Localization and immunological characterization of insulin-like peptide(s) in the land snail Otala lactea (Mollusca: Pulmonata)

Abstract

Insulin-like peptides were detected by means of immunological techniques in tissues of the land snail Otala lactea. Insulin-positive cells were detected in all the ganglia except the right parietal ganglion and visceral ganglion. In the digestive tract, insulin-positive cells were found in the muscle and connective tissue layer of the intestine. The amount of insulin-like peptide detected in acid-ethanol extract of brains and digestive tracts from active snails did not differ significantly from that in the corresponding tissues from estivating (dormant) ones. More insulin-like peptide was detected in hemolymph from active snails than in hemolymph from estivating ones. Brains from active snails released insulin-like peptide in vitro. Analysis of the cerebral ganglia or digestive tract extracts by size-exclusion chromatography and insulin RIA revealed more than one fraction with insulin immunoreactivity. Some of these fractions contained material with molecular masses close to those of mammalian insulin or its subunits. Further analysis of the extracts by reverse-phase chromatography also revealed more than one fraction with immunoreactivity. The immunoreactive material from the digestive tract was found to be less hydrophobic than insulin. Western blot analysis of the cerebral ganglia extract revealed more than one band with insulin immunoreactivity. Three of these bands had molecular masses very similar to those of vertebrate insulin, its subunits, and its precursor.