title: Increase in protein and tubulin mRNA synthesis in frog sensory neurons treated with the adenylate cyclase activator, forskolin.
abstract: We are interested in the role cyclic AMP may play as a mediator of growth factor-induced gene expression in regenerating peripheral nerves. As a first step, we investigated mRNA levels and protein synthesis in intact frog dorsal root ganglia (DRG) treated with the adenylate cyclase activator, forskolin in vitro. Forskolin (10-7 M) increased intraganglionic cyclic AMP concentration 3-fold within 40 min of application and maintained this concentration through 60 min, when the drug was withdrawn. Addition of forskolin to isolated DRG neurons for 1 h increased the incorporation of [3H]leucine into TCA-insoluble material beginning 12 h after the withdrawal of forskolin. Axonally transported labeled material was increased almost 2-fold by 12 h. The effect of forskolin could be blocked by the simultaneous addition of actinomycin D, but not if actinomycin D was added 1 h later. Northern and dot-blot analysis of RNA extracted from the treated ganglia indicated that an mRNA coding for an α-subunit of tubulin was increased by treatment with forskolin. 2D PAGE also demonstrated an increase in an α-subunit of tubulin. An increase in neuronal cyclic AMP appears to selectively increase the production of specific proteins and may contribute to the production of macromolecules involved in the initiation and stimulation of axonal regeneration.
authors.affiliation: Department of Human Physiology, University of California School of Medicine, Davis, CA 95616 (U.S.A.).
authors.forename: R C
authors.initials: RC
authors.lastname: Carlsen
authors.forename: M
authors.initials: M
authors.lastname: De Leon
authors.forename: W
authors.initials: W
authors.lastname: Tetzlaff
authors.forename: I M
authors.initials: IM
authors.lastname: Parhad
authors.forename: M A
authors.initials: MA
authors.lastname: Bisby