title: UV-laser microdissection and mRNA expression analysis of individual neurons from postmortem Parkinson's disease brains.
abstract: Cell specificity of gene expression analysis is essential to avoid tissue sample related artifacts, in particular when the relative number of target cells present in the compared tissues varies dramatically, e.g., when comparing dopamine neurons in midbrain tissues from control subjects with those from Parkinson's disease (PD) cases. Here, we describe a detailed protocol that combines contact-free UV-laser microdissection and quantitative PCR of reverse-transcribed RNA of individual neurons from postmortem human midbrain tissue from PD patients and unaffected controls. Among expression changes in a variety of dopamine neuron marker, maintenance, and cell-metabolism genes, we found that α-synuclein mRNA levels were significantly elevated in individual neuromelanin-positive dopamine midbrain neurons from PD brains when compared to those from matched controls.
authors.affiliation: Wolfson Institute for Biomedical Research, University College London, London, UK.
authors.forename: Jan
authors.initials: J
authors.lastname: Gründemann
authors.forename: Falk
authors.initials: F
authors.lastname: Schlaudraff
authors.forename: Birgit
authors.initials: B
authors.lastname: Liss
mesh_terms: D001921:Brain / Q000378:metabolism / Q000473:pathology*; D002469:Cell Separation / Q000379:methods; D015925:Cryopreservation; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007834:Lasers; D008543:Melanins / Q000378:metabolism; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D008867:Microtomy / Q000379:methods; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D059010:Single-Cell Analysis / Q000379:methods*; D014466:Ultraviolet Rays
chemical_list: D008543:Melanins; D012333:RNA, Messenger; C014121:neuromelanin