Tazaki A, Gaudieri S, Ikeo K, Gojobori T, Watanabe K, Agata K
Department of Life Science, Himeji Institute of Technology, Hyogo, 678-1297, Japan.
Biochem Biophys Res Commun. 1999 Jul 5;260(2):426-32. doi: 10.1006/bbrc.1999.0933.
In order to investigate the neural connection of planarian, it is imperative to produce an antibody that specifically stains axons. To identify axon-specific genes, we constructed a cDNA library from a single eye by using a single cell PCR method, in which visual neurons are major components, and sequenced one thousand independent clones. We succeeded in the identification of a planarian homologue of synaptotagmin, Djsyt, whose specific expression in neurons was confirmed by in situ hybridization. The antibody against DjSYT specifically stained axons although its mRNA is distributed in the cell bodies. By using anti-DjSYT, we succeeded in the visualization of neural connections in planarians by whole mount staining. The anti-DjSYT antibody will become a powerful tool to analyze the molecular mechanisms underlying neural network formation in planarian.
为了研究涡虫的神经连接，制备一种能特异性标记轴突的抗体至关重要。为了鉴定轴突特异性基因，我们采用单细胞PCR方法从单眼中构建了一个cDNA文库（其中视觉神经元是主要成分），并对一千个独立克隆进行了测序。我们成功鉴定出了突触结合蛋白的涡虫同源物DjSYT，其在神经元中的特异性表达通过原位杂交得以证实。尽管DjSYT的mRNA分布在细胞体中，但针对DjSYT的抗体却能特异性地标记轴突。通过使用抗DjSYT抗体，我们成功地通过整体染色观察到了涡虫的神经连接。抗DjSYT抗体将成为分析涡虫神经网络形成潜在分子机制的有力工具。
