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成年小鼠中枢神经系统中泛连接蛋白1的表达:细胞定位及4-氨基吡啶诱导癫痫发作的影响

Expression of pannexin1 in the CNS of adult mouse: cellular localization and effect of 4-aminopyridine-induced seizures.

作者信息

Zappalà A, Cicero D, Serapide M F, Paz C, Catania M V, Falchi M, Parenti R, Pantò M R, La Delia F, Cicirata F

机构信息

Department of Physiological Science, University of Catania, Viale A. Doria 6, 95125 Catania, Italy.

出版信息

Neuroscience. 2006 Aug 11;141(1):167-78. doi: 10.1016/j.neuroscience.2006.03.053. Epub 2006 May 11.

Abstract

The expression pattern of pannexin1, a gene coding for a protein that forms gap junction channels, was studied as both mRNA and protein in the CNS of adult mouse. Pannexin1 was widely expressed in the CNS by neuronal cell types but not glial cells, except for Bergmann glial cells of the cerebellar cortex. Cells positive to Ca-binding proteins, principally parvalbumin, but also calbindin and calretinin, as well as glutamate decarboxylase 67 kDa isoform, were pannexin1-positive. Pannexin1 labeling was found in cells which are known to exhibit spontaneous and synchronous discharge, such as neurons of the inferior olivary complex and the reticular thalamic nucleus, and also in neurons whose electrical activity is not coupled with neighboring cells, such as motoneurons of the spinal cord. The analysis of cellular localization showed puncta that surrounded cell bodies (e.g. the pyramidal cells of hippocampus) or restricted areas inside the cell bodies (e.g. the spinal motoneurons). In Bergmann glial cells the staining was present as fine grains that covered a large part of the cellular surface. Pannexin1 stained cells that previous studies have reported as expressing connexin36, another protein forming gap junction channels. Thus, it was possible that these two proteins could be integrated in the same functions. Since connexin36 expression levels change after seizures, we examined the expression of both pannexin1 and connexin36 in cerebral cortex, hippocampus, cerebellum and brain stem at different time intervals (2, 4 and 8 h) after i.p. injection of 4-aminopyridine, which resulted in systemic seizures. The only modification of the expression levels observed in this study concerned the progressive decrement of the connexin36 in the hippocampus, while pannexin1 expression was unchanged. This finding suggested that pannexin1 and connexin36 are involved in different functional roles or that they are expressed in different cell types and that only those expressing the Cx36 are induced to apoptosis by epileptic seizures.

摘要

泛素连接蛋白1是一种编码形成间隙连接通道蛋白的基因,研究了其在成年小鼠中枢神经系统中的mRNA和蛋白质表达模式。泛素连接蛋白1在中枢神经系统中广泛表达于神经元细胞类型,但不表达于神经胶质细胞,除了小脑皮质的伯格曼神经胶质细胞。对钙结合蛋白呈阳性的细胞,主要是小白蛋白,还有钙结合蛋白和钙视网膜蛋白,以及67 kDa亚型的谷氨酸脱羧酶,均为泛素连接蛋白1阳性。在已知表现出自发和同步放电的细胞中发现了泛素连接蛋白1标记,如下橄榄复合体和丘脑网状核的神经元,以及电活动不与相邻细胞耦合的神经元,如脊髓运动神经元。细胞定位分析显示,点状结构围绕细胞体(如海马体的锥体细胞)或细胞体内的受限区域(如脊髓运动神经元)。在伯格曼神经胶质细胞中,染色呈现为覆盖细胞表面大部分区域的细颗粒。泛素连接蛋白1染色的细胞,先前的研究报道其表达连接蛋白36,另一种形成间隙连接通道的蛋白。因此,这两种蛋白有可能整合到相同的功能中。由于癫痫发作后连接蛋白36的表达水平会发生变化,我们在腹腔注射4-氨基吡啶导致全身性癫痫发作后的不同时间间隔(2、4和8小时),检测了大脑皮质、海马体、小脑和脑干中泛素连接蛋白1和连接蛋白36的表达。本研究中观察到的表达水平的唯一变化是海马体中连接蛋白36的逐渐减少,而泛素连接蛋白1的表达未改变。这一发现表明,泛素连接蛋白1和连接蛋白36参与不同的功能作用,或者它们在不同的细胞类型中表达,并且只有那些表达Cx36的细胞会因癫痫发作而被诱导凋亡。

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