NERVE GROWTH-FACTOR TREATMENT OF SENSORY NEURON PRIMARY CULTURES CAUSES ELEVATED LEVELS OF THE MESSENGER-RNA 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 induce d by nerve growth factor, within 60 min, in cultured adult rat dorsal root ganglion neurons. ATP synthase beta-subunit cDNA clones were isol ated from a lambda library. The library was constructed using rat dors al root ganglion mRNA that was differentially screened with cDNA-deriv ed probes from untreated and nerve-growth-factor-treated primary cultu res of adult rat dorsal root ganglion sensory neurons. Radiolabelled p robes were made from submicrogram quantities of RNA, by a novel PCR-ba sed technique, which allows small amounts of primary tissue to be used for library screening. The use of this technique in isolating novel d ifferentially expressed mRNAs is discussed.