title: Rapid detection of gene expression.
abstract: Conventional methods of in situ hybridization are not only complex, laborious and lengthy, tending to produce poor cellular resolution but are also inherently hazardous. We report a new method that overcomes many of these difficulties. Denovo attachment of a non-isotopic ligand, viz. 5-bromo-2'-deoxyuridine (BrdU) to oligonucleotide probes guaranteed the production of labelled probes of 100% specific activity that gave consistent, reproducible results, within hours, in a rapid in situ hybridization (ISH) technique. The method is demonstrated by localization of insulin-like growth factor II (IGF-II) mRNA in human intracranial tumours and foetal brain. Five of 10 meningiomas expressed IGF-II mRNA to an extent of 1% of their tumour cells and two of three pituitary adenomas exhibited a less than 0.2% IGF-II mRNA-positive cell population. IGF-II may be important in the progression and/or maintenance of these tumours. Foetal brain showed no hybridization. The method's simplicity, rapidity and reliability commends its use in a variety of research and diagnostic applications and the BrdU-labelled probes can be used in any hybridization technique.
authors.affiliation: Department of Neurosurgery, University of Birmingham, Warley, West Midlands, UK.
authors.forename: A
authors.initials: A
authors.lastname: Detta
authors.forename: E
authors.initials: E
authors.lastname: Hitchcock
mesh_terms: D000236:Adenoma / Q000378:metabolism; D001483:Base Sequence; D001921:Brain / Q000196:embryology / Q000378:metabolism; D001932:Brain Neoplasms / Q000378:metabolism; D001973:Bromodeoxyuridine; D015342:DNA Probes; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D007335:Insulin-Like Growth Factor II / Q000235:genetics* / Q000378:metabolism; D008579:Meningioma / Q000378:metabolism; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010911:Pituitary Neoplasms / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D012333:RNA, Messenger; D007335:Insulin-Like Growth Factor II; D001973:Bromodeoxyuridine