4.4 Article

Molecular cloning of precursors for TEP-1 and TEP-2: The GGNG peptide-related peptides of a prosobranch gastropod, Thais clavigera

Journal

PEPTIDES
Volume 68, Issue -, Pages 72-82

Publisher

ELSEVIER SCIENCE INC
DOI: 10.1016/j.peptides.2014.10.009

Keywords

In situ hybridization; RT-PCR; Neuropeptide; Precursor; cDNA-library; Mollusk

Funding

  1. Japan Society for the Promotion of Science [16570063, 21248026]
  2. Sumitomo Foundation [053480]
  3. Grants-in-Aid for Scientific Research [21248026, 16570063] Funding Source: KAKEN

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TEP (Thais excitatory peptide)-1 and TEP-2 are molluscan counterparts of annelidan GGNG-peptides, identified in a neogastropod, Thais clavigera (Morishita et al., 2006). We have cloned two cDNAs encoding TEP-1 and TEP-2 precursor protein, respectively, by the standard molecular cloning techniques. Predicted TEP-1 precursor protein consists of 161 amino acids, while predicted TEP-2 precursor protein has 118 amino acids. Only a single copy of TEP was found on the respective precursor. The semi-quantitative RT-PCR showed that expression of TEP-1 was high in sub-esophageal, pleural, pedal and visceral ganglia, while it was low in supra-esophageal ganglion. By contrast, expression level of TEP-2 was high in pedal and visceral ganglia. In situ hybridization visualized different subsets of TEP-1 and TEP-2 expressing neurons in Thais ganglia. For example, supra-esophageal ganglion contained many TEP-2 expressing neuron, but not TEP-1 expressing ones. These results suggest that expression of TEP-1 and TEP-2 is differently regulated in the Thais ganglia. (C) 2014 Elsevier Inc. All rights reserved.

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