Immunocytochemistry of 5-bromo-2'-deoxyuridine labelled oligonucleotide probes. A novel technique for in situ hybridization.
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.
Jirikowski
G F
GF
Abteilung Anatomie und Zellbiologie, Universität Ulm, Federal Republic of Germany.
Ramalho-Ortigao
J F
JF
Lindl
T
T
Seliger
H
H
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*
D009841:Oligonucleotides; D012333:RNA, Messenger; D010121:Oxytocin; D000255:Adenosine Triphosphate; D001973:Bromodeoxyuridine