The post-synaptic density of human postmortem brain tissues: an experimental study paradigm for neuropsychiatric illnesses.
The post-synaptic density of human postmortem brain tissues: an experimental study paradigm for neuropsychiatric illnesses.
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DOI：
10.1371/journal.pone.0005251
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Blair IA
中科院分区：
文献类型：
--
作者：
Hahn CG;Banerjee A;Macdonald ML;Cho DS;Kamins J;Nie Z;Borgmann-Winter KE;Grosser T;Pizarro A;Ciccimaro E;Arnold SE;Wang HY;Blair IA
最近的分子遗传学研究表明，各种跨突触过程的神经精神疾病的病理生理机制。对患者大脑中突触前和突触后支架的检查将极大地有助于进一步的研究，但这种方法在人类死后组织中还有待测试。我们已经检查了三种方法，使用基于密度梯度的突触体纯化，然后进行去污剂提取（方法1）和基于pH的突触膜差异提取（方法2和3）。所有这三种方法都从人类死后大脑中分离出高度富集典型PSD蛋白的组分，几乎排除了突触前蛋白。我们使用电子显微镜（EM）检查这些组分，并验证了来自人死后脑组织的突触膜和PSD组分的完整性。我们使用二维液相色谱串联质谱（2D LC-MS/MS）分析PSD组分的蛋白质组成，并通过质谱观察已知的PSD蛋白。免疫沉淀和免疫印迹研究表明，预期的蛋白质-蛋白质相互作用和某些转录后调节PSD组分中保持。我们的研究结果表明，PSD组分可以从人死后脑组织中分离出具有合理程度的完整性。这种方法可能会促进新的死后大脑研究范式，其中特定微区的化学计量和蛋白质组成进行检查。
Recent molecular genetics studies have suggested various trans-synaptic processes for pathophysiologic mechanisms of neuropsychiatric illnesses. Examination of pre- and post-synaptic scaffolds in the brains of patients would greatly aid further investigation, yet such an approach in human postmortem tissue has yet to be tested. We have examined three methods using density gradient based purification of synaptosomes followed by detergent extraction (Method 1) and the pH based differential extraction of synaptic membranes (Methods 2 and 3). All three methods separated fractions from human postmortem brains that were highly enriched in typical PSD proteins, almost to the exclusion of pre-synaptic proteins. We examined these fractions using electron microscopy (EM) and verified the integrity of the synaptic membrane and PSD fractions derived from human postmortem brain tissues. We analyzed protein composition of the PSD fractions using two dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS) and observed known PSD proteins by mass spectrometry. Immunoprecipitation and immunoblot studies revealed that expected protein-protein interactions and certain posttranscriptional modulations were maintained in PSD fractions. Our results demonstrate that PSD fractions can be isolated from human postmortem brain tissues with a reasonable degree of integrity. This approach may foster novel postmortem brain research paradigms in which the stoichiometry and protein composition of specific microdomains are examined.
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影响因子：
3.4
作者：
Crecelius, Anna;Goetz, Andrea;Kretzschmar, Hans A.
通讯作者：
Kretzschmar, Hans A.
影响因子：
4.7
作者：
Hunsucker, Stephen W.;Solomon, Benjamin;Patterson, David
通讯作者：
Patterson, David
不同大脑区域突触后密度的分离和表征：丰富不同类型的突触后密度。
DOI：
10.1083/jcb.86.3.831
发表时间：
1980-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Carlin RK;Grab DJ;Cohen RS;Siekevitz P
通讯作者：
Siekevitz P
影响因子：
7
作者：
Cheng, Dongmei;Hoogenraad, Casper C.;Peng, Junmin
通讯作者：
Peng, Junmin
DOI：
10.1073/pnas.0505359102
发表时间：
2005-08-09
影响因子：
11.1
作者：
Chen, XB;Vinade, L;Reese, TS
通讯作者：
Reese, TS