Phylogeny and comparative anatomy of the cholinergic neuron in invertebrate and vertebrates

胆碱乙酰转移酶（Choline acetyltransferase, ChAT）是胆碱能神经元最特异的标志物。利用分子生物学技术制备了大鼠ChAT抗血清，并建立了免疫组化反应。最近，我们发现抗血清在石蜡包埋切片中染色胆碱能神经元，这使我们能够研究胆碱能神经元的系统发育和比较解剖学。然而，我们发现，抗血清的跨物种反应性不够广泛，不足以完成本项目。因此，我们生产了一种抗水泡乙酰胆碱转运蛋白（VAChT）的血清，这是胆碱能神经元的另一种特异性蛋白质，并检查它是否可以用于本项目。为此，利用抗sear - to和地高辛水平RNA对ChAT和VAChT的证明，我们通过免疫组织化学和原位杂交组织化学广泛比较了ChAT和VAChT mrna和相应蛋白的分布，无论是单独的还是与它们的各种组合，证明抗VAChT血清可以用于本项目。

Choline acetyltransferase (ChAT), the biosynthetic enzyme for ACh, is the most specific marker for the cholinergic neuron. Using the molecular biological techniques, we previously produced an antiserum to rat ChAT and established the immunohistochemistry. Recently we found that the antiserum stains cholinergic neurons in paraffin-embedded sections, and this enables us to study the phylogeny and comparative anatomy of the cholinergic neuron. We found, hoever, that cross-species reactivity of the antiserum is not wide enough to complete the present project. We, therefore, produced an antiserum to vesicular ACh transporter (VAChT), another specific protein for the cholinergic neuron, and examined whether or not it can be used for the present project. For this, using antisear to and digoxigenin-laveled RNA proves for ChAT and VAChT,we compared extensively the distribution of mRNAs and the corresponding proteins for ChAT and VAChT by immunohistochemistry and in situ hybridization histochemistry, both alone and with various combinations of them, and demonstrated that the antiserum to VAChT can be used for the present project.