[Simultaneous demonstration of glutamate decarboxylase and synaptophysin in paraffin sections of rat cerebellum].

D E Korzhevskiy, Ye G Gilerovich, O V Kirik, O S Alekseyeva, I P Grigoriyev
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Abstract

The article presents highly reproducible and inexpensive protocol for simultaneous demonstration of glutamate decarboxylase (GAD67), the key enzyme of gamma-aminobutyric acid (GABA) synthesis and synaptophysin (SYP), a marker protein of synaptic vesicles using confocal laser microscopy. In the cerebellar cortex, GAD labels Purkinje cells and pinceaux in their basal parts and is unevenly distributed in the neuropil of molecular and granular layers. SYP clearly marks the contours of large dendrites of Purkinje cells in molecular layer, while in the granular layers it labels parts of cerebellar glomeruli--the terminals of the mossy fibers. GAD-immunopositive structures (GABA-ergic axons of stellate cells--Golgi cells) are often located at periphery of the glomeruli. In the peripheral zone of the glomeruli, colocalization of GAD- and SYP-immunopositive structures was observed, suggesting the presence of GABA-ergic synapses in this zone.

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[在大鼠小脑石蜡切片上同时展示谷氨酸脱羧酶和突触素]。
本文提出了一种高重复性和低成本的方案,用于同时展示γ -氨基丁酸(GABA)合成的关键酶谷氨酸脱羧酶(GAD67)和突触囊泡标记蛋白突触素(SYP)。在小脑皮层,GAD标记浦肯野细胞和针状细胞在其基部,在分子层和颗粒层的神经层中分布不均匀。SYP在分子层清晰地标记浦肯野细胞大树突的轮廓,而在颗粒层则标记部分小脑肾小球——苔藓纤维的末端。gad免疫阳性结构(星状细胞-高尔基细胞的gaba能轴突)通常位于肾小球的周围。在肾小球外周区,观察到GAD和syp免疫阳性结构的共定位,表明该区域存在gaba能突触。
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