Dynamics of postsynaptic density proteins and its relation to synapse functions

突触后密度蛋白的动态及其与突触功能的关系

基本信息

批准号：
15200027
负责人：
OKABE Shigeo
金额：
$ 28.29万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15200027/
关键词：
synapse glutamate receptors plasticity fluorescence microscopy cultured neurons 神経培養

项目摘要

Postsynaptic density is an electron dense structure beneath the postsynaptic membrane of glutamatergic synapses. To reveal molecular mechanisms of its assembly, we performed the following experiments.1. We developed a novel method of estimating absolute number of PSD scaffolding molecules (PSD-95, GKAP, Shank, Homer) per single synapses. Based on the fluorescence intensity of microspheres calibarated against single GFP molecules, the number of GFP-tagged PSD molecules in single synapses was determined in cultured hippocampal neurons. Single synapses contain 100-400 of PSD molecules and the combined molecular mass of the four PSD molecules corresponds to 10% of the total mass of the PSD.2. RNA-binding protein TLS changes its distribution from dendritic shafts to spines along the course of dendritic maturation. TLS also translocates from the shaft cytoplasm to the spines by stimulation of metabotropic glutamate receptors. TLS-null neurons show abnormal development of spine morphology. These results suggest regulation of spine morphology by TLS after its activity-dependent transport into spines.3. We established multiple lines of transgenic mice expressing GFP-tagged PSD proteins. Using dissociated culture of hippocampal neurons derived from these transgenic mice, morphological changes of single spines were traced for more than a week, illustrating the transition of immature filopodia-like protrusions into mature spine-like structures. Long-term observation also revealed coordinated changes of PSD density in multiple dendrites of single neurons, suggesting integrative mechanism of synapse growth/elimination across different dendritic segments.In summary, these results indicate continual remodeling of scaffolding proteins, which serve as a major framework of the PSD structure and also suggest an important role of protein synthesis from dendritic mRNA in the maintenance of glutamatergic synapses.

突触后密度是谷氨酸能突触的突触后膜下方的电子致密结构。为了揭示其组装的分子机制，我们进行了以下实验。1。我们开发了一种新的方法，用于估计单个突触的绝对数量的PSD支架分子（PSD-95，GKAP，Shank，Homer）。基于微球对单个GFP分子的荧光强度，在培养的海马神经元中确定了单个突触中GFP标记的PSD分子的数量。单个突触包含100-400的PSD分子，四个PSD分子的组合分子质量对应于PSD.2的总质量的10％。 RNA结合蛋白TLS在树突成熟过程中将其分布从树突轴变为棘突。 TL还通过刺激代谢型谷氨酸受体从轴细胞质转移到脊柱。 TLS无效神经元表现出脊柱形态的异常发育。这些结果表明，TLS的活性依赖性转运到棘中3。我们建立了表达GFP标记的PSD蛋白的多种转基因小鼠。使用源自这些转基因小鼠的海马神经元的解离培养，将单个棘的形态变化追溯了一周以上，这说明了未成熟的丝状突起过渡到成熟的脊柱样结构。长期观察还揭示了单个神经元多个树突中PSD密度的协调变化，表明了在不同树突段之间突触生长/消除的整合机制。总而言之，这些结果表明，这些结果表明脚手架蛋白的持续重塑，这是PSD结构的主要框架，也表明了蛋白质合成的重要框架，该蛋白质的作用是Dender systeric of dendenr systra的作用。突触。

项目成果

期刊论文数量（48）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

High-purity lineage selection of embryonic stem cell-derived neurons.

胚胎干细胞来源的神经元的高纯度谱系选择。

DOI：
发表时间：
2005
期刊：
Stem Cells and Development 14
影响因子：
0
作者：
Schmandt;T.;Meents;E.;Gossrau;G.;Gornik;V.;Okabe;S.;O.Brustle
通讯作者：
O.Brustle

Ebihara, T., Kawabata, I., Usui, S., Sobue, K., S.Okabe: "Synchronized formation and remodeling of postsynaptic densities : long-term visualization of hippocampal neurons expressing postsynaptic density proteins tagged with GFP"Journal of Neuroscience. 23

Ebihara, T.、Kawabata, I.、Usui, S.、Sobue, K.、S.Okabe：“突触后密度的同步形成和重塑：表达 GFP 标记的突触后密度蛋白的海马神经元的长期可视化”杂志

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

p250GAP, a novel brain-enriched GTPase-activating protein for Rho family GTPase, is involved in the NMDA receptor signaling.

p250GAP 是 Rho 家族 GTPase 的一种新型大脑富集 GTPase 激活蛋白，参与 NMDA 受体信号传导。

DOI：
发表时间：
2003
期刊：
Molecular Biology of the Cell 13
影响因子：
0
作者：
Nakazawa;T.;Watabe;A.M.;Tezuka;T.;Yoshida;Y.;Yokoyama;K.;Umemori;H.;Inoue;A.;Okabe;S.;Manabe;T.;T.Yamamoto.
通讯作者：
T.Yamamoto.

Synchronized formation and remodeling of postsynaptic densities : long-term visualization of hippocampal neurons expressing postsynaptic density proteins tagged with GFP.

突触后密度的同步形成和重塑：表达 GFP 标记的突触后密度蛋白的海马神经元的长期可视化。