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首页> 外文期刊>The European Journal of Neuroscience >Localization of endoplasmic reticulum and plasma membrane Ca2+-ATPases in subcellular fractions and sections of pig cerebellum.
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Localization of endoplasmic reticulum and plasma membrane Ca2+-ATPases in subcellular fractions and sections of pig cerebellum.

机译：猪小脑亚细胞部分和切片中内质网和质膜Ca2 + -ATPase的定位

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摘要

Subcellular fractions and sections of the cerebellum were analysed to evaluate the relative activity and distribution of organellar and plasma membrane Ca2+-ATPases (SERCA and PMCA). Western blot analysis of the fractions with IID8 or Y/1F4 SERCA-specific antibodies or else with 5F10 or pbPMCA antibodies, specific to PMCA pump, revealed a major content of SERCA protein in microsomes and of PMCA protein in plasma membrane vesicles. The Ca2+-ATPase activity of microsomes was more sensitive to thapsigargin, a SERCA-specific inhibitor, whereas the activity of the plasma membrane vesicle fraction was inhibited more by vanadate, a blocker of PMCA activity. The SERCA and PMCA distribution analysed in cerebellar sections revealed IID8 antibody reactions in Purkinje cell cytoplasm, granule cells and cerebellar glomeruli. Y/1F4 gave immunostaining in Purkinje cells, molecular layer interneurons (basket and stellate cells) and glomeruli, but granule cells were not labelled. The 5F10 antibody reacted with Purkinjecells, including their dendritic spines, as well as cerebellar glomeruli, whereas the pbPMCA antibody labelled several processes in all three layers and some synaptic interaction sites. The differential content and localization of the two types of Ca2+ pumps in specific neuronal areas of pig cerebellum indicate precise Ca2+ requirements of specific cellular regions.
机译：分析小脑的亚细胞部分和切片，以评估细胞器和质膜Ca2 + -ATPases（SERCA和PMCA）的相对活性和分布。用IID8或Y / 1F4 SERCA特异性抗体，或者用5F10或pbPMCA抗体（对PMCA泵具有特异性）对级分进行蛋白质印迹分析，发现微粒体中有SERCA蛋白的主要成分，质膜囊泡中有PMCA蛋白的成分。微粒体的Ca2 + -ATPase活性对thapsigargin（一种SERCA特异性抑制剂）更为敏感，而质膜囊泡部分的活性受到钒酸盐（PMCA活性的阻断剂）的抑制作用更大。在小脑切片中分析的SERCA和PMCA分布揭示了Purkinje细胞质，颗粒细胞和小脑肾小球中的IID8抗体反应。 Y / 1F4在浦肯野细胞，分子层间神经元（篮状和星状细胞）和肾小球中进行了免疫染色，但未标记颗粒细胞。 5F10抗体与Purkinjecells包括其树突棘以及小脑肾小球发生了反应，而pbPMCA抗体则在所有三层中都标记了多个过程，并标记了一些突触相互作用位点。猪小脑特定神经元区域中两种类型的Ca2 +泵的含量差异和定位表明特定细胞区域对Ca2 +的精确需求。

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