Identification and Expression Analysis of Tryptophan Hydroxylase in the Brain and Ventral Nerve Cord of Ragworm Neanthes japonica (Polychaeta, Annelida)

Abstract

Tryptophan hydroxylase (TPH) was stained in the central nervous system of the Neanthes japonica (Polychaeta, Annelida), using sheep antitryptophan hydroxylase antibody by the Streptavidin-Peroxidase immunohistochemical method and Colophony-Paraffin embedded section technique. The immunohistochemistry results revealed that the TPH is distributed in the brain and ventral nerve cord, which is consistent with that of serotonin (5-hydroxytryptamine, 5-HT) that labeled by antiserotonin antibody. Using the rapid amplification of cDNA ends (RACE) technique, TPH cDNA cloned from Neanthes japonica's central nervous system was 1778bp, which encodes predicted protein of 463 amino acid residues. The co-localization of TPH and 5-HT indicated that the specific TPH was responsible for the central serotonin synthesis in the central nervous system of annelida, TPH and 5-HT not only could be as the novel mutual corroboration marker to detect serotonergic neurons, but also provides the evidences for the evolution of aromatic amino acid hydroxylase genes. Anat Rec, 300:415-424,