA), a secretion blocker which disperses the GA and redistributes it to the RER. We observed that BFA disrupted the GA of both perikarya and processes. However, astrocytes were resistant to prolonged incubations with BFA, while a similar treatment killed cultured fibroblasts and PC-12 cells. Furthermore, in astrocytes exposed to BFA for several days, the delicate network of glial fibrillary acidic protein (GFAP), was replaced by large perinuclear masses of the protein. These observations demonstrate that a subset of type I astrocytes have a single process with elements of the GA. We suggest that this specialization of the GA may be related to yet unrecognized secretory or protein processing functions of these cells. The resistance of astrocytes to BFA and the striking changes in their cytoskeleton induced by the drug, may contribute to studies on the mechanism(s) of action of BFA.
高尔基体复合体(GA)是一种关键的细胞器,参与了多种多肽的翻译后修饰,这些多肽最终会被运送至溶酶体、质膜并进行分泌。据本实验室报道,大鼠脑中的某些星形胶质细胞不仅在胞体中含有GA的扁平囊泡,在其突起中也有。为了进一步研究哪种类型的星形胶质细胞在其突起中含有GA,我们使用大鼠星形胶质细胞原代培养物以及针对GA和粗面内质网(RER)的细胞器特异性抗体进行了本研究。虽然所有细胞的胞体都含有GA成分,但只有一部分I型星形胶质细胞的单个突起含有GA,这些I型星形胶质细胞对抗体A2B5和HNK - 1呈阴性反应。相比之下,RER成分则在胞体和所有突起中均有发现。为了证实突起中免疫染色的结构确实代表GA,我们将培养物暴露于布雷菲德菌素A(BFA),一种分泌阻断剂,它会使GA分散并重新分布至RER。我们观察到BFA破坏了胞体和突起中的GA。然而,星形胶质细胞对长时间用BFA孵育具有抗性,而类似的处理会杀死培养的成纤维细胞和PC - 12细胞。此外,在暴露于BFA数天的星形胶质细胞中,胶质纤维酸性蛋白(GFAP)的精细网络被该蛋白的大核周团块所取代。这些观察结果表明,一部分I型星形胶质细胞有一个含有GA成分的单个突起。我们认为,GA的这种特化可能与这些细胞尚未被认识的分泌或蛋白质加工功能有关。星形胶质细胞对BFA的抗性以及药物诱导的其细胞骨架的显著变化,可能有助于对BFA作用机制的研究。