title: Immunocytochemistry of 5-bromo-2'-deoxyuridine labelled oligonucleotide probes. A novel technique for in situ hybridization.
abstract: A synthetic oligonucleotide probe, complementary to oxytocin m-RNA was labelled enzymatically with 5-bromo-2'-deoxyuridine (5-BrdU) and with [gamma-32P]-ATP. The labelled probes were used for in situ hybridization of histological sections of the mouse hypothalamus. A monoclonal antibody to 5-BrdU and the streptavidine-peroxidase technique were used in order to visualize hybridization with the 5-BrdU labelled probe. In situ hybridization with [32P] labelling was detected autoradiographically. With both methods hybridized neurons were visible in the magnocellular hypothalamic nuclei. While immunostaining and radio-labelling provided similar localization of oxytocin m-RNA, only the immunocytochemical technique showed clear cellular resolution of the reaction product. In situ hybridization with 5-BrdU labelled probes followed by 5-brdU immunocytochemistry seems to be a powerful alternative to common autoradiographic techniques.
authors.affiliation: Abteilung Anatomie und Zellbiologie, Universität Ulm, Federal Republic of Germany.
authors.forename: G F
authors.initials: GF
authors.lastname: Jirikowski
authors.forename: J F
authors.initials: JF
authors.lastname: Ramalho-Ortigao
authors.forename: T
authors.initials: T
authors.lastname: Lindl
authors.forename: H
authors.initials: H
authors.lastname: Seliger
mesh_terms: D000255:Adenosine Triphosphate; D000818:Animals; D001973:Bromodeoxyuridine; D005260:Female; D007031:Hypothalamus / Q000378:metabolism*; D007150:Immunohistochemistry; D051379:Mice; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides; D010121:Oxytocin / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*
chemical_list: D009841:Oligonucleotides; D012333:RNA, Messenger; D010121:Oxytocin; D000255:Adenosine Triphosphate; D001973:Bromodeoxyuridine