ABSTRACT
OBJECTIVE: It has been previously shown that brain-derived neurotrophic factor is linked with various types of cancer. Brain-derived neurotrophic factor is found to be highly expressed in multiple human cancers and associated with tumor growth, invasion, and metastasis. Adipokinetic hormones are functionally related to the vertebrate glucagon, as they have similar functionalities that manage the nutrient-dependent secretion of these two hormones. Migrasomes are new organelles that contain numerous small vesicles, which aid in transmitting signals between the migrating cells. Therefore, the aim of this study was to investigate the effects of Anax imperator adipokinetic hormone on brain-derived neurotrophic factor expression and ultrastructure of cells in the C6 glioma cell line. METHODS: The rat C6 glioma cells were treated with concentrations of 5 and 10 Anax imperator adipokinetic hormone for 24 h. The effects of the Anax imperator adipokinetic hormone on the migrasome formation and brain-derived neurotrophic factor expression were analyzed using immunocytochemistry and transmission electron microscope. RESULTS: The rat C6 glioma cells of the 5 and 10 µM Anax imperator adipokinetic hormone groups showed significantly high expressions of brain-derived neurotrophic factor and migrasomes numbers, compared with the control group. CONCLUSION: A positive correlation was found between the brain-derived neurotrophic factor expression level and the formation of migrasome, which indicates that the increased expression of brain-derived neurotrophic factor and the number of migrasomes may be involved to metastasis of the rat C6 glioma cell line induced by the Anax imperator adipokinetic hormone. Therefore, the expression of brain-derived neurotrophic factor and migrasome formation may be promising targets for preventing tumor proliferation, invasion, and metastasis in glioma.
Subject(s)
Brain-Derived Neurotrophic Factor , Glioma , Oligopeptides , Pyrrolidonecarboxylic Acid , Glioma/metabolism , Glioma/pathology , Animals , Brain-Derived Neurotrophic Factor/metabolism , Rats , Cell Line, Tumor , Pyrrolidonecarboxylic Acid/analogs & derivatives , Pyrrolidonecarboxylic Acid/metabolism , Oligopeptides/pharmacology , Insect Hormones/metabolism , Cell Movement/drug effects , Immunohistochemistry , Brain Neoplasms/metabolism , Brain Neoplasms/pathology , Organelles/metabolism , Organelles/drug effects , Organelles/ultrastructureABSTRACT
Allatotropin is a pleiotropic peptide originally characterized in insects. The existence of AT neuropeptide signaling was proposed in other invertebrates. In fact, we previously proposed the presence of an AT-like system regulating feeding behavior in Hydra sp. Even in insects, the information about the AT signaling pathway is incomplete. The aim of this study is to analyze the signaling cascade activated by AT in Hydra plagiodesmica using a pharmacological approach. The results show the involvement of Ca2+ and IP3 signaling in the transduction pathway of the peptide. Furthermore, we confirm the existence of a GPCR system involved in this pathway, that would be coupled to a Gq subfamily of Gα protein, which activates a PLC, inducing an increase in IP3 and cytosolic Ca2+. To the best of our knowledge, this work represents the first in vivo approach to study the overall signaling pathway and intracellular events involved in the myoregulatory effect of AT in Hydra sp.
Subject(s)
Calcium Signaling , Hydra/metabolism , Insect Hormones/metabolism , Neuropeptides/metabolism , Orexins/metabolism , Receptors, G-Protein-Coupled/metabolism , Animals , Calcium/metabolism , Calcium Signaling/drug effects , Enzyme Inhibitors/pharmacology , Estrenes/pharmacology , GTP-Binding Proteins/metabolism , Indoles/pharmacology , Inositol 1,4,5-Trisphosphate Receptors/metabolism , Maleimides/pharmacology , Melitten/pharmacology , Protein Kinase C/antagonists & inhibitors , Protein Kinase C/metabolism , Pyrrolidinones/pharmacology , Type C Phospholipases/antagonists & inhibitors , Type C Phospholipases/metabolismABSTRACT
Helicoverpa armigera (Hübner) regularly requires quick deployment of control practices due to its potential of causing yield loss of processing tomatoes. The monitoring of H. armigera adults using stick traps lured with sex pheromone was correlated to egg densities, larvae, and percentage of damaged fruits, aiming to subsidize control decisions only using the number of males caught in the traps since surveying pest stages or damaged fruits is laborious and costly. Surveys were carried out on 11 tomato fields (ca. 80 to 120 hectares each) for processing during the 2015 and 2016 crop seasons. The survey was initiated at the flowering stage with traps set up along the field borders 100 m distant each. Evaluations of adult male capture in the traps were conducted twice a week for 14 and 10 weeks during the 2015 and 2016 seasons, respectively. Simultaneously, 70 tomato plants were fully inspected for eggs, larvae, and damaged fruits per area using a rate of 10 plants per point of trap setting in each area. Overall capture of male moths correlated to subsequent eggs, larvae, and damaged fruits. Based on the outcome of the regression analysis between male moths captured and percentage of damaged fruits, the control decision is suggested when three to six male moths are captured per trap during one week. The monitoring of adults using pheromone traps help deploy chemical or biological control methods to restrain H. armigera population reaching economic threshold levels.(AU)
Subject(s)
Lepidoptera/pathogenicity , Solanum lycopersicum , Insect Hormones/biosynthesisABSTRACT
Helicoverpa armigera (Hübner) regularly requires quick deployment of control practices due to its potential of causing yield loss of processing tomatoes. The monitoring of H. armigera adults using stick traps lured with sex pheromone was correlated to egg densities, larvae, and percentage of damaged fruits, aiming to subsidize control decisions only using the number of males caught in the traps since surveying pest stages or damaged fruits is laborious and costly. Surveys were carried out on 11 tomato fields (ca. 80 to 120 hectares each) for processing during the 2015 and 2016 crop seasons. The survey was initiated at the flowering stage with traps set up along the field borders 100 m distant each. Evaluations of adult male capture in the traps were conducted twice a week for 14 and 10 weeks during the 2015 and 2016 seasons, respectively. Simultaneously, 70 tomato plants were fully inspected for eggs, larvae, and damaged fruits per area using a rate of 10 plants per point of trap setting in each area. Overall capture of male moths correlated to subsequent eggs, larvae, and damaged fruits. Based on the outcome of the regression analysis between male moths captured and percentage of damaged fruits, the control decision is suggested when three to six male moths are captured per trap during one week. The monitoring of adults using pheromone traps help deploy chemical or biological control methods to restrain H. armigera population reaching economic threshold levels.
Subject(s)
Insect Hormones/biosynthesis , Lepidoptera/pathogenicity , Solanum lycopersicumABSTRACT
We describe a pair of labial gland lobes on either side of the retrocerebral complex in the head of the Australian black field cricket, Teleogryllus commodus Walker. As the retrocerebral complex includes the corpora cardiaca and corpora allata, hormones secreted by these glands can be absorbed by these lobes. These lobes of the labial gland are connected to the thoracic lobes via a relatively long duct that enters the main duct draining the thoracic lobes. Measurement of the flow rate of dye from head to thorax in the ducts is rapid, suggesting that these glands may serve as a transport system into the thoracic region. Both serotonin and adipokinetic hormone are shown to be present in the lobes near the retrocerebral complex and the ducts of the thoracic lobes, but whether this connection between the head and thorax acts as a hormone transporter is still unclear.
Subject(s)
Gryllidae/anatomy & histology , Animals , Endocrine Glands/anatomy & histology , Gryllidae/metabolism , Head/anatomy & histology , Insect Hormones/metabolismABSTRACT
BACKGROUND: In insects, continuous growth requires the periodic replacement of the exoskeleton. Once the remains of the exoskeleton from the previous stage have been shed during ecdysis, the new one is rapidly sclerotized (hardened) and melanized (pigmented), a process collectively known as tanning. The rapid tanning that occurs after ecdysis is critical for insect survival, as it reduces desiccation, and gives the exoskeleton the rigidity needed to support the internal organs and to provide a solid anchor for the muscles. This rapid postecdysial tanning is triggered by the "tanning hormone", bursicon. Since bursicon is released into the hemolymph, it has naturally been assumed that it would act on the epidermal cells to cause the tanning of the overlying exoskeleton. RESULTS: Here we investigated the site of bursicon action in Drosophila by examining the consequences on tanning of disabling the bursicon receptor (encoded by the rickets gene) in different tissues. To our surprise, we found that rapid tanning does not require rickets function in the epidermis but requires it instead in peptidergic neurons of the ventral nervous system (VNS). Although we were unable to identify the signal that is transmitted from the VNS to the epidermis, we show that neurons that express the Drosophila insulin-like peptide ILP7, but not the ILP7 peptide itself, are involved. In addition, we found that some of the bursicon targets involved in melanization are different from those that cause sclerotization. CONCLUSIONS: Our findings show that bursicon does not act directly on the epidermis to cause the tanning of the overlying exoskeleton but instead requires an intermediary messenger produced by peptidergic neurons within the central nervous system. Thus, this work has uncovered an unexpected layer of control in a process that is critical for insect survival, which will significantly alter the direction of future research aimed at understanding how rapid postecdysial tanning occurs.
Subject(s)
Animal Shells/physiology , Drosophila/physiology , Insect Hormones/metabolism , Invertebrate Hormones/metabolism , Animals , Drosophila Proteins/metabolism , Epidermis/physiology , Female , Male , Neuropeptides/metabolismABSTRACT
Peptidic messengers constitute a highly diversified group of intercellular messengers widely distributed in nature that regulate a great number of physiological processes in Metazoa. Being crucial for life, it seem that they have appeared in the ancestral group from which Metazoa evolved, and were highly conserved along the evolutionary process. Peptides act mainly through G-protein coupled receptors (GPCRs), a family of transmembrane molecules. GPCRs are also widely distributed in nature being present in metazoan, but also in Choanoflagellata and Fungi. Among GPCRs, the Allatotropin/Orexin (AT/Ox) family is particularly characterized by the presence of the DRW motif in the second intracellular loop (IC Loop 2), and seems to be present in Cnidaria, Placozoa and in Bilateria, suggesting that it was present in the common ancestor of Metazoa. Looking for the evolutionary history of this GPCRs we searched for corresponding sequences in public databases. Our results suggest that AT/Ox receptors were highly conserved along evolutionary process, and that they are characterized by the presence of the E/DRWYAI motif at the IC Loop 2. Phylogenetic analyses show that AT/Ox family of receptors reflects evolutionary relationships that agree with current phylogenetic understanding in Actinopterygii and Sauropsida, including also the largely discussed position of Testudines.
Subject(s)
Insect Hormones/genetics , Neuropeptides/genetics , Orexins/genetics , Animals , Biological Evolution , Classification/methods , Cnidaria/classification , Cnidaria/genetics , Databases, Genetic , Evolution, Molecular , Insect Hormones/metabolism , Neuropeptides/metabolism , Orexins/metabolism , Phylogeny , Placozoa/classification , Placozoa/genetics , Receptors, G-Protein-Coupled/genetics , Sequence Analysis, DNA , Vertebrates/geneticsABSTRACT
Ecdysis is a vital process for insects, during which they shed the old cuticle in order to emerge as the following developmental stage. Given its relevance for survival and reproduction, ecdysis is tightly regulated by peptidic hormones that conform an interrelated neuromodulatory network. This network was studied in species that undergo a complete metamorphosis, but not in hemimetabola. In a recent work, we demonstrated that orcokinin neuropeptides are essential for ecdysis to occur in the kissing bug Rhodnius prolixus. Here we performed gene silencing, quantitative PCR and in vitro treatments in order to study the interrelationships between RhoprOKs and hormones such as ecdysis triggering hormone, corazonin, eclosion hormone, crustacean cardioactive peptide and ecdysone. Our results suggest that RhoprOKs directly or indirectly regulate the expression of other genes. Whereas RhoprOKA is centrally involved in the regulation of gene expression, RhoprOKB is implicated in processes related to midgut physiology. Therefore, we propose that the different transcripts encoded in RhoprOK gene could integrate signaling cues, in order to coordinate the nutritional state with development and ecdysis. Given the emerging data that point to OKs as important factors for survival and reproduction, they could be candidates in the search for new insect management strategies based on neuroendocrine targets.
Subject(s)
Neuropeptides/physiology , Rhodnius/physiology , Animals , Gene Silencing , Insect Hormones/genetics , Insect Hormones/metabolism , Molting/genetics , Molting/physiology , Protein Precursors/genetics , Protein Precursors/metabolism , Rhodnius/geneticsABSTRACT
Haematophagous insects can ingest large quantities of blood in a single meal producing a large quantity of urine in the following hours to eliminate the excess of water and mineral ions incorporated. The excretory activity of the Malpighian tubules is facilitated by an increase in haemolymph circulation as a result of the intensification of aorta contractions, combined with an increase of anterior midgut peristaltic waves. We have recently shown that haemolymph circulation during post-prandial diuresis is modulated by the synergistic activity of allatotropin (AT) and serotonin, resulting in an increase in aorta and crop contraction rates. In the present study we describe the antagonistic effect of allatostatin-C (AST-C) on the increase of aorta frequency of contractions induced by serotonin/AT in Rhodnius prolixus. The administration of AST-C counteracted the increase in the frequency induced by the treatment with serotonin/AT, but did not affect the increase in frequency induced by the administration of serotonin alone, suggesting that AST-C is altering the synergism between serotonin and AT. Furthermore, the administration of AST-C during post-prandial diuresis decreases the number of peristaltic waves of the anterior midgut. The AST-C putative receptor is expressed in the hindgut, midgut and dorsal vessel, three critical organs involved in post-prandial diuresis. All together these findings provide evidence that AST-C plays a key role as a myoregulatory and cardioregulatory peptide in R. prolixus.
Subject(s)
Insect Hormones/antagonists & inhibitors , Muscle Contraction/drug effects , Neuropeptides/antagonists & inhibitors , Neuropeptides/pharmacology , Rhodnius , Serotonin Antagonists/pharmacology , Serotonin/pharmacology , Animals , Aorta/drug effects , Aorta/physiology , Diuresis/drug effects , Drug Interactions , Female , Insect Hormones/pharmacology , Malpighian Tubules/drug effects , Malpighian Tubules/metabolism , Myocardial Contraction/drug effects , Postprandial Period/drug effects , Rhodnius/drug effects , Rhodnius/physiologyABSTRACT
Adipokinetic hormone (AKH) has been associated with the control of energy metabolism in a large number of arthropod species due to its role on the stimulation of lipid, carbohydrate and amino acid mobilization/release. In the insect Rhodnius prolixus, a vector of Chagas' disease, triacylglycerol (TAG) stores must be mobilized to sustain the metabolic requirements during moments of exercise or starvation. Besides the recent identification of the R. prolixus AKH peptide, other components required for the AKH signaling cascade and its mode of action remain uncharacterized in this insect. In the present study, we identified and investigated the expression profile of the gene encoding the AKH receptor of R. prolixus (RhoprAkhr). This gene is highly conserved in comparison to other sequences already described and its transcript is abundant in the fat body and the flight muscle of the kissing bug. Moreover, RhoprAkhr expression is induced in the fat body at moments of increased TAG mobilization; the knockdown of this gene resulted in TAG accumulation both in fat body and flight muscle after starvation. The inhibition of Rhopr-AKHR transcription as well as the treatment of insects with the peptide Rhopr-AKH in its synthetic form altered the transcript levels of two genes involved in lipid metabolism, the acyl-CoA-binding protein-1 (RhoprAcbp1) and the mitochondrial glycerol-3-phosphate acyltransferase-1 (RhoprGpat1). These results indicate that the AKH receptor is regulated at transcriptional level and is required for TAG mobilization under starvation. In addition to the classical view of AKH as a direct regulator of enzymatic activity, we propose here that AKH signaling may account for the regulation of nutrient metabolism by affecting the expression profile of target genes.
Subject(s)
Insect Hormones/metabolism , Oligopeptides/metabolism , Pyrrolidonecarboxylic Acid/analogs & derivatives , Receptors, Cell Surface/genetics , Rhodnius/genetics , Rhodnius/metabolism , Triglycerides/metabolism , Animals , Fat Body/metabolism , Flight, Animal , Food Deprivation , Gene Expression Regulation , Muscle, Skeletal/metabolism , Pyrrolidonecarboxylic Acid/metabolism , Receptors, Cell Surface/metabolism , Signal TransductionABSTRACT
Insect growth is punctuated by molts, during which the animal produces a new exoskeleton. The molt culminates in ecdysis, an ordered sequence of behaviors that causes the old cuticle to be shed. This sequence is activated by Ecdysis triggering hormone (ETH), which acts on the CNS to activate neurons that produce neuropeptides implicated in ecdysis, including Eclosion hormone (EH), Crustacean cardioactive peptide (CCAP) and Bursicon. Despite more than 40â years of research on ecdysis, our understanding of the precise roles of these neurohormones remains rudimentary. Of particular interest is EH; although it is known to upregulate ETH release, other roles for EH have remained elusive. We isolated an Eh null mutant in Drosophila and used it to investigate the role of EH in larval ecdysis. We found that null mutant animals invariably died at around the time of ecdysis, revealing an essential role in its control. Further analyses showed that these animals failed to express the preparatory behavior of pre-ecdysis while directly expressing the motor program of ecdysis. Although ETH release could not be detected, the lack of pre-ecdysis could not be rescued by injections of ETH, suggesting that EH is required within the CNS for ETH to trigger the normal ecdysial sequence. Using a genetically encoded calcium probe, we showed that EH configured the response of the CNS to ETH. These findings show that EH plays an essential role in the Drosophila CNS in the control of ecdysis, in addition to its known role in the periphery of triggering ETH release.
Subject(s)
Drosophila melanogaster/growth & development , Drosophila melanogaster/genetics , Insect Hormones/genetics , Molting , Alleles , Animals , Behavior, Animal , Hemizygote , Injections , Insect Hormones/metabolism , Larva/growth & development , Mutation/genetics , Neurons/metabolism , Neuropeptides/metabolismABSTRACT
Haematophagous insects can ingest large quantities of blood in a single meal and eliminate high volumes of urine in the next few hours. This rise in diuresis is possible because the excretory activity of the Malpighian tubules is facilitated by an increase in haemolymph circulation as a result of intensification of aorta contractions combined with an increase of the anterior midgut peristaltic waves. It has been previously described that haemolymph circulation during post-prandial diuresis is stimulated by the synergistic activity of allatotropin (AT) and serotonin in the kissing bug Triatoma infestans; resulting in an increase in aorta contractions. In the same species, AT stimulates anterior midgut and rectum muscle contractions to mix urine and feces and facilitate the voiding of the rectum. Furthermore, levels of AT in midgut and Malpighian tubules increased in the afternoon when insects are getting ready for nocturnal feeding. In the present study we describe the synergistic effect of AT and serotonin increasing the frequency of contractions of the aorta in Rhodnius prolixus. The basal frequency of contractions of the aorta in the afternoon is higher that the observed during the morning, suggesting the existence of a daily rhythmic activity. The AT receptor is expressed in the rectum, midgut and dorsal vessel, three critical organs involved in post-prandial diuresis. All together these findings provide evidence that AT plays a role as a myoregulatory and cardioacceleratory peptide in R. prolixus.
Subject(s)
Insect Hormones/physiology , Neuropeptides/physiology , Rhodnius/physiology , Animals , Aorta/physiology , Circadian Rhythm , Insect Proteins/metabolism , Male , Muscle Contraction , Muscle, Smooth/physiology , Organ Specificity , Receptors, Peptide/metabolism , Serotonin/physiology , VasoconstrictionABSTRACT
In most species of social insect the queen signals her presence to her workers via pheromones. Worker responses to queen pheromones include retinue formation around the queen, inhibition of queen cell production and suppression of worker ovary activation. Here we show that the queen signal of the Brazilian stingless bee Friesella schrottkyi is a mixture of cuticular hydrocarbons. Stingless bees are therefore similar to ants, wasps and bumble bees, but differ from honey bees in which the queen's signal mostly comprises volatile compounds originating from the mandibular glands. This shows that cuticular hydrocarbons have independently evolved as the queen's signal across multiple taxa, and that the honey bees are exceptional. We also report the distribution of four active queen-signal compounds by Matrix-assisted laser desorption/ionization (MALDI) imaging. The results indicate a relationship between the behavior of workers towards the queen and the likely site of secretion of the queen's pheromones.
Subject(s)
Bees/physiology , Insect Hormones/physiology , Ovary/physiology , Animal Communication , Animals , Female , Social BehaviorABSTRACT
Bombyx mori BmHRP28 and BmPSI, which belong to the family of RNA-binding proteins, have been identified binding to the female-specific exon 4 of the sex-determining gene Bmdsx pre-mRNA. However, the relationships between BmHRP28 and BmPSI still remain unclear. In this study, we carried out yeast two-hybrid (Y2H) and co-immunoprecipitation (Co-IP) analyses to address them. Y2H analysis showed that there was little or no direct binding between the BmHRP28 and BmPSI proteins. Also, the Co-IP experiments revealed that BmHRP28 and BmPSI coexisted in a multiprotein complex. Our results suggested that BmHRP28 and BmPSI form a muliprotein complex to regulate the splicing of Bmdsx pre-mRNA, but are not directly bound to each other. In an effort to find other regulatory factors in the multiprotein complex, we constructed a silkworm Y2H cDNA library of male early embryo. By Y2H screening, we identified an RNA-binding protein BmSPX, a putative component of the spliceosome, binding to BmPSI. These results indicated that BmHRP28 and BmPSI make up a spliceosome complex to regulate Bmdsx splicing and that BmSPX is another potential protein involved in this process. Our study provides some clues to better understand the mechanism of sex determination in the silkworm.
Subject(s)
Bombyx/genetics , Heterogeneous-Nuclear Ribonucleoproteins/genetics , Insect Hormones/genetics , Insect Proteins/genetics , RNA-Binding Proteins/genetics , Sex Determination Processes , Alternative Splicing , Amino Acid Sequence , Animals , Bombyx/growth & development , Embryo, Nonmammalian , Epistasis, Genetic , Exons , Female , Gene Library , Heterogeneous-Nuclear Ribonucleoproteins/metabolism , Insect Hormones/metabolism , Insect Proteins/metabolism , Male , Molecular Sequence Data , Protein Binding , RNA Precursors/genetics , RNA Precursors/metabolism , RNA-Binding Proteins/metabolism , Sequence Alignment , Two-Hybrid System TechniquesABSTRACT
Objetivo. Examinar la investigación hecha en México sobre los determinantes sociales de la salud (DSS) durante el periodo 2005-2012 con base en la caracterización del sistema nacional de investigación en salud y la producción científica sobre este tema. Material y métodos. Análisis en dos etapas: revisión documental de fuentes oficiales sobre investigación en salud en México y búsqueda sistemática de literatura sobre DSS. Resultados. Los DSS fueron mencionados en el Programa de Acción Específico de Investigación en Salud 2007-2012, pero no figuran en las estrategias y objetivos; en su lugar, se enfatizan primordialmente aspectos de infraestructura y administrativos. En el periodo se publicaron 145 artículos sobre DSS, cuyas temáticas más abordadas fueron "condiciones de salud", "sistemas de salud" y "nutrición y obesidad". Conclusiones. A pesar de que existe investigación en México sobre DSS, la instrumentación de esos hallazgos en políticas de salud no se ha implementado. El Programa Sectorial de Salud 2013-2018 representa una ventana de oportunidad para posicionar resultados de investigación que promuevan políticas de equidad en salud.
Objective. To examine the research on social determinants of health (SDH) produced in Mexico during the period 2005-2012, based on the characterization of the national health research system and the scientific production on this topic. Materials and methods. Two-stage analyses: Review of Mexican documents and official sources on health research and systematic bibliographic review of the literature on SDH. Results. Although SDH were mentioned in the Specific Action Plan for Health Research 2007-2012, they are not implemented in strategies and goals, as the emphasis is put mostly in infrastructure and administrative aspects of research. In the period studied, 145 articles were published on SDH topics such as health conditions, health systems and nutrition and obesity. Conclusions. In spite of the availability of research on SDH in Mexico, the operationalization of such findings into health policies has not been possible. The current Sectorial Program on Health 2013-2018 represents a window of opportunity to position research findings that promote health equity policies.
Subject(s)
Animals , Drosophila Proteins , Drosophila/physiology , Gene Expression Regulation, Developmental , Genes, Tumor Suppressor , Insect Hormones/genetics , Neuromuscular Junction/physiology , Synapses/physiology , Synapses/ultrastructure , Tumor Suppressor Proteins , Axons , Drosophila/genetics , Evoked Potentials , Genes, Insect , Insect Hormones/biosynthesis , Microscopy, Electron , Motor Neurons/physiology , Motor Neurons/ultrastructure , Muscles/innervation , Mutagenesis , Neuromuscular Junction/ultrastructure , Synaptic TransmissionABSTRACT
BACKGROUND: Cell-cell interactions are a basic principle for the organization of tissues and organs allowing them to perform integrated functions and to organize themselves spatially and temporally. Peptidic molecules secreted by neurons and epithelial cells play fundamental roles in cell-cell interactions, acting as local neuromodulators, neurohormones, as well as endocrine and paracrine messengers. Allatotropin (AT) is a neuropeptide originally described as a regulator of Juvenile Hormone synthesis, which plays multiple neural, endocrine and myoactive roles in insects and other organisms. METHODS: A combination of immunohistochemistry using AT-antibodies and AT-Qdot nanocrystal conjugates was used to identify immunoreactive nerve cells containing the peptide and epithelial-muscular cells targeted by AT in Hydra plagiodesmica. Physiological assays using AT and AT- antibodies revealed that while AT stimulated the extrusion of the hypostome in a dose-response fashion in starved hydroids, the activity of hypostome in hydroids challenged with food was blocked by treatments with different doses of AT-antibodies. CONCLUSIONS: AT antibodies immunolabeled nerve cells in the stalk, pedal disc, tentacles and hypostome. AT-Qdot conjugates recognized epithelial-muscular cell in the same tissues, suggesting the existence of anatomical and functional relationships between these two cell populations. Physiological assays indicated that the AT-like peptide is facilitating food ingestion. SIGNIFICANCE: Immunochemical, physiological and bioinformatics evidence advocates that AT is an ancestral neuropeptide involved in myoregulatory activities associated with meal ingestion and digestion.
Subject(s)
Feeding Behavior , Insect Hormones/metabolism , Neuropeptides/metabolism , Animals , Computational Biology , Gene Expression Regulation , Hydra/metabolism , Insect Hormones/chemistry , Neuropeptides/chemistry , Quantum Dots , Receptors, Neuropeptide/metabolismABSTRACT
Mechanisms coordinating cell-cell interaction have appeared early in evolution. Allatotropin (AT), a neuropeptide isolated based on its ability to stimulate the synthesis of juvenile hormones (JHs) in insects has also been found in other invertebrate phyla. Despite this function, AT has proved to be myotropic. In the present study we analyze its expression in two groups of Turbellaria (Catenulida, Macrostomida), and its probable relationship with muscle tissue. The results show the presence of an AT-like peptide in the free living turbellaria analyzed. The analysis of the expression of the peptide together with phalloidin, suggests a functional relationship between the peptide and muscle tissue, showing that it could be acting as a myoregulator. The finding of immunoreactive fibers associated with sensory organs like ciliated pits in Catenulida and eyes in Macrostomida makes probable that AT could play a role in the physiological mechanisms controlling circadian activities. Furthermore, the existence of AT in several phyla of Protostomata suggests that this peptide could be a synapomorphic feature of this group. Indeed, the presence in organisms that do not undergo metamorphosis, could be signaling that it was first involved in myotropic activities, being the stimulation of the synthesis of JHs a secondary function acquired by the phylum Arthropoda.
Subject(s)
Insect Hormones/metabolism , Muscles/metabolism , Neuropeptides/metabolism , Phalloidine/metabolism , Turbellaria/metabolism , Animals , Gene Expression Regulation , Insect Hormones/genetics , Juvenile Hormones/metabolism , Muscles/physiology , Neuropeptides/genetics , Phalloidine/genetics , Turbellaria/cytology , Turbellaria/geneticsABSTRACT
We show a straightforward workflow combining homology search in Rhodnius prolixus genome sequence with cloning by rapid amplification of cDNA ends and mass spectrometry. We have identified 32 genes and their transcripts that encode a number of neuropeptide precursors leading to 194 putative peptides. We validated by mass spectrometry 82 of those predicted neuropeptides in the brain of R. prolixus to achieve the first comprehensive genomic, transcriptomic and neuropeptidomic analysis of an insect disease vector. Comparisons of available insect neuropeptide sequences revealed that the R. prolixus genome contains most of the conserved neuropeptides in insects, many of them displaying specific features at the sequence level. Some gene families reported here are identified for the first time in the order Hemiptera, a highly biodiverse group of insects that includes many human, animal and plant disease agents.
Subject(s)
Insect Hormones/genetics , Neuropeptides/genetics , Protein Precursors/genetics , Rhodnius/genetics , Amino Acid Sequence , Animals , Brain Chemistry , Chagas Disease/transmission , Female , Genome, Insect , Insect Hormones/analysis , Insect Proteins/genetics , Insect Vectors/genetics , Male , Mass Spectrometry , Molecular Sequence Data , Multigene Family , Neuropeptides/analysis , Neuropeptides/classification , Protein Precursors/analysis , Rhodnius/chemistryABSTRACT
Allatotropin (AT) is a neuropeptide originally isolated from the brain of Manduca sexta and then characterized in several insect species. It acts as a neurohormone, as well as a neuromodulator. While it was primarily characterized on the basis of its ability to stimulate the secretion of juvenile hormones, it was also found that it acts as a cardioaccelerator and myostimulator. The presence of AT in IV instar larvae of T. infestans was previously described at the level of the Malpighian tubules. In the present study we report the presence of the peptide at the level of the brain, retrocerebral complex, as well as in the anterior midgut and aorta. The presence of AT at the corpora allata suggests that the peptide is acting on the gland during the first days of the moulting cycle. Neural processes at the level of the aorta and the anterior midgut suggests that, like in adults, the hormone is acting as a cardioaccelerator and myostimulator. The peptide was also found in open-type cells of the midgut. Finally the presence of allatotropic neurons in the optical lobe of the brain suggests that as in other species, the peptide is related with the control of circadian rhythms.
Subject(s)
Insect Hormones/metabolism , Neuropeptides/metabolism , Triatoma/metabolism , Animals , Brain/metabolism , Corpora Allata/metabolism , Gastrointestinal Tract/cytology , Gastrointestinal Tract/metabolism , Immunohistochemistry , Juvenile Hormones/metabolism , Larva/metabolism , Malpighian Tubules/metabolism , Manduca , Nervous System/cytology , Nervous System/metabolism , Neurons/cytology , Neurons/metabolism , Optic Lobe, Nonmammalian/cytology , Optic Lobe, Nonmammalian/metabolism , Triatoma/cytologyABSTRACT
Haematophagous insects incorporate a large quantity of blood with each meal, producing a big quantity of urine in a few hours. The activity of the Malpighian tubules (MTs) is facilitated by the increase of the circulation of the haemolymph produced by the increase of the aorta contractions as well as, of the peristaltic waves of the anterior midgut. MTs of Triatoma infestans secrete an allatotropin-like peptide, which has a myostimulatory effect on the hindgut, inducing the mixing and voiding of the content during post-prandial diuresis. We are reporting now the activity of allatotropin (AT) as a cardioacceleratory and a myostimulatory peptide at the level of the anterior midgut. The peptide induced the increase of the rate of contractions of the anterior midgut and the aorta in a wide range of concentrations. The cardioacceleratory effect of AT was dependent on the feeding state of the insects and on the presence of serotonin. The response showed the existence of a differential behavior between sexes, inducing a higher increase on the frequency of contractions, as well as, the width of the aorta in males than in females. Finally, our results suggest that AT interacts with serotonin to facilitate the circulation of haemolymph during post-prandial diuresis.