Kawasaki Takahiko, Takagi Yoshiko, Yamatani Hitoshi, Hirata Tatsumi
Division of Brain Function, National Institute of Genetics, Graduate University for Advanced Studies, Yata 1111, Mishima 411-8540, Japan.
J Neurobiol. 2005 Feb 15;62(3):330-40. doi: 10.1002/neu.20105.
During development, olfactory bulb axons navigate a complex microenvironment composed of myriad molecules to construct a bundle called the lateral olfactory tract. The axons themselves also express thousands of different molecules. In the present study, we produced and characterized six monoclonal antibodies that label the lateral olfactory tract and its surroundings in a unique pattern. The labeling profiles suggested that the antigen molecules recognized by each antibody are heterogeneously distributed around the developing lateral olfactory tract. We developed an efficient screening method to identify the antigen molecules by combining expression of a cDNA library in COS-7 cells and the subsequent immunohistochemical staining of the cells. The systematic screening successfully identified specific cDNA clones for all of the monoclonal antibodies, which highly probably coded for the antigen molecules, and therefore unveiled the molecular nature of local components that embrace the developing lateral olfactory tract in mice.
在发育过程中,嗅球轴突在由无数分子组成的复杂微环境中导航,以构建一个称为外侧嗅束的束状结构。轴突本身也表达数千种不同的分子。在本研究中,我们制备并鉴定了六种单克隆抗体,它们以独特的模式标记外侧嗅束及其周围环境。标记图谱表明,每种抗体识别的抗原分子在发育中的外侧嗅束周围呈异质性分布。我们开发了一种有效的筛选方法,通过将cDNA文库在COS-7细胞中的表达与随后对细胞的免疫组织化学染色相结合来鉴定抗原分子。系统筛选成功地为所有单克隆抗体鉴定出了特定的cDNA克隆,这些克隆极有可能编码抗原分子,从而揭示了包围小鼠发育中外侧嗅束的局部成分的分子性质。