allatostatin-1 and crustacean-cardioactive-peptide

The aquaculture of crabs from the genus Scylla is of increasing economic importance for many Southeast Asian countries. Expansion of Scylla farming has led to increased efforts to understand the physiology and behavior of these crabs, and as such, there are growing molecular resources for them. Here, publicly accessible Scylla olivacea transcriptomic data were mined for putative peptide-encoding transcripts; the proteins deduced from the identified sequences were then used to predict the structures of mature peptide hormones. Forty-nine pre/preprohormone-encoding transcripts were identified, allowing for the prediction of 187 distinct mature peptides. The identified peptides included isoforms of adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin B, allatostatin C, bursicon β, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone/molt-inhibiting hormone, diuretic hormone 31, eclosion hormone, FMRFamide-like peptide, HIGSLYRamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, pyrokinin, red pigment concentrating hormone, RYamide, short neuropeptide F, SIFamide and tachykinin-related peptide, all well-known neuropeptide families. Surprisingly, the tissue used to generate the transcriptome mined here is reported to be testis. Whether or not the testis samples had neural contamination is unknown. However, if the peptides are truly produced by this reproductive organ, it could have far reaching consequences for the study of crustacean endocrinology, particularly in the area of reproductive control. Regardless, this peptidome is the largest thus far predicted for any brachyuran (true crab) species, and will serve as a foundation for future studies of peptidergic control in members of the commercially important genus Scylla.

Topics: Amino Acid Sequence; Animals; Arthropod Proteins; Brachyura; FMRFamide; Invertebrate Hormones; Male; Nerve Tissue Proteins; Neuropeptides; Peptide Hormones; Proteome; Testis; Transcriptome

Our previous report showed that the pars intercerebralis (PI)-ablated cockroach, Periplaneta americana (PIX), exhibited hypertrophy and a significant increase in α-amylase and protease activities in the midgut under constant darkness (DD). Bath-applied crustacean cardioactive peptide (CCAP) and allatostatin (AST) stimulated α-amylase and protease activities in the dissected midgut cultured in medium. However, the functional relationship and regulatory mechanism between the brain, particularly the pars intercerebralis and the midgut digestive activity remain to be investigated. Here, we investigated the immunohistochemical reactivities (IHCr) against CCAP and AST in the midgut of cockroach subjected to the above operation (PIX-DD). Three types of IHCr cells were observed in both the muscle layer and the epithelium: (1) CCAP-ir only, (2) AST-ir only and (3) both reactivities are colocalized. The number of all three types increased intensively after PIX under DD compared with that of sham operated control that was kept under constant condition (CNT-DD), indicating that the PI suppresses the expression of CCAP and AST in the midgut epithelium. We also showed that co-administration of CCAP and AST to the midgut caused increases of 1.5-fold and 1.4-fold for α-amylase and protease activities, respectively, compared with application of either peptide above. On the other hand, CCAP-ir in the muscle layer was more strongly expressed but AST-ir was suppressed in PIX-DD. While these peptides showed opposite effects on spontaneous contraction, when epithelially released, these peptides both activated the digestive enzyme system. Overall, up-regulated AST-6 and down-regulated CCAP in the stomatogastric nerve in the muscle layer produce the same end result, that is, stimulation of digestive activity (hypertrophy) via both enzyme activation and the retarded peristalsis that leads to increased throughput time.

Topics: alpha-Amylases; Animals; Cerebrum; Digestive System; Gene Expression Regulation; Insect Proteins; Neuropeptides; Peptide Hydrolases; Periplaneta; Up-Regulation

The genome of the spider mite was prospected for the presence of genes coding neuropeptides, neurohormones and their putative G-protein coupled receptors. Fifty one candidate genes were found to encode neuropeptides or neurohormones. These include all known insect neuropeptides and neurohormones, with the exception of sulfakinin, corazonin, neuroparsin and PTTH. True orthologs of adipokinetic hormone (AKH) were neither found, but there are three genes encoding peptides similar in structure to both AKH and the AKH-corazonin-related peptide. We were also unable to identify the precursors for pigment dispersing factor (PDF) or the recently discovered trissin. However, the spider mite probably does have such genes, as we found their putative receptors. A novel arthropod neuropeptide gene was identified that shows similarity to previously described molluscan neuropeptide genes and was called EFLamide. A total of 65 putative neuropeptide GPCR genes were also identified, of these 58 belong to the A-family and 7 to the B-family. Phylogenetic analysis showed that 50 of them are closely related to insect GPCRs, which allowed the identification of their putative ligand in 39 cases with varying degrees of certainty. Other spider mite GPCRs however have no identifiable orthologs in the genomes of the four holometabolous insect species best analyzed. Whereas some of the latter have orthologs in hemimetabolous insect species, crustaceans or ticks, for others such arthropod homologs are currently unknown.

Topics: Amino Acid Sequence; Animals; Arthropod Proteins; Insect Hormones; Insulins; Invertebrate Hormones; Molecular Sequence Data; Nerve Tissue Proteins; Neuropeptides; Neurotransmitter Agents; Oligopeptides; Receptors, G-Protein-Coupled; Tetranychidae

While numerous investigations have focused on the identification of neuropeptides in arthropods, most have been conducted on members of the Hexapoda or Crustacea, and little is currently known about those in the Chelicerata. Here, publicly accessible expressed sequence tags (ESTs) were mined for putative chelicerate neuropeptide-encoding transcripts; the peptides encoded by the ESTs were deduced using on-line peptide prediction programs and homology to known isoforms. Fifty-eight ESTs representing eight peptide families/subfamilies were identified using this strategy. Of note was the prediction of the first authentic chelicerate C-type allatostatin, pQIRYHQCYFNPISCF, from the mite Tetranychus urticae, as well as the prediction a novel allatostatin CC peptide, GEGKMFWRCYFNAVSCF, from both the tick Amblyomma variegatum and the scorpion Mesobuthus gibbosus. Also identified from T. urticae were authentic crustacean cardioactive peptide (CCAP), several peptides belonging to the crustacean hyperglycemic hormone/ion transport peptide superfamily, members of the calcitonin-like diuretic hormone/diuretic hormone 31 family, and several FMRFamide-like peptides, specifically members of the neuropeptide F (NPF) and short neuropeptide F subfamilies. To the best of our knowledge the identifications of CCAP and NPF in T. urticae are the first for the Chelicerata. In addition, several novel orcokinins were identified from the scorpion Scorpiops jendeki and the spider Loxosceles laeta; in S. jendeki previously unknown isoforms of SIFamide, ESRNPPLNGSMFamide and ESKNPPLNGSMFamide, were also predicted. Taken collectively, the data presented in our study expand the catalog of known chelicerate neuropeptides and provide a foundation for future physiological studies of them in these animals.

Topics: Animals; Arthropods; Computational Biology; Expressed Sequence Tags; Neuropeptides

The cladoceran crustacean Daphnia pulex has emerged as a model species for many biological fields, in particular environmental toxicology and toxicogenomics. Recently, this species has been the subject of an extensive transcriptome project, resulting in the generation and public deposition of over 150,000 expressed sequence tags (ESTs). This resource makes D. pulex an excellent model for protein discovery using bioinformatics. Here, in silico searches of the D. pulex EST database were conducted to identify transcripts encoding putative peptide precursors. Moreover, the mature peptides contained within the deduced prepro-hormones were predicted using online peptide processing programs and homology to known arthropod isoforms. In total, 63 putative peptide-encoding ESTs were identified encompassing 14 distinct peptide families/subfamilies: A-type allatostatin, B-type allatostatin, C-type allatostatin, bursicon (both alpha and beta subunit peptides), crustacean cardioactive peptide (CCAP), crustacean hyperglycemic hormone (CHH)/ion transport peptide (both CHH- and moult-inhibiting hormone-like subfamilies), diuretic hormone (calcitonin-like), ecdysis-triggering hormone (ETH), FMRFamide (both neuropeptide F and short neuropeptide F subfamilies), orcokinin and pigment dispersing hormone. From these transcripts, the structures of 76 full-length/partial peptides were predicted, which included the first C-type allatostatin-like peptide identified from a crustacean, the first crustacean calcitonin-like diuretic hormone, an undescribed CCAP isoform, two hitherto unknown ETH variants, and two new orcokinins. Neuronal localization of several of the identified peptide families was confirmed using immunohistochemitry (i.e. A-type allatostatin, CCAP, FMRFamide and PDH). In addition, immunohistochemical analyses identified other putative neuropeptides for which no ESTs had been found (i.e. corazonin, insect kinin, proctolin, red pigment concentrating hormone, SIFamide, sulfakinin and tachykinin-related peptide). Collectively, the data presented here not only catalog an extensive array of putative D. pulex peptide paracrines/hormones, but also provide a strong foundation for future investigations of the effects of environmental/anthropogenic stressors on peptidergic control in this model organism.

Topics: Amino Acid Sequence; Animals; Arthropod Proteins; Central Nervous System; Computational Biology; Daphnia; Expressed Sequence Tags; FMRFamide; Gene Expression Profiling; Immunohistochemistry; Intercellular Signaling Peptides and Proteins; Invertebrate Hormones; Molecular Sequence Data; Nerve Tissue Proteins; Neuropeptides; Paracrine Communication; Sequence Alignment

The pericardial organs (POs) are a pair of neurosecretory organs that surround the crustacean heart and release neuromodulators into the hemolymph. In adult crustaceans, the POs are known to contain a wide array of peptide and amine modulators. However, little is known about the modulatory content of POs early in development. We characterize the morphology and modulatory content of pericardial organs in the embryonic lobster, Homarus americanus. The POs are well developed by midway through embryonic (E50) life and contain a wide array of neuromodulatory substances. Immunoreactivities to orcokinin, extended FLRFamide peptides, tyrosine hydroxylase, proctolin, allatostatin, serotonin, Cancer borealis tachykinin-related peptide, cholecystokinin, and crustacean cardioactive peptide are present in the POs by approximately midway through embryonic life. There are two classes of projection patterns to the POs. Immunoreactivities to orcokinin, extended FLRFamide peptides, and tyrosine hydroxylase project solely from the subesophageal ganglion (SEG), whereas the remaining modulators project from the SEG as well as from the thoracic ganglia. Double-labeling experiments with a subset of modulators did not reveal any colocalized peptides in the POs. These results suggest that the POs could be a major source of neuromodulators early in development.

Topics: Animals; Heart; Nephropidae; Nervous System; Neural Pathways; Neuropeptides; Neurosecretory Systems; Neurotransmitter Agents; Oligopeptides; Serotonin; Tyrosine 3-Monooxygenase