Nerve growth factor treatment of sensory neuron primary cultures causes elevated levels of the mRNA encoding the ATP synthase beta-subunit as detected by a novel PCR-based differential cloning method.
The mRNA encoding the rat ATP synthase beta-subunit was rapidly induced by nerve growth factor, within 60 min, in cultured adult rat dorsal root ganglion neurons. ATP synthase beta-subunit cDNA clones were isolated from a lambda library. The library was constructed using rat dorsal root ganglion mRNA that was differentially screened with cDNA-derived probes from untreated and nerve-growth-factor-treated primary cultures of adult rat dorsal root ganglion sensory neurons. Radiolabelled probes were made from submicrogram quantities of RNA, by a novel PCR-based technique, which allows small amounts of primary tissue to be used for library screening. The use of this technique in isolating novel differentially expressed mRNAs is discussed.
Kendall
G
G
Department of Molecular Pathology, University College London Medical School, England.
Ensor
E
E
Crankson
H D
HD
Latchman
D S
DS
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers; D005727:Ganglia, Spinal / Q000201:enzymology*; D015870:Gene Expression; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000494:pharmacology*; D016133:Polymerase Chain Reaction / Q000379:methods; D006180:Proton-Translocating ATPases / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D013997:Time Factors
D017931:DNA Primers; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D006180:Proton-Translocating ATPases