title: Protein analysis through Western blot of cells excised individually from human brain and muscle tissue.
abstract: Comparing protein levels from single cells in tissue has not been achieved through Western blot. Laser capture microdissection allows for the ability to excise single cells from sectioned tissue and compile an aggregate of cells in lysis buffer. In this study we analyzed proteins from cells excised individually from brain and muscle tissue through Western blot. After we excised individual neurons from the substantia nigra of the brain, the accumulated surface area of the individual cells was 120,000, 24,000, 360,000, 480,000, 600,000 μm2. We used an optimized Western blot protocol to probe for tyrosine hydroxylase in this cell pool. We also took 360,000 μm2 of astrocytes (1700 cells) and analyzed the specificity of the method. In muscle we were able to analyze the proteins of the five complexes of the electron transport chain through Western blot from 200 human cells. With this method, we demonstrate the ability to compare cell-specific protein levels in the brain and muscle and describe for the first time how to visualize proteins through Western blot from cells captured individually.
authors.affiliation: Department of Neurology, Klinikum Grosshadern, University of Munich, Marchinionistr. 23, 81377 Munich, Germany. andrew.koob@uwrf.edu
authors.forename: A O
authors.initials: AO
authors.lastname: Koob
authors.forename: L
authors.initials: L
authors.lastname: Bruns
authors.forename: C
authors.initials: C
authors.lastname: Prassler
authors.forename: E
authors.initials: E
authors.lastname: Masliah
authors.forename: T
authors.initials: T
authors.lastname: Klopstock
authors.forename: A
authors.initials: A
authors.lastname: Bender
mesh_terms: D001253:Astrocytes / Q000378:metabolism; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D003576:Electron Transport Complex IV / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D061346:Laser Capture Microdissection; D009132:Muscles / Q000378:metabolism* / Q000473:pathology; D009474:Neurons / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis*
chemical_list: D014446:Tyrosine 3-Monooxygenase; D003576:Electron Transport Complex IV