Reymond I, Almarghini K, Tappaz M
INSERM U 171/CNRS URA 1195, Centre Hospitalier Lyon-Sud, Pierre Benite, France.
Neuroscience. 1996 Nov;75(2):619-33. doi: 10.1016/0306-4522(96)00256-4.
Immunocytochemistry of cysteine sulfinate decarboxylase was performed with a new rabbit antiserum that we have recently produced and characterized using as antigen an 11,000-fold purified fraction isolated from rat liver. This antiserum precipitated cysteine sulfinate decarboxylase enzymatic activity, labeled one band (mol. wt 51,000) on immunoblots of crude tissue extracts and did not stain any cells in peripheral tissues devoid of cysteine sulfinate decarboxylase. According to these criteria, this antiserum appeared to be specific for cysteine sulfinate decarboxylase. Numerous cells were immunolabeled in the cerebellum and the hippocampus. Most notable was the labeling of the small cells surrounding the Purkinje cells and sending radial fibers up to the pial surface of the cerebellar cortex or the staining of small star-shaped cells with thin immunolabeled processes abutting on blood vessels. Identified nerve cells such as the Purkinje cells and granule cells in the cerebellum or the pyramidal and granule cells in the hippocampus were devoid of any immunoreactivity. simultaneous double immunofluorescence was carried out using anti-glial fibrillary acidic protein or anti-S-100 monoclonal antibodies. Cysteine sulfinate decarboxylase as well as glial fibrillary acidic protein- or S-100-immunopositive cells were plotted independently for the same section. Quantitative analysis of the maps indicated that the overwhelming majority of cysteine sulfinate decarboxylase-immunolabeled cells were positive for the established astrocytes markers, glial fibrillary acidic protein or S-100. Between 82 and 98% of cysteine sulfinate decarboxylase-immunolabeled cells were also glial fibrillary acidic protein-positive, depending upon the layer. Cysteine sulfinate decarboxylase immunostaining was localized within the cytoplasm, while that of glial fibrillary acidic protein was linked to the cytoskeleton. Since both labels could not be fully superposed, some double immunolabeled cells may have escaped our analysis. More than 94% up to 99% of cysteine sulfinate decarboxylase-immunolabeled cells were simultaneously S-100-immunopositive. Our quantitative data establish that cysteine sulfinate decarboxylase is strictly localized in astrocytes in the cerebellum and in the hippocampus. This finding suggests that taurine is synthesized by astrocytes in the brain and accordingly may play a role in relation to glial function, possibly within the framework of glial-neuronal interactions.
使用我们最近制备并鉴定的一种新的兔抗血清，对半胱氨酸亚磺酸脱羧酶进行了免疫细胞化学分析。该抗血清以从大鼠肝脏中分离出的纯化了11000倍的组分作为抗原。这种抗血清沉淀了半胱氨酸亚磺酸脱羧酶的酶活性，在粗组织提取物的免疫印迹上标记出一条带（分子量51000），并且在缺乏半胱氨酸亚磺酸脱羧酶的外周组织中未对任何细胞进行染色。根据这些标准，这种抗血清似乎对半胱氨酸亚磺酸脱羧酶具有特异性。在小脑和海马中有许多细胞被免疫标记。最显著的是围绕浦肯野细胞的小细胞的标记，这些小细胞发出放射状纤维直至小脑皮质的软膜表面，或者是对小星形细胞的染色，这些细胞有薄的免疫标记突起邻接血管。已鉴定的神经细胞，如小脑中的浦肯野细胞和颗粒细胞或海马中的锥体细胞和颗粒细胞，没有任何免疫反应性。使用抗胶质纤维酸性蛋白或抗S-100单克隆抗体进行了同步双重免疫荧光。在同一切片上分别绘制半胱氨酸亚磺酸脱羧酶以及胶质纤维酸性蛋白或S-100免疫阳性细胞。对图谱的定量分析表明，绝大多数半胱氨酸亚磺酸脱羧酶免疫标记的细胞对已确定的星形胶质细胞标记物胶质纤维酸性蛋白或S-100呈阳性。根据不同的层，82%至98%的半胱氨酸亚磺酸脱羧酶免疫标记细胞也是胶质纤维酸性蛋白阳性。半胱氨酸亚磺酸脱羧酶免疫染色定位于细胞质内，而胶质纤维酸性蛋白的免疫染色与细胞骨架相关。由于两种标记不能完全重叠，一些双重免疫标记的细胞可能未被我们分析到。超过94%至99%的半胱氨酸亚磺酸脱羧酶免疫标记细胞同时也是S-100免疫阳性。我们的定量数据表明，半胱氨酸亚磺酸脱羧酶严格定位于小脑和海马中的星形胶质细胞中。这一发现表明，牛磺酸是由脑中的星形胶质细胞合成的，因此可能在胶质细胞功能方面发挥作用，可能是在胶质-神经元相互作用的框架内。
