Identification of One Tachykinin- and Two Kinin-Related Peptides in the Brain of the White Shrimp,Penaeus vannamei

Abstract

This paper reports the purification of three myotropic neuropeptides from the white shrimp Penaeus vannamei. The central nervous systems of 3500 shrimps were extracted in an acidified solvent, after which four to five HPLC column systems were used to obtain pure peptides. A cockroach hindgut muscle contraction bioassay was used to monitor all collected fractions. The pure peptides were submitted to Edman degradation based automated microsequencing. Mass spectrometry and chemical synthesis confirmed the sequences. Ala-Pro-Ser-Gly-Phe-Leu-Gly-Met-Arg-NH2 (Pev-tachykinin, 934.1 Da) belongs to the tachykinin family with identified members in all vertebrate classes and some invertebrate classes: arthropods, annelids and molluscs. A very specific Pev-tachykinin antiserum was developed, which labels 4 neurosecretory cells in the brain. Ala-Ser-Phe-Ser-Pro-Trp-Gly-NH2 (Pev-kinin 1, 749.8 Da) and Asp-Phe-Ser-Ala-Trp-Ala-NH2 (Pev-kinin 2, 694.7 Da) are the first crustacean kinins. Pev-kinin 2 is the first kinin with a Trp-Ala-NH2 instead of a kinin-typical Trp-Gly-NH2 carboxyterminus.

Identification and characterization of a tachykinin-containing neuroendocrine organ in the commissural ganglion of the crabCancer productus

“…For example, 10 TRPs have been isolated from the cockroach Leucophea maderae Nässel, 1996, 1997), seven from the honeybee Apis mellifera (Takeuchi et al, 2003), five from the locust Locusta migratoria (Schoofs et al, 1990a(Schoofs et al, ,b, 1993, five from the fruit fly Drosophila melanogaster (Siviter et al, 2000) and three from the mosquito Culex salinarius (Meola et al, 1998). In contrast to the diversity of species-specific TRPs present in insects, only a single isoform is thought to be present in decapod crustaceans (Christie et al, 1997a;Nieto et al, 1998;Li et al, 2002a;Huybrechts et al, 2003;Yasuda-Kamatani and Yasuda, 2004). This peptide, APSGFLGMRamide or Cancer borealis tachykinin-related peptide Ia (CabTRP Ia), has been isolated from or is predicted by cDNA to be present in the crab Cancer borealis, the shrimp Panaeus vannamei, the chelate marine lobster Homarus americanus, the freshwater crayfish Procambarus clarkii and the spiny lobster Panulirus interruptus (Christie et al, 1997a;Nieto et al, 1998;Li et al, 2002a;Huybrechts et al, 2003;Yasuda-Kamatani and Yasuda, 2004).…”

Actions of kinin peptides in the stomatogastric ganglion of the crabCancer borealis

“…This antiserum binds with the carboxyl-terminal region of leucokinin I (Nässel et al, 1992). This region of the leucokinin I peptide is similar to the comparable region of the previously identified crustacean kinins, pevkinin (PevK) 1-6 (Nieto et al, 1998;Torfs et al, 1999). The tissue incubation in primary antiserum was followed by another five rinses over approximately 5·h in P-Triton.…”

“…The pevkinins were first isolated and identified in the shrimp Penaeus vannamei (Nieto et al, 1998;Torfs et al, 1999). A subset of the pevkinins (PevK-1, PevK-3, PevK-4) have the same carboxyl-terminal amino acid sequence as the insect kinins, including the leucokinins (Torfs et al, 1999).…”

“…Since then, more than 30 kinin family members have been isolated from many insect species (Holman et al, 1990;Schoofs et al, 1992;Hayes et al, 1994;Veenstra, 1994;Blackburn et al, 1995;Predel et al, 1997;Nieto et al, 1998;Terhzaz et al, 1999). The insect kinins all share the C-terminal amino acid sequence FX 1 X 2 WGamide, where X 1 is F, H, N, S or Y, and X 2 is A, P or S. The non-insect members share this sequence structure but for the terminal amino acid being changed from glycine to serine in lymnokinin (snail) and alanine in some pevkinin (shrimp) family members (Cox et al, 1997;Nieto et al, 1998).…”

“…Supporting the former possibility is the finding that kinin-like immunolabeling (KLI) occurs in the CNS of many arthopods, staining neuronal somata in the supraesophageal (brain), subesophageal, thoracic and abdominal ganglia (Torfs et al, 1999). In the shrimp Penaeus vannamei, pevkinin (PevK)-like immunolabeling includes a subset of neurosecretory cells in the brain and thoracic ganglion (Nieto et al, 1998). Supporting a hormonal action for kinin peptides is the finding that, in Leucophaea, leucokinin peptides influence the Malpighian tubes and the hindgut but neither structure is innervated by KLI axons (Nässel et al, 1992).…”