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新型抗环磷酸鸟苷单克隆抗体及其在大鼠脑免疫细胞化学中的应用。

Novel type of monoclonal antibodies against cyclic GMP and application to immunocytochemistry of the rat brain.

作者信息

Sakaue M, Kuno T, Tanaka C

机构信息

Department of Pharmacology, Kobe University School of Medicine, Japan.

出版信息

Jpn J Pharmacol. 1988 Sep;48(1):47-56. doi: 10.1254/jjp.48.47.

Abstract

A novel type of monoclonal antibodies against cyclic GMP were produced to study the immunocytochemical distribution of cyclic GMP in the rat brain. Cyclic GMP conjugated to bovine serum albumin with glutaraldehyde was used as an immunogen, and monoclonal antibodies were produced. The one monoclonal antibody which did not crossreact against other nucleotides was applied to the immunocytochemistry of the rat brain. Cyclic GMP immunoreactivities were distributed unevenly in the rat brain. The cerebellar cortex, hippocampus and cerebral cortex contained a high degree of cyclic GMP immunoreactivity, while most of the white matter was not stained. In the cerebellar cortex, stellate cells and Golgi cells showed intense immunoreactivities, but granule cells showed weak immunoreactivities. Approximately 60-80% of the Purkinje cells showed intense immunoreactivities, while the remaining ones showed only weak staining. The pyramidal cells in the cerebral cortex and hippocampus also showed intense immunostaining. Some glial cells adjacent to the Purkinje cells also stained. The nuclei of cyclic GMP-immunoreactive cells were not stained. These immunocytochemical distributions of cyclic GMP are in fairly good agreement with reported the biochemical data and the immunocytochemical distribution of guanylate cyclase. These monoclonal antibodies should be helpful for elucidating the physiological role of cyclic GMP in the brain.

摘要

为研究环磷酸鸟苷(cGMP)在大鼠脑中的免疫细胞化学分布,制备了一种新型的抗cGMP单克隆抗体。用戊二醛将cGMP与牛血清白蛋白偶联作为免疫原,制备单克隆抗体。将一种不与其他核苷酸发生交叉反应的单克隆抗体应用于大鼠脑的免疫细胞化学研究。cGMP免疫反应性在大鼠脑中分布不均。小脑皮质、海马和大脑皮质含有高度的cGMP免疫反应性,而大部分白质未被染色。在小脑皮质中,星状细胞和高尔基细胞显示出强烈的免疫反应性,而颗粒细胞显示出较弱的免疫反应性。约60%-80%的浦肯野细胞显示出强烈的免疫反应性,其余的仅显示弱阳性染色。大脑皮质和海马中的锥体细胞也显示出强烈的免疫染色。一些与浦肯野细胞相邻的神经胶质细胞也被染色。cGMP免疫反应性细胞的细胞核未被染色。cGMP的这些免疫细胞化学分布与已报道的生化数据和鸟苷酸环化酶的免疫细胞化学分布相当一致。这些单克隆抗体有助于阐明cGMP在脑中的生理作用。

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