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.
Pharmacology, Toxicology and Pharmaceutical Science
2
3%
Other
5
6%
Unknown
15
19%
Attention Score in Context
Attention Score in Context
This research output has an Altmetric Attention Score of 22. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 13 September 2022.
All research outputs
#1,725,973
of 25,769,258 outputs
Outputs from PLOS ONE
#21,126
of 224,419 outputs
Outputs of similar age
#4,394
of 98,417 outputs
Outputs of similar age from PLOS ONE
#67
of 524 outputs
Altmetric has tracked 25,769,258 research outputs across all sources so far. Compared to these this one has done particularly well and is in the 93rd percentile: it's in the top 10% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 224,419 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 15.8. This one has done particularly well, scoring higher than 90% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 98,417 tracked outputs that were published within six weeks on either side of this one in any source. This one has done particularly well, scoring higher than 95% of its contemporaries.
We're also able to compare this research output to 524 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 87% of its contemporaries.
