Expression profile and reproductive regulation of APGWamide in Pacific abalone (Haliotis discus hannai)

Neuropeptides in the central nervous system regulate reproductive activities in vertebrates. Ala-Pro-Gly-Trp-NH2 (APGWamide), a neuromediator expressed in the neural ganglia of mollusks, controls sexual maturation and reproduction. To clarify the role of APGWamide in sexual behavior regulation and gamete cell maturation in mollusks, we cloned the cDNA of APGWamide precursor (Hdh-APGWamide) and examined the spatiotemporal expression of the transcript in the Pacific abalone Haliotis discus hannai. The 222-amino acid sequence of the precursor deduced from the cDNA sequence showed typical features of gastropod APGWamide precursors. Phylogenetic analysis revealed that Hdh-APGWamide is classified with other gastropod APGWamide precursors, which form a separate branch from those of the bivalves. Hdh-APGWamide mRNA was highly expressed in the neural ganglia in both sexes. In females, the three ganglia (pleuro-pedal ganglion, PPG; branchial ganglion, and cerebral ganglion) showed similar expression in immature and mature animals, whereas in males, the level in the PPG only was higher at maturity (P < 0.05). In vivo injection of APGWamide or 5-hydroxytryptamine (10(-3)M) increased the frequency of spawning and the number of released sperm cells by mature males (P < 0.05), while concentrations above 10(-7)M enhanced germinal vesicle breakdown in fully developed cultured oocytes (P < 0.05). Thus, the phylogenetic branch of the APGWamide precursor gene in Haliotidae was separate from the other branches under the phylum Mollusca, and this gene exhibited ganglion-specific expression, indicating that it may induce final maturation and spawning in both sexes of Haliotis spp.