Journal of Neuroscience, Vol 8, 3459-3469, Copyright © 1988 by Society for Neuroscience
FMRFamide peptides in Helisoma: identification and physiological actions at a peripheral synapse
AG Bulloch, DA Price, AD Murphy, TD Lee and HN Bowes
Neuroscience Research Group, University of Calgary, Alberta, Canada.
Previous reports have demonstrated powerful neuromodulatory actions of the
molluscan tetrapeptide FMRFamide in both the central and peripheral nervous
systems of the freshwater snail Helisoma. The present study was designed to
examine both the nature of the FMRFamide-like peptides in Helisoma and to
define their physiological actions at a peripheral synapse. We report that,
as determined by HPLC/RIA and mass spectrometry, Helisoma contains both
FMRFamide and 2 of its analogs, FLRFamide and GDPFLRFamide. Whereas whole
animals contain about 100 pmol/gm of these peptides, they were enriched in
the nervous system (3000 pmol/gm) and in a peripheral target organ, the
salivary glands (500 pmol/gm). For histochemical and physiological studies
we examined the salivary glands, which are known to be innervated by neuron
4 of the buccal ganglion. We confirmed the presence of FMRFamide-like
fibers on the salivary gland by immunohistochemistry using a polyclonal
antiserum. These fibers appear to be largely derived from somata located in
the central ring ganglia. For physiological tests we examined the neuron
4-gland synapse, at which presynaptic action potentials normally evoke a
suprathreshold EPSP in gland cells. Bath application of FMRFamide,
FLRFamide, or GDPFLRFamide at micromolar concentration to a buccal
ganglia/salivary gland preparation completely suppressed spontaneous
rhythmic activity. The sites of action of these peptides were examined by
iontophoretic application of FMRFamide to neuron 4 or the salivary gland.
Application of the peptide to the soma of neuron 4 caused a
hyperpolarization that suppressed spontaneously generated action
potentials. When applied to the salivary gland, FMRFamide caused a
hyperpolarization that reduced the EPSPs evoked by neuron 4 to below spike
threshold. The latter observation implies a postsynaptic locus of action
for FMRFamide, and this possibility was tested by direct depolarization of
the gland with iontophoresis of ACh (the putative transmitter of neuron 4).
Such depolarizations were also reduced by FMRFamide. We conclude that
Helisoma contains FMRFamide and 2 of its analogs, these peptides being
enriched in the nervous system and salivary glands. Furthermore, these
peptides can suppress activation of the salivary glands by actions both
directly on gland cells and on the effector neuron.