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PhD candidate Vasin Dumrongprechachan captured the protein expression in the mouse brain and analyzed it by mass spectrometry using a fluorescence microscope
.
The image shows a sagittal cross-section of the rat striatum
Researchers have developed a successful method for the first time to identify proteins in different types of neurons in the brains of living animals
.
The new research, led by Northwestern University and the University of Pittsburgh, is a big step towards understanding the millions of different proteins in the brain
.
As a component of all cells, including neurons, protein is the key to a better understanding of complex brain diseases, such as Parkinson’s and Alzheimer’s, which may lead to the development of new treatments
The research will be published in the journal Nature Communications on August 11
.
In this new study, the researchers designed a virus that can send an enzyme to a precise location in the brain of a living mouse
.
This enzyme extracted from soybeans genetically marks adjacent proteins in predetermined positions
"Similar work has been done in cell culture before
.
But the cells in the petri dish do not work like the cells in the brain.
By chemically labeling proteins and their neighbors, researchers can now understand how proteins work in specific controlled areas and how they interact in the proteome
.
In addition to the virus that carries the soybean enzyme, the researchers also used the virus to carry a separate green fluorescent protein
Kozorovitskiy said: "This virus is essentially a message we send
.
In this case, the message carries this special soybean enzyme
Protein targeting plays a catching-up role
Although gene targeting has completely changed biology and neuroscience, protein targeting has unfortunately lagged behind
.
Researchers can amplify and sequence genes and RNA to determine their exact components
Kozorovitskiy said: "We have been able to make great progress in gene and RNA sequencing, but proteins have not yet been involved
.
However, everyone recognizes the importance
"Proteomics based on mass spectrometry is a powerful technique," said Vasin Dumrongprechachan, a doctoral candidate in Kozorovitskiy's laboratory and the first author of the paper
.
"With our method, we can begin to map the proteome of various brain circuits with high precision and specificity
.
We can even quantify them to see how many proteins are present in neurons and different parts of the brain
.
"
Next step: better understanding of brain diseases
Now that this new system has been verified and ready, researchers can apply it to mouse models of diseases to better understand neurological diseases
.
Dumrongprechachan said: "We hope to extend this approach and begin to identify the biochemical modifications of neuronal proteins that occur in specific patterns of brain activity or changes caused by neuroactive drugs to promote clinical progress
.
"
Kozorovitskiy said: "We look forward to applying this research results to models related to brain diseases and linking these studies to the postmortem proteomics studies of the human brain
.
It is ready to be applied to these models, and we can’t wait.
Up
.
"
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Cell-type and subcellular compartment-specific APEX2 proximity labeling reveals activity-dependent nuclear proteome dynamics in the striatum.
10.
1038/s41467-021-25144-y
