title: Enrichment of single neurons and defined brain regions from human brain tissue samples for subsequent proteome analysis.
abstract: 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.
authors.affiliation: Physiopathology in Aging Lab/Brazilian Aging Brain Study Group-LIM22, University of Sao Paulo Medical School, São Paulo, Brazil.
authors.forename: Mariana
authors.initials: M
authors.lastname: Molina
authors.forename: Simone
authors.initials: S
authors.lastname: Steinbach
authors.forename: Young Mok
authors.initials: YM
authors.lastname: Park
authors.forename: Su Yeong
authors.initials: SY
authors.lastname: Yun
authors.forename: Ana Tereza
authors.initials: AT
authors.lastname: Di Lorenzo Alho
authors.forename: Helmut
authors.initials: H
authors.lastname: Heinsen
authors.forename: Lea T
authors.initials: LT
authors.lastname: Grinberg
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
authors.forename: Renata E Paraizo
authors.initials: RE
authors.lastname: Leite
authors.forename: Caroline
authors.initials: C
authors.lastname: May
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001921:Brain / Q000166:cytology*; D005260:Female; D006801:Humans; D061346:Laser Capture Microdissection; D008297:Male; D008875:Middle Aged; D009474:Neurons / Q000378:metabolism*; D011180:Postmortem Changes; D020543:Proteome / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D020543:Proteome