| Title |
The Post-Synaptic Density of Human Postmortem Brain Tissues: An Experimental Study Paradigm for Neuropsychiatric Illnesses
|
|---|---|
| Published in |
PLOS ONE, April 2009
|
| DOI | 10.1371/journal.pone.0005251 |
| Pubmed ID | |
| Authors |
Chang-Gyu Hahn, Anamika Banerjee, Matthew L. MacDonald, Dan-Sung Cho, Joshua Kamins, Zhiping Nie, Karin E. Borgmann-Winter, Tilo Grosser, Angel Pizarro, Eugene Ciccimaro, Steven E. Arnold, Hoau-Yan Wang, Ian A. Blair |
| Abstract |
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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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 3 | 33% |
| Austria | 1 | 11% |
| United Kingdom | 1 | 11% |
| Unknown | 4 | 44% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 8 | 89% |
| Practitioners (doctors, other healthcare professionals) | 1 | 11% |
Mendeley readers
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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 2 | 3% |
| United States | 2 | 3% |
| Unknown | 73 | 95% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 25 | 32% |
| Student > Ph. D. Student | 15 | 19% |
| Student > Master | 7 | 9% |
| Student > Postgraduate | 4 | 5% |
| Professor | 4 | 5% |
| Other | 10 | 13% |
| Unknown | 12 | 16% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 31 | 40% |
| Neuroscience | 13 | 17% |
| Biochemistry, Genetics and Molecular Biology | 9 | 12% |
| Medicine and Dentistry | 3 | 4% |
| Pharmacology, Toxicology and Pharmaceutical Science | 2 | 3% |
| Other | 5 | 6% |
| Unknown | 14 | 18% |