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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >Characterization of tyramine beta-hydroxylase in planarian Dugesia japonica: cloning and expression.
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Characterization of tyramine beta-hydroxylase in planarian Dugesia japonica: cloning and expression.

机译：涡虫日本uge中酪胺β-羟化酶的表征：克隆和表达。

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The planarian Dugesia japonica has a relatively well-organized central nervous system (CNS) consisting of a brain and ventral nerve cords (VNCs), and can completely regenerate it CNS utilizing pluripotent stem cells present in the mesenchymal space. This remarkable capacity has begun to be exploited for research on neural regeneration. Recently, several kinds of molecular markers for labeling of neural subtypes have been reported in planarians. These molecular markers are useful for visualizing the distinct neural populations in planarians. In this study, we isolated a cDNA encoding tyramine beta-hydroxylase (TBH), an octopamine (OA) biosynthetic enzyme, by degenerate PCR in the planarian D. japonica, and named it DjTBH (D. japonica tyramine beta-hydroxylase). In order to examine whether DjTBH contributes to OA biosynthesis, we measured the OA content in DjTBH-knockdown planarians created by RNA interference. In addition, to examine the specificity of DjTBH for OA biosynthesis, we measured not only OA content but also noradrenaline (NA) content, because NA is synthesized by a pathway similar to that for OA. According to high-performance liquid chromatography analysis, the amount of OA, but not NA, was significantly decreased in DjTBH-knockdown planarians. In addition, we produced anti-DjTBH antibody to visualize the octopaminergic neural network. As shown by immunofluorescence analysis using anti-DjTBH antibody, DjTBH-immunopositive neurons were mainly distributed in the head region, and elongated their dendrites and/or axons along the VNCs. In order to visualize octopaminergic and dopaminergic nervous systems (phenolamine/catecholamine nervous system) in the planarian CNS, double-immunofluorescence analysis was carried out using both anti-DjTBH antibody and anti-DjTH (a planarian tyrosine hydroxylase) antibody. DjTBH-immunopositive neurons and DjTH-immunopositive neurons mainly formed distinct neural networks in the head region. Here, we demonstrated that DjTBH clearly contributes to OA biosynthesis, and DjTBH antibody is a useful tool for detecting octopaminergic neurons in planarians.
机译：涡虫日本杜鹃具有相对组织良好的中枢神经系统（CNS），由脑和腹神经索（VNC）组成，并且可以利用间充质空间中存在的多能干细胞完全再生CNS。这种非凡的能力已开始用于神经再生研究。最近，在平面学家中已经报道了几种标记神经亚型的分子标记。这些分子标记可用于可视化涡虫中不同的神经种群。在这项研究中，我们通过在涡虫D. japonica中进行简并PCR分离了编码酪胺β-羟化酶（TBH），一种章鱼胺（OA）生物合成酶的cDNA，并将其命名为DjTBH（D. japonica酪胺β-羟化酶）。为了检查DjTBH是否有助于OA生物合成，我们测量了由RNA干扰产生的DjTBH-nockdown平面虫中的OA含量。此外，为了检查DjTBH对OA生物合成的特异性，我们不仅测量了OA含量，还测量了去甲肾上腺素（NA）含量，因为NA是通过类似于OA的途径合成的。根据高效液相色谱分析，在DjTBH击倒型涡旋仪中OA的量显着减少，而NA则没有。此外，我们生产了抗DjTBH抗体以可视化章鱼碱能神经网络。如使用抗DjTBH抗体的免疫荧光分析所示，DjTBH免疫阳性神经元主要分布在头部区域，并沿VNC拉长其树突和/或轴突。为了可视化在涡虫中枢神经系统中的八胺能和多巴胺能神经系统（酚胺/儿茶酚胺神经系统），使用抗DjTBH抗体和抗DjTH（涡虫酪氨酸羟化酶）抗体进行了双重免疫荧光分析。 DjTBH免疫阳性神经元和DjTH免疫阳性神经元主要在头部区域形成独特的神经网络。在这里，我们证明了DjTBH显然有助于OA生物合成，并且DjTBH抗体是检测涡虫中章鱼胺能神经元的有用工具。

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