Vertebrate Memory Models: Alterations in Neuronal Phosphoproteins
The intent of this research is to demonstrate the role of synaptic phosphoproteins in neuronal plasticity. We believe this research represents the first demonstration of a long-term alteration in phospho-rylational state of a post-synaptic membrane protein as a consequence of neuronal activation. The model a long-term plasticity employed in this study is the kindling model (Goddard et al., 1969). Specifically, we have used the “rapid kindling” method whereby full motor convulsions can be elicited by stimulation of the hippocampus in 3–6 hrs. as opposed to 30–60 days in normal kindling. Rapid kindling displays all the classical characteristics of normal kindling (Zorumski and Hatlelid, 1982). Essentially, rats are stimulated in biphasic square wave at a frequency of 10 Hz. for 10 seconds. Five days after the kindling program, rats were decapited, their brains removed and the hippocampus-entorhinal cortex and frontal cortex dissected out. Synaptic membranes were isolated and then incubated with[γ-32p]ATP. The degree of phosphate incorporation into various protein bands was determined by one and two dimensional SDS Polyacrylamide gel electrophoresis. Synaptic glycoproteins were isolated by concanavilin A affinity chromatography and then separated by SDS Polyacrylamide gel electrophoresis.