Localization of the O-GlcNAc transferase and O-GlcNAc-modified proteins in rat cerebellar cortex
Localization of the O-GlcNAc transferase and O-GlcNAc-modified proteins in rat cerebellar cortex
复制标题
DOI：
10.1016/s0006-8993(02)04158-6
复制
发表时间：
2003-03-21
期刊：
影响因子：
2.9
通讯作者：
Hart, GW
中科院分区：
文献类型：
--
作者：
Akimoto, Y;Comer, FI;Hart, GW
O-连接的N-乙酰葡糖胺（O-GlcNAc）是一种普遍存在的核质蛋白修饰，其与细胞骨架蛋白、信号蛋白和转录因子上的磷酸化具有复杂的相互作用。O-GlcNAc在单细胞水平上对生命至关重要，许多间接证据表明它在神经细胞生物学和神经退行性疾病中起重要作用。在这里，我们显示了O-GlcNAc转移酶（OGTase）mRNA，OGTase蛋白，和O-GlcNAc修饰的蛋白质在大鼠小脑皮质的本地化。用原位杂交组织化学方法检测OGTase mRNA的表达部位。在神经元，尤其是浦肯野细胞中存在强烈的杂交信号。抗OGTase的荧光标记抗体染色几乎所有的神经元，特别是强烈的反应，在浦肯野细胞，其中细胞核，核周体，和树突染色最强烈。使用免疫电镜标记，OGTase被认为是丰富的常染色质，在细胞质基质，在神经末梢，和周围的微管树突。在神经末梢，免疫金标记观察到周围的突触囊泡，与酶更密集地定位在突触前比突触后的终端。使用O-GlcNAc抗体，我们发现糖定位反映了OGTase的结果。总的来说，这些数据支持O-GlcNAc在脑细胞关键过程中的假设作用，包括转录调节、突触囊泡分泌、转运和信号转导。因此，通过调节磷酸化或蛋白质协会的关键监管和细胞骨架蛋白，O-GlcNAc可能是重要的小脑的许多功能。(C)2002 Elsevier Science B. V.保留所有权利。
O-linked N-acetylglucosamine (O-GlcNAc) is a ubiquitous nucleocytoplasmic protein modification that has a complex interplay with phosphorylation on cytoskeletal proteins, signaling proteins and transcription factors. O-GlcNAc is essential for life at the single cell level, and much indirect evidence suggests it plays an important role in nerve cell biology and neurodegenerative disease. Here we show the localization of O-GlcNAc Transferase (OGTase) mRNA, OGTase protein, and O-GlcNAc-modified proteins in the rat cerebellar cortex. The sites of OGTase mRNA expression were determined by in situ hybridization histochemistry. Intense hybridization signals were present in neurons, especially in the Purkinje cells. Fluorescent-tagged antibody against OGTase stained almost all of the neurons with especially intense reactivity in Purkinje cells, within which the nucleus, perikaryon, and dendrites were most intensely stained. Using immuno-electron microscopic labeling, OGTase was seen to be enriched in euchromatin, in the cytoplasmic matrix, at the nerve terminal, and around microtubules in dendrites. In nerve terminals, immuno-gold labeling was observed around synaptic vesicles, with the enzyme more densely localized in the presynaptic terminals than in the postsynaptic ones. Using an antibody to O-GlcNAc, we found the sugar localizations reflected results seen for OGTase. Collectively, these data support hypothesized roles for O-GlcNAc in key processes of brain cells, including the regulation of transcription, synaptic vesicle secretion, transport, and signal transduction. Thus, by modulating the phosphorylation or protein associations of key regulatory and cytoskeletal proteins, O-GlcNAc is likely important to many functions of the cerebellum. (C) 2002 Elsevier Science B.V. All rights reserved.