Developmentally regulated O-acetylated sialoglycans in the central nervous system revealed by a new monoclonal antibody 493D4 recognizing a wide range of O-acetylated glycoconjugates
Developmentally regulated O-acetylated sialoglycans in the central nervous system revealed by a new monoclonal antibody 493D4 recognizing a wide range of O-acetylated glycoconjugates
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DOI：
10.1023/a:1018542105832
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发表时间：
1997-11
影响因子：
3
通讯作者：
Gu Zhang;L. Ji;S. Kurono;S. Fujita;S. Furuya;Y. Hirabayashi
中科院分区：
文献类型：
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作者：
Gu Zhang;L. Ji;S. Kurono;S. Fujita;S. Furuya;Y. Hirabayashi
我们以前在大鼠胚胎大脑皮层中检测到一种碱不稳定和发育调节的抗原，该抗原可能是9- o乙酰唾液化GT3神经节苷脂（Hirabayashi Y, Hirota M, Suzuki Y, Matsumoto M, Obata K, Ando S (1989)Neurosci Lett106:193-98）。在本研究中，我们建立了一种小鼠单克隆抗体493D4，它可以识别9- o -乙酰基GT3神经节苷脂，而不能识别非o -乙酰基神经节苷脂。该抗体与9- o -乙酰基GD3的反应程度要小得多。利用该抗体，我们发现o -乙酰基GT3和o -乙酰基GD3在胎儿小鼠大脑皮层中表达强烈，出生后下降到检测不到的水平。采用493D4 tlc免疫染色结合Q-Sepharose柱层析纯化牛脑o -乙酰基神经节苷脂，结构分析表明，成人脑中o -乙酰基GD3、o -乙酰基LD1、o -乙酰基GD2和o -乙酰基GD1b的含量极低。有趣的是，抗体493D4可以检测大鼠脑组织中的to -乙酰基唾液糖蛋白。其中一种主要的免疫反应蛋白是突触素，一种特异性存在于突触囊泡中的完整膜蛋白。因此，该单克隆抗体可用于敏感检测乙酰化的神经节苷和乙酰化的唾液酸糖蛋白。
We have previously detected an alkali-labile and developmentally regulated antigen in rat embryonic cerebral cortex, which may be 9-O-acetylsialylated GT3 ganglioside (Hirabayashi Y, Hirota M, Suzuki Y, Matsumoto M, Obata K, Ando S (1989)Neurosci Lett106:193-98). In this study we established a mouse monoclonal antibody, 493D4, that recognizes 9-O-acetyl GT3 ganglioside, but not non-O-acetyl gangliosides. This antibody also reacted with 9-O-acetyl GD3 to a much lesser extent. By using this antibody, we found thatO-acetyl GT3 as well asO-acetyl GD3 were expressed strongly in fetal murine cerebral cortex and decreased to an undetectable level after birth. With the assistance of TLC-immunostaining using 493D4 together with Q-Sepharose column chromatography,O-acetyl gangliosides of bovine brain were purified and the structural analysis showed the presence ofO-acetyl GD3,O-acetyl LD1,O-acetyl GD2 andO-acetyl GD1b in the adult brain as extremely minor components. Interestingly, the antibody 493D4 could detectO-acetyl sialoglycoproteins in rat brain tissues. One of the major immunoreactive proteins was shown to be synaptophysin, an integral membrane protein specifically present in synaptic vesicles. This monoclonal antibody was therefore useful for sensitive detection of bothO-acetylated gangliosides and glycoproteins withO-acetylated sialic acids.