Immunocytochemistry of 5-bromo-2?-deoxyuridine labelled oligonucleotide probes

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Abstract
A synthetic oligonucleotide probe, complementary to oxytocin m-RNA was labelled enzymatically with 5-bromo-2′-deoxyuridine (5-BrdU) and, with [γ-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.