Rapid in situ hybridization using digoxigenin probe and microwave oven.
In situ hybridization has been developed with probes labelled with a non-radioactive nucleotide, especially digoxigenin-coupled nucleotides. These non-radioactive probes significantly reduce safety problems and experimentation time. In this paper, we have studied by in situ hybridization the messenger RNA (mRNA) of the neuropeptide pro-opiomelanocortin (POMC), in the rat pituitary gland using digoxigenin labelled oligonucleotide and a microwave oven. Our protocol permitted us to visualize POMC mRNA in all cells of the intermediate lobe and a few corticotroph cells in the anterior lobe, as it has been already demonstrated and to complete the experiment in less than 24 hrs.
Besançon
R
R
INSERM U 433, Faculté de Médecine Alexis Carrel, Lyon, France.
Bencsik
A
A
Voutsinos
B
B
Belin
M F
MF
Févre-Montange
M
M
D000818:Animals; D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D004076:Digoxigenin; D005069:Evaluation Studies as Topic; D017403:In Situ Hybridization / Q000379:methods*; D008872:Microwaves; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000378:metabolism; D011333:Pro-Opiomelanocortin / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats
D015342:DNA Probes; D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin; D004076:Digoxigenin