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

无脊椎动物和脊椎动物胆碱能神经元的系统发育和比较解剖学

• 批准号: 08640854
• 负责人: ICHIKAWA Tomoyuki
• 金额: $ 1.41万
• 依托单位: Tokyo Metropolitan Institute for Neuroscience
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08640854/
• 关键词: Cholinergic neuron; Immunohistochemistry; Choline acetyltranceferase; Vesiculae acetylcholine transporter; Comparative anatomy; Phylogeny; 脊椎・無脊椎動物

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.

胆碱乙酰转移酶 (ChAT) 是乙酰胆碱的生物合成酶，是胆碱能神经元最特异的标记物。我们之前利用分子生物学技术制备了大鼠ChAT的抗血清并建立了免疫组化。最近我们发现抗血清对石蜡包埋切片中的胆碱能神经元进行染色，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，我们生产了一种针对囊泡乙酰胆碱转运蛋白（VAChT）（胆碱能神经元的另一种特异性蛋白质）的抗血清，并检查了它是否可以用于本项目。为此，我们使用抗ChAT和VAChT的RNA和地高辛标记的RNA证明，通过免疫组织化学和原位杂交组织化学，单独和它们的各种组合，广泛比较了ChAT和VAChT的mRNA和相应蛋白的分布，并证明了VAChT的抗血清可用于本项目。

项目成果

T.Ichikawa et al.: "Localization of two cholinergic markers,choline acetyltransf erase and vesicular acetylcholine transporter in the central nervous system of the rat" Journal of Chemical Neuroanatomy. 13. 23-39 (1997)

T.Ichikawa 等人：“大鼠中枢神经系统中两种胆碱能标记物、胆碱乙酰转移酶和囊泡乙酰胆碱转运蛋白的定位”《化学神经解剖学杂志》。

T.Ichikawa and T.Shimizu: "Organization of choline acetyltransferase-containing structures in the cranial nerve motor nuclei and spinal cord of the monkey" Brain Res.779. 96-103 (1998)

T.Ichikawa 和 T.Shimizu：“猴子的脑神经运动核和脊髓中含有胆碱乙酰转移酶的结构的组织”Brain Res.779。

T.Ichikawa and T.Shimizu: "Organization of choline acetyltransferase-containing structures in the cranial nerve motor nuclei and spinal cord of the monkey" Brain Research. 779. 96-103 (1998)

T.Ichikawa 和 T.Shimizu：“猴子的脑神经运动核和脊髓中含有胆碱乙酰转移酶的结构的组织”大脑研究。

T.Ichikawa et al.: "Localization of two cholinergic markers,choline acetyltransferase and vesicular acetylholine transporter in the central in the central nervous system of the rat" Journal of Chemical Neuroanatomy. (in press). (1997)

T.Ichikawa 等人：“两种胆碱能标记物、胆碱乙酰转移酶和囊泡乙酰胆碱转运蛋白在大鼠中枢神经系统中枢的定位”《化学神经解剖学杂志》。

T.Ichikawa et al.: "Localization of two cholinergic markers,choline acetyltrasferase and vesicular acetylcholine transporter in the central nervous system of the rat" Journal of Chemical Neuroanatomy. 13. 23-39 (1997)

T.Ichikawa 等人：“大鼠中枢神经系统中两种胆碱能标记物、胆碱乙酰转移酶和囊泡乙酰胆碱转运蛋白的定位”《化学神经解剖学杂志》。