Neuronal sites of action of a neurosecretory peptide, egg-laying hormone, in Aplysia californica

Abstract

1. Egg-laying hormone (ELH) is a polypeptide of about 4,500 mol wt synthesized in the bag cell neurons of the abdominal ganglion of Aplysia. We studied the effects of ELH on the neuronal activity of the attached head ganglia (buccal, cerebral, pleural, and pedal), on the isolated buccal ganglia, as well as on feeding in intact Aplysia. 2. Starved animals (n = 7) injected with crude extract containing ELH stopped eating algae at 17 +/- 4 min and their eggs first appeared at 29 +/- 4 min after injection at 20 degrees C. This cessation of eating is significant when compared to the seven controls (P less than 0.01). These data clearly indicate that a suppression of feeding activity occurs before the appearance of eggs. 3. ELH applied to the paired buccal ganglia in vitro activates a pair of neurons into a tonic pacemaker mode (approximately 1 spike/s). This activation also occurs in a high-magnesium, zero calcium solution that blocks chemical synapses. The time for the full appearance of this activity in vitro correlates well with the time for suppression of feeding in vivo. Each of these neurons has an ipsilateral axon in buccal nerve 3. The neuron has been identified by intracellular recording. 4. ELH increases the rate of firing of a second pair of buccal neurons, each with an ipsilateral axon in the cerebrobuccal connective. 5. ELH, when applied to the attached head ganglia, causes large bursts of neuronal activity in pedal nerves to the foot and increased activity in the nerve to the penis; the relevant neurons remain to be identified. 6. These in vitro effects were produced by ELH partially purified from bag cell cluster homogenates using ammonium sulfate precipitation followed by anion exchange and gel filtration chromatography or by ELH released from activated bag cells in isolated abdominal ganglia and then purified by gel filtration. The isolated buccal ganglia effects have been confirmed with fully purified ELH. 7. The ELH effects on the in vitro nervous system support the hypothesis that ELH in vivo acts directly on the nervous system to suppress feeding activity, controlled by the buccal and cerebral ganglia. ELH may also produce characteristic movements of the head during egg laying, controlled probably by the pedal and cerebral ganglia.