In situ transcription: specific synthesis of complementary DNA in fixed tissue sections.
A technique, in situ transcription, is described, in which reverse transcription of mRNAs is achieved within fixed tissue sections. An oligonucleotide complementary to proopiomelanocortin (POMC) mRNA was used as a primer for the specific synthesis of radiolabeled POMC cDNA in fixed sections of rat pituitary, thus permitting the rapid anatomical localization of POMC mRNA by autoradiography. Intermediate lobe signal intensities were sensitive to dopaminergic drugs, demonstrating that the method can be used for studies of mRNA regulation. The transcripts may also be eluted from tissue sections for a variety of uses, including the identification and cloning of autoradiographically localized cDNAs from small amounts of tissue.
Tecott
L H
LH
Nancy Pritzker Laboratory of Behavioral Neurochemistry, Department of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, CA 94305.
Barchas
J D
JD
Eberwine
J H
JH
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis*; D003841:Deoxycytidine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics; D010902:Pituitary Gland / Q000378:metabolism*; D011333:Pro-Opiomelanocortin / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D051381:Rats; D014158:Transcription, Genetic
D009841:Oligonucleotides; D012333:RNA, Messenger; D003841:Deoxycytidine; D011333:Pro-Opiomelanocortin; D004247:DNA; D012194:RNA-Directed DNA Polymerase