title: Molecular cloning of two distinct precursor genes of NdWFamide, a d-tryptophan-containing neuropeptide of the sea hare, Aplysia kurodai.
abstract: NdWFamide (NdWFa) is a D-tryptophan-containing cardioexcitatory neuropeptide in gastropod mollusks, such as Aplysia kurodai and Lymanea stagnalis. In this study, we have cloned two cDNA encoding distinct precursors for NdWFa from the abdominal ganglion of A. kurodai. One of the predicted precursor proteins consisted of 90 amino acids (NWF90), and the other consisted of 87 amino acids (NWF87). Both of the predicted precursor proteins have one NWFGKR sequence preceded by the N-terminal signal peptide. Sequential double staining by in situ hybridization (ISH) and immunostaining with anti-NdWFa antibody suggested that NdWFa-precursor and NdWFa peptide co-exist in neurons located in the right-upper quadrant region of the abdominal ganglion. In ISH, NWF90-specific signal and NWF87-specific one were found in different subsets of neurons in the abdominal ganglia of Aplysia. The expression level of NWF90 gene estimated by RT-PCR is much higher than that of NWF87 gene. These results suggest that NWF90 precursor is the major source of NdWFa in Aplysia ganglia.
authors.affiliation: Department of Biological Science, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-8526, Japan. fumi425@hiroshima-u.ac.jp
authors.forename: Fumihiro
authors.initials: F
authors.lastname: Morishita
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Furukawa
authors.forename: Osamu
authors.initials: O
authors.lastname: Matsushima
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia / Q000235:genetics*; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
chemical_list: D009842:Oligopeptides; C109335:asparaginyl-tryptophyl-phenylalaninamide