neuropeptide-f and corazonin-protein--insect

The cotton fleahopper, Pseudatomoscelis seriatus (Reuter), is an economically important pest of cotton, and increasing concerns over resistance, detrimental effects on beneficial insects and safety issues associated with traditional insecticide applications have led to an interest in research on novel, alternative strategies for control. One such approach requires a more basic understanding of the neurohormonal system that regulates important physiological properties of the fleahopper; e.g. the expression of specific messenger molecules such as neuropeptides. Therefore we performed a peptidomic study of neural tissues from the fleahopper which led to the first identification of the sequences of native peptide hormones. These peptide hormones include the following neuropeptides: corazonin, short neuropeptide F (sNPF), myosuppressin, CAPA-pyrokinin and CAPA-PVK peptides. The CAPA-pyrokinin, sNPF, and CAPA-PVK peptides represent novel sequences. A comparison of fleahopper neuropeptides with those of related heteropteran species indicates that they are quite different. The sNPF of P. seriatus shows, among others, a novel substitution of Leu with Phe within the C-terminal region; a modification that sets it apart from the known sNPFs of not only other Heteroptera but of other arthropod species as well. The identity of the neuropeptides native to the fleahopper can aid in the potential development of biostable, bioavailable mimetic agonists and antagonists capable of disrupting the physiological functions that these neuropeptides regulate.

Topics: Animals; Hemiptera; Insect Proteins; Neuropeptides; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The peptidome of the central nervous system of adult cabbage root fly, Delia radicum (L) was investigated using matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). Over twenty neuropeptides were identified from three different tissue sources, the combined brain/suboesophageal ganglion (SOG), the retrocerebral complex, and the thoracic-abdominal ganglion (TAG). A number of peptides were identified in all three tissues, including allatostatins, short neuropeptide F-like peptides, corazonin, a pyrokinin, and a myosuppressin. Adipokinetic hormone was restricted to the retrocerebral complex. Other peptides, including FMRFamides and sulfakinins were detected only in the brain/SOG and TAG. Some peptides, notably myoinhibitory peptides and tachykinins, which have been identified in other fly species, were not detected in any tissue sample. This study has structurally characterized for the first time, the neuropeptides from adult D. radicum.

Topics: Animals; Central Nervous System; Diptera; Insect Proteins; Neuropeptides; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tandem Mass Spectrometry

We performed the first comprehensive peptidomic analysis of neurohormones from hemipteran insects by analyzing the neuropeptides of two major neurohemal organs, namely the corpora cardiaca and abdominal perisympathetic organs. For the experiments we selected four related species of polyphagous stinkbugs (Pentatomidae), three of which are known to attack several important food crops. Peptide sequences were identified by MALDI-TOF mass spectrometry; tandem fragmentation of myosuppressin, sNPF, CAPA-periviscerokinins and pyrokinins revealed novel sequences not known from other insects so far. Most Leu/Ile and Glu/Lys ambiguities could be solved by either specific side-chain fragmentations or on-plate acetylation experiments. The identification of the specific sequences provides a solid basis for forthcoming pharmacological tests to study the neuroendocrine system of these pest insects. However, it should be mentioned in this context that the sequences of the peptides from different stinkbugs are likely not representative of Hemiptera in general. The forthcoming release of the genome from the reduviid Rhodnius prolixus will provide sufficient data to clear this point.

Topics: Amino Acid Sequence; Animals; Heteroptera; Insect Hormones; Insect Proteins; Molecular Sequence Data; Neuropeptides; Oligopeptides; Proteomics; Pyrrolidonecarboxylic Acid; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The Ixodoidea (ticks) are important vectors in the transmission of many human diseases; for example, the blacklegged tick Ixodes scapularis is the major vector in the transmission of Lyme disease, the most frequently reported vector-borne illness in the United States. The development of expressed sequence tags (ESTs) for ixodoidean cDNA libraries, and their public deposition, has generated a rich resource for protein discovery in members of this taxon, thereby providing an opportunity for better understanding the physiology and behavior of these disease vectors. Here, in silico searches of publicly accessible ESTs were conducted to identify transcripts encoding putative ixodoidean neuropeptide precursors, with the mature peptides contained within them predicted using online peptide processing programs and homology to known arthropod sequences. In total, 37 putative neuropeptide-encoding ESTs were identified from three ixodoidean species: I. scapularis (29 ESTs), Rhipicephalus microplus (seven ESTs) and Amblyomma americanum (one EST). Among those identified from I. scapularis were ones predicted to encode isoforms of corazonin, crustacean hyperglycemic hormone/ion transport peptide, diuretic hormone (both calcitonin- and corticotropin-releasing factor-like), FMRFamide-related peptide (both short neuropeptide F and sulfakinin subfamilies) orcokinin, proctolin, pyrokinin/periviscerokinin/pheromone biosynthesis activating neuropeptide, SIFamide, and tachykinin-related peptide. Collectively, 80 distinct ixodoidean neuropeptides were characterized from the identified precursors. These results not only expand greatly the number of known/predicted ixodoidean neuropeptides, but also provide a strong foundation for future molecular and physiological investigations of peptidergic control in this important group of disease-transmitting arthropods.

Topics: Amino Acid Sequence; Animals; Base Sequence; Databases, Genetic; Databases, Nucleic Acid; Expressed Sequence Tags; Gene Library; Insect Proteins; Molecular Sequence Data; Neuropeptides; Oligopeptides; Sequence Alignment; Ticks

Activation of G protein-coupled receptors (GPCR) leads to the recruitment of beta-arrestins. By tagging the beta-arrestin molecule with a green fluorescent protein, we can visualize the activation of GPCRs in living cells. We have used this approach to de-orphan and study 11 GPCRs for neuropeptide receptors in Drosophila melanogaster. Here we verify the identities of ligands for several recently de-orphaned receptors, including the receptors for the Drosophila neuropeptides proctolin (CG6986), neuropeptide F (CG1147), corazonin (CG10698), dFMRF-amide (CG2114), and allatostatin C (CG7285 and CG13702). We also de-orphan CG6515 and CG7887 by showing these two suspected tachykinin receptor family members respond specifically to a Drosophila tachykinin neuropeptide. Additionally, the translocation assay was used to de-orphan three Drosophila receptors. We show that CG14484, encoding a receptor related to vertebrate bombesin receptors, responds specifically to allatostatin B. Furthermore, the pair of paralogous receptors CG8985 and CG13803 responds specifically to the FMRF-amide-related peptide dromyosuppressin. To corroborate the findings on orphan receptors obtained by the translocation assay, we show that dromyosuppressin also stimulated GTPgammaS binding and inhibited cAMP by CG8985 and CG13803. Together these observations demonstrate the beta-arrestin-green fluorescent protein translocation assay is an important tool in the repertoire of strategies for ligand identification of novel G protein-coupled receptors.

Topics: Animals; Arrestins; beta-Arrestins; Cell Line; Cloning, Molecular; Cyclic AMP; Dose-Response Relationship, Drug; Drosophila; Drosophila Proteins; FMRFamide; Green Fluorescent Proteins; Guanosine 5'-O-(3-Thiotriphosphate); Humans; Insect Hormones; Insect Proteins; Ligands; Luminescent Proteins; Microscopy, Confocal; Neuropeptides; Oligopeptides; Peptides; Protein Transport; Receptors, G-Protein-Coupled; Receptors, Neuropeptide; Receptors, Peptide; Receptors, Tachykinin; Transfection