Polypeptide Secretion from the Isolated Parietovisceral Ganglion of Aplysia californica

Abstract

In vitro studies of the secretory behavior of the parietovisceral ganglion in Aplysia californica were performed. The aim of these studies was to investigate the release of polypeptides in response to depolarizing stimuli, and, in particular, to determine if a specific polypeptide known to induce egg laying in the intact animal is secreted into the bathing medium. During continuous perfusion of a ganglion preincubated in leucine-³H the application of either high-potassium medium or a burst of electrical stimuli (via the pleurovisceral connective nerve) evoked a marked increase in the amount of trichloroacetic acid (TCA)-precipitable radioactivity recovered in the perfusate. Enhanced release could be detected within 80 sec of the initial exposure to high potassium; however, incubation of a ganglion in calcium-free media before the application of high-potassium medium abolished the increase of precipitable radioactivity. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of perfusate samples revealed a significant change in the polypeptide species washed from the ganglion during high-potassium depolarization. Bioassays confirmed that egg laying is induced when high-potassium medium used to bathe a ganglion is injected into a recipient animal. These and other results permit the conclusion that the bulk of the polypeptide material secreted from the ganglion in response to depolarization is a specific neurohormone produced by two identified cell clusters, the so-called bag cells.