Immunocytochemical localization of cyclic GMP, cGMP-dependent protein kinase, calmodulin and calcineurin in Paramecium tetraurelia.
Immunocytochemical localization of cyclic GMP, cGMP-dependent protein kinase, calmodulin and calcineurin in Paramecium tetraurelia.
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四尿草草履虫中环 GMP、cGMP 依赖性蛋白激酶、钙调蛋白和钙调神经磷酸酶的免疫细胞化学定位。
DOI：
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发表时间：
1983
影响因子：
6.6
通讯作者：
J. Schultz
中科院分区：
文献类型：
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作者：
S. Klumpp;A. Steiner;J. Schultz
已经用免疫细胞化学方法检查了草履虫细胞中 cGMP、cGMP 依赖性蛋白激酶、钙调蛋白和钙调蛋白结合蛋白钙调神经磷酸酶的定位。这些分子似乎在很大程度上位于这种原生动物的纤毛中。为了确定抗体能够进入所有细胞区室，我们使用了三种不同的免疫细胞化学制剂：（i）使用“全细胞”制剂，四种分子的免疫荧光染色主要在纤毛中可见； (ii) 在“去纤毛”草履虫中，在细胞表面以规则图案发现 cGMP 和钙调蛋白染色，最有可能代表运动体； (iii)使用“切片细胞”，额外的细胞质钙调蛋白似乎与糖原颗粒相关，如与α-淀粉酶预孵育后颗粒染色图案消失所证明的那样。相比之下，cGMP、cGMP依赖性蛋白激酶和钙调神经磷酸酶的荧光非常弱并且在细胞体内弥散。用任何抗体染色后均未检测到核荧光。由于草履虫纤毛中的 cGMP、cGMP 依赖性蛋白激酶、鸟苷酸环化酶-钙调蛋白复合物和钙调磷酸酶共定位，因此讨论了这些成分参与纤毛活动调节的情况。
The localization of cGMP, cGMP-dependent protein kinase, calmodulin and the calmodulin-binding protein calcineurin in Paramecium tetrauelia cells has been examined with immunocytochemical methods. These molecules appeared to be localized to a large extent in the cilia of this protozoan. To ascertain that antibodies had access to all cellular compartments we have used three different preparations for immunocytochemistry: (i) with 'whole cell' preparations immunofluorescent staining for the four molecules was mainly visible in the cilia; (ii) in 'deciliated' Paramecium, staining for cGMP and calmodulin was found in regular patterns on the cell surface most likely representing kinetosomes; (iii) using 'sectioned cells', additional cytoplasmic calmodulin appeared to be associated with glycogen particles as evidenced by the disappearance of the granular staining pattern after preincubation with alpha-amylase. In contrast, cGMP, cGMP-dependent protein kinase and calcineurin fluorescence was only very weak and diffuse in cell bodies. No nuclear fluorescence was detectable after staining with any of the antibodies. Because of the colocalization of cGMP, cGMP-dependent protein kinase, a guanylate cyclase-calmodulin-complex, and calcineurin in cilia from Paramecium, an involvement of these components in the regulation of ciliary activity is discussed.