Regulatory Mechanism of Synthesis of Peptides and Receptors in Motoneurons : in Situ Hybridization Study

运动神经元肽和受体合成的调控机制：原位杂交研究

• 批准号: 01570027
• 负责人: SENBA Emiko
• 金额: $ 1.34万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570027/
• 关键词: Motoneuron; Axotomy; In Situ Hybridization; Galanin; CCK; CGRP; Neuronal Hicotinic Ach Receptor; NGF receptor; αーCGRP; βーCGRP; in situ ハイブリダイゼ-ション; NGF リセプタ-; axotomy(軸索切断); ペプタイド; NGFーリセプタ-; アセチルコリン・リセプタ-; mRNA

Acetylcholine (Ach) has been a sole candidate of neurotransmitter in motoneurons for a long time. Recent advance of molecular biology and applied morphological techniques have revealed the presence of neuropeptides in moto-neurons. These peptides are considered to modulate the activity of moto-neurons and muscles, or exert neurotrophic effects on them. In addition to peptides, neurotransmitter receptors are also synthesized in moto-neurons indicating that various kinds of neurotransmitters regulate the activity of motoneurons. Moreover, motoneurons are under the control of neurotrophic factors, such as nerve growth factor (NGF). It has been demonstrated that both NGF-receptor mRNA and NGF-receptor protein are expressed in developing motoneurons, and also in mature motoneurons under some experimental conditions. In the present study, we employed in situ hybridization histochemistry to demonstrate how the production of these peptides and receptors are regulated in injured motoーneurons.It … More is well documented that calcitonin gene-related peptide (CGRP)-like immunoreactivity in motoneurons of the rat spinal cord is increased after peripheral axotomy. In these neurons, axotomy caused the increase of alpha -CGRP mRNA, but not beta -CGRP mRNA, indicating that alpha -CGRP and beta -CGRP are regulated independently and have different roles in motoneurons. We have also shown that alpha - and beta -subunits constructing neuronal nicotinic Ach receptor are also differentially regulated in motoneurons. alpha 3 and beta 2 subunit mRNAs were expressed in 12% and 40% of facial motoneurons of the rat. alpha 3 subunit mRNA signals disappeared in response to axotomy, whereas the beta 2 subunit mRNA signal was remarkably enhanced.Some 38%, 55% and 7% of the facial motoneurons of the rat expressed alpha -CGRP, beta -CGRP and CCK mRNAs, respectively. No galanin mRNA containing motoneurons were observed. The levels of mRNA for alpha -CGRP, CCK and galanin were increased while beta -CGRP mRNA level was decreased after axotomy. The levels of mRNAs for these peptides returned to the control values by 2-4 weeks after injury, nerve transection had greater effects on beta -CGRP, CCK and galanin mRNAs than did nerve crush. These changing patterns of neuropeptides in motoneurons do not coincide with that of NGF-receptor mRNA after the same procedures ; the increase of NGF-receptor mRNA with crushing was more pronounced than with transection from the 3rd to the 14th day after the injury. These findings suggest that the production of these peptides in motoneurons is not regulated by NGF. Less

乙酰胆碱（Ach）是运动神经元中唯一的候选神经递质。近年来，分子生物学和形态学技术的发展揭示了运动神经元中神经肽的存在。这些肽被认为调节运动神经元和肌肉的活性，或对它们发挥神经营养作用。运动神经元除了合成肽类物质外，还合成神经递质受体，表明多种神经递质调节运动神经元的活动。此外，运动神经元受神经营养因子如神经生长因子（NGF）的控制。已经证明，在发育中的运动神经元中表达NGF受体mRNA和NGF受体蛋白，并且在某些实验条件下在成熟的运动神经元中也表达。在本研究中，我们采用原位杂交组织化学方法来证明这些肽和受体的产生是如何在受损的运动神经元中调节的。 ...更多信息 有充分的证据表明，大鼠脊髓运动神经元中降钙素基因相关肽（CGRP）样免疫反应性在外周轴突切断后增加。在这些神经元中，轴突切断引起α-CGRP mRNA的增加，但不引起β-CGRP mRNA的增加，表明α-CGRP和β-CGRP在运动神经元中是独立调节的，并且具有不同的作用。我们还表明，α-和β-亚单位构成的神经元烟碱乙酰胆碱受体也差异调节运动神经元。α 3和β 2亚单位mRNA在大鼠面运动神经元中的表达分别为12%和40%。α 3亚基mRNA信号消失，β 2亚基mRNA信号明显增强，分别有38%、55%和7%的面运动神经元表达α-CGRP、β-CGRP和CCK。未观察到含甘丙肽mRNA的运动神经元。切断神经后，α-CGRP、CCK和甘丙肽的mRNA水平升高，β-CGRP的mRNA水平降低.损伤后2-4周，这些肽的mRNA水平恢复到对照值，神经切断对β-CGRP、CCK和甘丙肽mRNA的影响大于神经挤压。这些变化模式的运动神经元中的神经肽不符合相同的程序后，NGF受体mRNA的增加与破碎比横断伤后第3至第14天更明显。这些发现表明，这些肽在运动神经元的生产是不受神经生长因子。少

项目成果

K.Noguchi,E.Senba,Y.Morita,M.Sato and M.Tohyama: "αーCGRP and βーCGRP mRNAs are differentially regulated in the rat spinal cord and dorsal root ganglion." Molecular Brain Research. 7. 299-304 (1990)

K. Noguchi、E. Senba、Y. Morita、M. Sato 和 M. Tohyama：“α-CGRP 和 β-CGRP mRNA 在大鼠脊髓和背根神经节中受到不同程度的调节。” 7. 299 -304 (1990)

T. Saika, E. Senba, K. Noguchi, M. Sato, Y. Yoshida, T. Kubo, T. Matsunaga and M. Tohyama: "Effects of Nerve Crush and Transection on mRNA Levels for Nerve Growth Factor Receptor in the Rat Facial Motoneurons." Mol. Brain Res.9. 157-160 (1990)

T. Saika、E. Senba、K. Noguchi、M. Sato、Y. Yoshida、T. Kubo、T. Matsunaga 和 M. Tohyama：“神经挤压和横断对大鼠神经生长因子受体 mRNA 水平的影响

T.Saika,E.Senba et al.: "PreproCCK and preprogalanin mRNAs are upregulated in axotomizedd facial motoneurons" sumitted to a journal.

T.Saika、E.Senba 等人：“在轴突切除的面部运动神经元中，PreproCCK 和前甘丙肽 mRNA 上调”提交给期刊。

K. Noguchi, E. Senba, Y. Morita, M. Sato and M. Tohyama: "Alpha-CGRP and beta-CGRP mRNAs are differentially regulated in the rat spinal cord and dorsal root ganglion." Mol. Brain Res.7. 299-304 (1990)

K. Noguchi、E. Senba、Y. Morita、M. Sato 和 M. Tohyama：“α-CGRP 和 β-CGRP mRNA 在大鼠脊髓和背根神经节中受到差异性调节。”

仙波 恵美子: "ニュ-ロンの再生過程における分子生物学ー顔面神経核運動ニュ-ロンをモデルとしてー" 医学のあゆみ. 155. 5 (1990)

Emiko Senba：“神经元再生过程的分子生物学 - 使用面神经核运动神经元作为模型”医学史 155. 5 (1990)。