The broad objectives of this research program are to identify specific proteins and biochemical processes that may be important for information processing in the CNS. So far, we have been able to demonstrate that the metabolism of three goldfish brain proteins (alpha, beta, and gamma) is markedly enhanced after learning. Two of these proteins (beta and gamma) were purified and shown to be glycoproteins with a 5 percent and 1.5 percent carbohydrate content. Immunohistofluorescence studies indicate that the proteins are localized in specific cells within the ependymal zone of goldfish brain. Beta and gamma appear to be rapidly synthesized and secreted into the extracellular fluid (ECF). Antisera to these """"""""secreted factors,"""""""" when injected into the brains of trained animals, can cause amnesia for recently acquired behavior. We have developed tissue culture methods for the ependymal cells of goldfish brain. Amino acid incorporation studies and radioimmunoassay methods will be used to define the pattern of synthesis and secretion of beta and gamma. Also, we will look for agents that can stimulate beta and gamma synthesis. In pilot studies, we find that proteins similar to beta and gamma are present in mammalian brain (mouse). These are rapidly labeled and secreted into ECF after the mice are trained in a T-maze. The pattern and time course of labeling of these proteins will be investigated as a function of training and for a variety of control experiments. The proteins will subsequently be isolated, purified, and characterized by immunochemical and biochemical methods. The multifaceted approach of this research program will help in developing an understanding of the biochemistry of learning and memory. The research should aid in the search for pharmacological agents which stimulate learning and improve the cognitive function of the mentally retarded, the chronically alcoholic, and the senile demented individuals.
