We have developed a rapid sensitive test for factors that mimic the trophic effects of nerves by maintaining normal rates of protein synthesis in denervated forelimb blastemata of adult newts (Notophthalmus viridescens). Rates of protein synthesis in secondary blastemata are similar. However, after they are denervated and explanted into organ culture the rates of protein synthesis first increase and later fall below control values. Similar changes occur after denervation in vivo. The alterations in the rates of protein synthesis are prevented by adding to the culture medium aqueous extracts from brains of adult newts or chicken embryos. The active material is either a peptide or a protein.

No abstract

Adult newts regenerate functional limbs after amputation. This process normally depends on the trophic influence of nerves on the regenerating limbs, particularly in the early stages before differentiation of the regeneration blastema, when it stimulates growth by maintaining high rates of macromolecular synthesis. The sequence of biochemical events involved is unknown, but it has been suggested that intracellular cyclic AMP may be a second messenger within the blastema. Many studies have indicated that the neural agent(s) involved might be protein. The recent finding that blastemata contain high levels of catecholamines, however, has implicated noradrenaline (NA) as the neurotrophic agent, and suggested that it works via stimulation of beta-adrenergic receptors on the blastemal cells, thereby raising the intracellular concentrations of cyclic AMP. To test this hypothesis we studied the ability of NA alone and in combination with alpha-and beta-adrenergic antagonists to increase cyclic AMP levels and to mimic the effects of nerves by maintaining high rates of protein synthesis and high mitotic indices (MI) in denervated blastemata in organ culture. We find that although NA raises cyclic AMP levels through a beta-adrenergic effect, it does not maintain high rates of protein synthesis or high MI in cultured blastemata. It is unlikely therefore, that this hypothesis applies.

Three experiments were performed to assess the interanimal transferability of conditioned taste aversion to 0.1% saccharin. Two experiments used an intracerebrospinal fluid (subdural) route for administering brain extracts and a third used an intraperitoneal (IP) route. As assessed by repeated measurements ANOVA, saccharin consumption was significantly lower during extinction of conditioned aversion for experimental recipients (ER) receiving extracts from aversively conditioned donors, than that of control recipients (CR), receiving extracts from unconditioned donors in one subdural experiment, F(21, 189) = 1.61, p less than 0.05. In the IP experiment the results were in the same direction, though not significant, F(34, 238) = 1.39, p less than 0.1. Results of the other subdural experiment are discussed. It is concluded that these experiments with the conditioned taste aversion paradigm have potential as a model for investigations of behavioral interanimal transfer (BIT) and for neuromolecular research aimed at identification of associated putative neurochemical(s) and the elaboration of their mechanism of action.