title: RAKE and LNA-ISH reveal microRNA expression and localization in archival human brain.
abstract: microRNAs (miRNAs) are small (approximately 22 nucleotide) regulatory RNAs which play fundamental roles in many biological processes. Recent studies have shown that the expression of many miRNAs is altered in various human tumors and some miRNAs may function as oncogenes or tumor suppressor genes. However, with the exception of glioblastoma multiforme, the expression of miRNAs in brain tumors is unknown. Furthermore, methods to profile miRNAs from formalin-fixed, paraffin-embedded (FFPE) archival tissues or to study their cellular and subcellular localization in FFPE tissues have been lacking. Here we report the coordinated miRNA expression analysis from the tissue level to the subcellular level, using the RAKE (RNA-primed, array-based, Klenow Enzyme) miRNA microarray platform in conjunction with Locked Nucleic Acid (LNA)-based in situ hybridization (LNA-ISH) on archival FFPE human brains and oligodendroglial tumors. The ability to profile miRNAs from archival tissues at the tissue level, by RAKE microarrays, and at the cellular level by LNA-ISH, will accelerate studies of miRNAs in human diseases.
authors.affiliation: Division of Neuropathology and Department of Pathology & Laboratory Medicine, University of Pennsylvania School of Medicine, 605A Stellar-Chance Bldg., 421 Curie Blvd., Philadelphia, PA 19104, USA.
authors.forename: Peter T
authors.initials: PT
authors.lastname: Nelson
authors.forename: Don A
authors.initials: DA
authors.lastname: Baldwin
authors.forename: Wigard P
authors.initials: WP
authors.lastname: Kloosterman
authors.forename: Sakari
authors.initials: S
authors.lastname: Kauppinen
authors.forename: Ronald H A
authors.initials: RH
authors.lastname: Plasterk
authors.forename: Zissimos
authors.initials: Z
authors.lastname: Mourelatos
mesh_terms: D001921:Brain / Q000473:pathology / Q000502:physiology*; D001932:Brain Neoplasms / Q000235:genetics* / Q000473:pathology; D002878:Chromosomes, Human, Pair 1; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D035683:MicroRNAs / Q000032:analysis*; D046228:Microarray Analysis / Q000379:methods*; D009837:Oligodendroglioma / Q000235:genetics* / Q000473:pathology; D014015:Tissue Banks
chemical_list: D035683:MicroRNAs