Study of neurofilament phosphorylation in long-term potentiation of hlppocampus

神经丝磷酸化在海马长时程增强中的研究

• 批准号: 11670942
• 负责人: NAKAMURA Yu
• 金额: $ 2.11万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670942/
• 关键词: neurofilament; phosphorylation; PKA; PKC; Alzheimer; phoshorylation; Rh0

Long-term potentiation (LTP) is a cellular model of neuronal plasticity that is thought to involve the phosphorylation of various proteins. NF-L is considered a possible substrate for phosphorylation. First, we prepared site-specific phosphorylation-dependent antibodies (abNFL26) against the phosphorylation site Ser-26 of Rho-kinase. The in vitro specificity of those antibodies were appreciated by dot and western blot analyses. During LTP stimulation of mouse hippocampal slices, the series of the site-specific phosphorylation-dependent antibodies against the phosphorylation sites, Ser-26, -51, -55, -57, demonstrates the increase in the phosphorylation level of Ser57 in NF-L, and the visualization of the localized distribution of Ser57 phosphorylation in a subpopulation of apical dendrites of the pyramidal neurons. Furthermore, Ser57 phosphorylation during LTP is suggested to be mediated by CaMKII.We show that NF-L is phosphorylated by CaMKII in a subpopulation of the apical dendrites during LTP, indicating that Ser57 is a novel phosphorylation site of NF-L in vivo related to the neuronal signal transduction. And these data implies that the phosphorylation of NF-L by CaMKII might be influenced, resulting in memory dysfuctions.

长时程增强(LTP)是一种神经元可塑性的细胞模型，被认为涉及各种蛋白质的磷酸化。核因子-L被认为是一种可能的磷酸化底物。首先，我们制备了针对Rho-Kinase的磷酸化位点Ser-26的位点特异性磷酸化依赖抗体(AbNFL26)。通过斑点分析和免疫印迹分析，评价这些抗体的体外特异性。在LTP刺激小鼠海马片的过程中，一系列针对磷酸化位点Ser26、Ser-51、Ser-55、Ser-57的特异性磷酸化依赖抗体证实了NF-L中Ser57的磷酸化水平增加，并显示了Ser57磷酸化在锥体神经元顶树突亚群中的定位分布。此外，LTP过程中Ser57的磷酸化可能是由CaMKII介导的。我们发现，在LTP过程中，顶端树突的一个亚群中有一种CaMKII对核因子-L的磷酸化，这表明Ser57是体内与神经元信号转导相关的一个新的核因子-L的磷酸化位点。提示CaMKII对核因子-L的磷酸化可能有影响，导致记忆功能障碍。

项目成果

Nakamura Y.,Hashimoto R.et al.: "Major phosphorylation site (Ser-55) of neurofilament L by cyclic AMP dependent protein kinase A in rat primary neuronal culture"J. Neurochem.. 74. 949-959 (2000)

Nakamura Y.，Hashimoto R.等人：“大鼠原代神经元培养物中环AMP依赖性蛋白激酶A对神经丝L的主要磷酸化位点（Ser-55）”J。

Nakamura,Y.,Hashimoto,R., et al.: "Major phosphorylation site (Ser-55) of neurofilament L by cyclic AMP dependent protein kinase A in rat primary neuronal culture."J.Neurochem.. 74. 949-959 (2000)

Nakamura,Y.,Hashimoto,R., et al.：“大鼠原代神经元培养物中环 AMP 依赖性蛋白激酶 A 对神经丝 L 的主要磷酸化位点 (Ser-55)。”J.Neurochem.. 74. 949-959

Hashimoto,R.,Nakamura,Y., et al.: "Site-specific phosphorylation of neurofilament-L is mediated by calcium/calmodulin dependent protein kinase II in the apical dendrites during long-term potentiation,"J.Neurochem.. 75. 373-382 (2000)

Hashimoto, R., Nakamura, Y. 等人：“在长期增强过程中，神经丝 L 的位点特异性磷酸化是由顶端树突中的钙/钙调蛋白依赖性蛋白激酶 II 介导的”，J.Neurochem.. 75

Hashimoto, R., Nakamura, Y., et al.: "Phosphorylation of neurofilament L during LTD"NeuroReport. 11. 739-2742 (2000)

Hashimoto, R.、Nakamura, Y. 等人：“LTD 期间神经丝 L 的磷酸化”NeuroReport。

Hashimoto,R.,Nakamura,Y., et al.: "Phosphorylation of neurofilament L during LTD"Neuro Report. 11. 739-2742 (2000)

Hashimoto,R.、Nakamura,Y. 等人：“LTD 期间神经丝 L 的磷酸化”神经报告。