title: A translational profiling approach for the molecular characterization of CNS cell types.
abstract: The cellular heterogeneity of the brain confounds efforts to elucidate the biological properties of distinct neuronal populations. Using bacterial artificial chromosome (BAC) transgenic mice that express EGFP-tagged ribosomal protein L10a in defined cell populations, we have developed a methodology for affinity purification of polysomal mRNAs from genetically defined cell populations in the brain. The utility of this approach is illustrated by the comparative analysis of four types of neurons, revealing hundreds of genes that distinguish these four cell populations. We find that even two morphologically indistinguishable, intermixed subclasses of medium spiny neurons display vastly different translational profiles and present examples of the physiological significance of such differences. This genetically targeted translating ribosome affinity purification (TRAP) methodology is a generalizable method useful for the identification of molecular changes in any genetically defined cell type in response to genetic alterations, disease, or pharmacological perturbations.
authors.affiliation: Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Myriam
authors.initials: M
authors.lastname: Heiman
authors.forename: Anne
authors.initials: A
authors.lastname: Schaefer
authors.forename: Shiaoching
authors.initials: S
authors.lastname: Gong
authors.forename: Jayms D
authors.initials: JD
authors.lastname: Peterson
authors.forename: Michelle
authors.initials: M
authors.lastname: Day
authors.forename: Keri E
authors.initials: KE
authors.lastname: Ramsey
authors.forename: Mayte
authors.initials: M
authors.lastname: Suárez-Fariñas
authors.forename: Cordelia
authors.initials: C
authors.lastname: Schwarz
authors.forename: Dietrich A
authors.initials: DA
authors.lastname: Stephan
authors.forename: D James
authors.initials: DJ
authors.lastname: Surmeier
authors.forename: Paul
authors.initials: P
authors.lastname: Greengard
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Heintz
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D002490:Central Nervous System / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial / Q000378:metabolism; D003042:Cocaine / Q000494:pharmacology; D018765:Dopamine Uptake Inhibitors / Q000494:pharmacology; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D008822:Mice, Transgenic; D008954:Models, Biological; D009474:Neurons / Q000378:metabolism; D014176:Protein Biosynthesis; D012270:Ribosomes / Q000378:metabolism
chemical_list: D018765:Dopamine Uptake Inhibitors; D049452:Green Fluorescent Proteins; D003042:Cocaine