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Enrichment of single neurons and defined brain regions from human brain tissue samples for subsequent proteome analysis.

Journal of neural transmission (Vienna, Austria : 1996) (2015-07-01)
Mariana Molina, Simone Steinbach, Young Mok Park, Su Yeong Yun, Ana Tereza Di Lorenzo Alho, Helmut Heinsen, Lea T Grinberg, Katrin Marcus, Renata E Paraizo Leite, Caroline May
RESUMO

Brain function in normal aging and neurological diseases has long been a subject of interest. With current technology, it is possible to go beyond descriptive analyses to characterize brain cell populations at the molecular level. However, the brain comprises over 100 billion highly specialized cells, and it is a challenge to discriminate different cell groups for analyses. Isolating intact neurons is not feasible with traditional methods, such as tissue homogenization techniques. The advent of laser microdissection techniques promises to overcome previous limitations in the isolation of specific cells. Here, we provide a detailed protocol for isolating and analyzing neurons from postmortem human brain tissue samples. We describe a workflow for successfully freezing, sectioning and staining tissue for laser microdissection. This protocol was validated by mass spectrometric analysis. Isolated neurons can also be employed for western blotting or PCR. This protocol will enable further examinations of brain cell-specific molecular pathways and aid in elucidating distinct brain functions.

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