The molluscan cardioactive neuropeptide FMRFamide: Subcellular localization in bivalve ganglia

Ganglia of the marine mollusk Macrocallista nimbosa were pooled, homogenized, and subjected to differential centrifugation. The neuropeptide Phe-Met-Arg-Phe-NH₂ (FMRFamide) was concentrated in the microsomal pellet. When the medium-speed supernatant was centrifuged in a discontinuous sucrose gradient, three separate peaks of activity were detected and identified as acetylcholine, 5-hydroxytryptamine, and FMRFamide. The relative concentration of FMRFamide in each fraction was determined by bioassay and by radioimmunoassay (RIA). Both determinations revealed a peak of peptide in the middle of the sucrose gradient. Electron micrographs of each of the gradient interfaces were analyzed. The interface containing the peak of biological FMRFamide activity was enriched two- to fivefold in neurosecretory granules with a mean diameter of 104 nm and various electron densities. Morphologically similar vesicles were also seen in intact ganglia. These findings support the notion that FMRFamide is a neurosecretory product. But the physiological function of the peptide in bivalve ganglia remains unknown.