Brain protein metabolism and the acquisition of new behaviors. II. Immunological studies of the alpha, beta and gamma proteins of goldfish brain.

In a previous study, the labeling pattern of three proteins (alpha, beta and gamma) in goldfish brain was found to change after the animals successfully acquired a new pattern of behavior. In the present study, these proteins were isolated from the brain cytoplasmic fraction, purified by successive gel electrophoresis and used as antigent to immunize rabbits. Antisera containing antibodies to two of the proteins (beta and gamma) were obtained. These gave single precipitin bands when plated against the antigens and a mixture of the total cytoplasmic proteins. The distribution of beta and gamma in brain subcellular fractions and in a variety of goldfish tissues was determined by immunodiffusion methods. gamma was specific to brain. The beta protein cross-reacted but was not identical to a widely distributed substance in plasma, liver and kidney. Both beta and gamma appear to be species specific in that no cross-reactivity was obtained with mouse, chick or rat brain proteins. Immunological methods, in combination with double labeling experiments were used to establish that the beta and gamma antigens were proteins which were normally present in goldfish brain. Both the beta and gamma antisera were equally capable of specifically precipitating the proteins which were differentially labeled after training as well as purified proteins of the same molecular weight present in the brains of control animals. These results suggest that the acquisition of a new pattern of behavior can increase the demand for the synthesis of specific proteins (beta and gamma) normally present in goldfish brain.