Red Pigment-concentrating Hormone Research Articles

Examination of the effects of triacylglycerol lipase on astaxanthin conversion in the black tiger prawn Penaeus monodon

The enzyme triacylglycerol lipase (TGL) catalyses the hydrolysis of triacylglycerols, and is characterized by catalytic esterification and ester exchange functionality. However, at present, there is comparatively little information regarding the sequences and functional validation of TGLs in crustaceans. In this study, we cloned the TGL1 gene of the black tiger prawn Penaeus monodon. The full-length cDNA of PmTGL1 comprises 1807 base pairs (bp), including an 80-bp 5′-untranslated region (UTR), a 149-bp 3′-UTR, and a 1578-bp open-reading frame (ORF), and encodes a putative 525-amino acid protein of 58.71 kDa with a theoretical isoelectric point of 4.81. Homology and phylogenetic tree analyses revealed that PmTGL1 is most closely related to the TGL of Penaeus chinensis, with which it clusters in a single group (similarity of up to 97.52%). Quantitative real-time PCR analysis further revealed that PmTGL1 is most highly expressed in the shrimp hepatopancreas. Although the expression of PmTGL1 was not significantly influenced by the differences in the background colour of the rearing pond, it was significantly promoted in response to dietary supplementation with astaxanthin. As an adipokinetic hormone in P. monodon, PmRPCH (red pigment-concentrating hormone) plays an important role in body colour changes and lipid metabolism. Based on our findings, we speculate that the PmTGL1 facilitates astaxanthin metabolism in P. monodon via the hydrolysis of esterified astaxanthin to yield free astaxanthin and that the activity of TGL in these shrimps is regulated via an adipokinetic hormone cascade.

APGW/AKH Precursor from Rotifer Brachionus plicatilis and the DNA Loss Model Explain Evolutionary Trends of the Neuropeptide LWamide, APGWamide, RPCH, AKH, ACP, CRZ, and GnRH Families

In the year 2002, DNA loss model (DNA-LM) postulated that neuropeptide genes to emerged through codons loss via the repair of damaged DNA from ancestral gene namely Neuropeptide Precursor Predictive (NPP), which organization correspond two or more neuropeptides precursors evolutive related. The DNA-LM was elaborated according to amino acids homology among LWamide, APGWamide, red pigment-concentrating hormone (RPCH), adipokinetic hormones (AKHs) and in silico APGW/RPCH NPPAPGW/AKH NPP were proposed. With the above principle, it was proposed the evolution of corazonin (CRZ), gonadotropin-releasing hormone (GnRH), AKH, and AKH/CRZ (ACP), but any NPP never was considered. However, the evolutive relation via DNA-LM among these neuropeptides precursors not has been established yet. Therefore, the transcriptomes from crabs Callinectes toxotes and Callinectes arcuatus were used to characterized ACP and partial CRZ precursors, respectively. BLAST alignment with APGW/RPCH NPP and APGW/AKH NPP allow identified similar NPP in the rotifer Brachionus plicatilis and other invertebrates. Moreover, three bioinformatics algorithms and manual verification were used to purify 13,778 sequences, generating a database with 719 neuropeptide precursors. Phylogenetic trees with the DNA-LM parameters showed that some ACP, CRZ, AKH2 and two NPP share nodes with GnRH from vertebrates and some of this neuropeptide had nodes in invertebrates. Whereas the phylogenetic tree with standard parameters do not showed previous node pattern. Robinson-Foulds metric corroborates the differences among phylogenetic trees. Homology relationship showed four putative orthogroups; AKH4, CRZ, and protostomes GnRH had individual group. This is the first demonstration of NPP in species and would explain the evolution neuropeptide families by the DNA-LM.

Modulation by Neuropeptides with Overlapping Targets Results in Functional Overlap in Oscillatory Circuit Activation.

Neuromodulation lends flexibility to neural circuit operation but the general notion that different neuromodulators sculpt neural circuit activity into distinct and characteristic patterns is complicated by interindividual variability. In addition, some neuromodulators converge onto the same signaling pathways, with similar effects on neurons and synapses. We compared the effects of three neuropeptides on the rhythmic pyloric circuit in the stomatogastric ganglion of male crabs, Cancer borealis Proctolin (PROC), crustacean cardioactive peptide (CCAP), and red pigment concentrating hormone (RPCH) activate the same modulatory inward current, I MI, and have convergent actions on synapses. However, while PROC targets all four neuron types in the core pyloric circuit, CCAP and RPCH target the same subset of only two neurons. After removal of spontaneous neuromodulator release, none of the neuropeptides restored the control cycle frequency, but all restored the relative timing between neuron types. Consequently, differences between neuropeptide effects were mainly found in the spiking activity of different neuron types. We performed statistical comparisons using the Euclidean distance in the multidimensional space of normalized output attributes to obtain a single measure of difference between modulatory states. Across preparations, the circuit output in PROC was distinguishable from CCAP and RPCH, but CCAP and RPCH were not distinguishable from each other. However, we argue that even between PROC and the other two neuropeptides, population data overlapped enough to prevent reliable identification of individual output patterns as characteristic for a specific neuropeptide. We confirmed this notion by showing that blind classifications by machine learning algorithms were only moderately successful.Significance Statement It is commonly assumed that distinct behaviors or circuit activities can be elicited by different neuromodulators. Yet it is unknown to what extent these characteristic actions remain distinct across individuals. We use a well-studied circuit model of neuromodulation to examine the effects of three neuropeptides, each known to produce a distinct activity pattern in controlled studies. We find that, when compared across individuals, the three peptides elicit activity patterns that are either statistically indistinguishable or show too much overlap to be labeled characteristic. We ascribe this to interindividual variability and overlapping subcellular actions of the modulators. Because both factors are common in all neural circuits, these findings have broad significance for understanding chemical neuromodulatory actions while considering interindividual variability.

Erratum: Eyestalk Ablation to Increase Ovarian Maturation in Mud Crabs.

Erratum: Eyestalk Ablation to Increase Ovarian Maturation in Mud Crabs.

Corrections.

In the presence of descending modulatory inputs, the stomatogastric ganglion (STG) of the lobster <i>Homarus americanus</i>generates a triphasic motor pattern, the pyloric rhythm. Red pigment-concentrating hormone (RPCH) and <i>Cancer borealis</i>tachykinin-related peptide (CabTRP) are colocalized in a pair of fibers that project into the neuropil of the STG. When the STG was isolated from anterior ganglia modulatory inputs, the lateral pyloric (LP) and pyloric (PY) neurons became silent, whereas the anterior burster (AB) and pyloric dilator (PD) neurons were rhythmically active at a low frequency. Exogenous application of 10⁻⁶m RPCH activated the LP neuron but not the PY neurons; 10⁻⁶m CabTRP activated the PY neurons but not the LP neuron. The actions of RPCH on the LP neuron and CabTRP on the PY neurons persisted when the rhythmic drive from the PD and AB neurons was removed, suggesting that the LP and PY neurons are direct targets for RPCH and CabTRP respectively. Coapplication of 10⁻⁶m RPCH and 10⁻⁶m CabTRP elicited triphasic motor patterns with phase relationships resembling those in a preparation with modulatory inputs intact. In summary, cotransmitters acting on different network targets act cooperatively to activate a complete central pattern-generating circuit.

Modulatory capacity correlates with dietary diversity in three species of decapod crustaceans

The central nervous systems of decapod crustaceans contains numerous amines, amino acids, and peptides that modulate the central pattern generating (CPG) circuits found within its stomatogastric nervous system (STNS) and cardiac ganglion (CG); over 30 modulators have been identified in one species of crab (C. borealis). CPGs in the STNS are responsible for internally chewing (Gastric Mill CPG) and filtering food (Pyloric CPG) into the midgut, while the CG drives the heartbeat. Neuromodulation allows these CPGs to flexibly produce a variety of output patterns; however, the variety of modulators exceeds the variety of induced output, leading us to ask why so many modulators are present. Although modulatory capacity, a measure of the extent to which a neuronal circuit can be modulated, may be solely an inherited trait, we hypothesize a functional correlation: decapods that need more behavioral flexibility utilize a broader range of modulators than those with more stereotyped behaviors. We thus predicted that decapods with broad diets would have a larger modulatory capacity in the STNS, but not the CG, than those with limited diets. To test this, we compared responses of the STNS and CG CPGs in three species of majoid crabs (Pugettia producta, Libinia emarginata, and Chionoecetes opilio) to the same neuromodulators. L. emarginata and C. opilio both have broad diets, while P. producta primarily eats kelp. With one exception, each species’ CG exhibited similar responses to all modulators. The STNS of crabs with broader diets responded similarly to all modulators, whereas the P. producta STNS showed limited responses to the same set of neuromodulators. CG responses did not follow this trend, as all crabs showed varied CG response to modulation. Modulators: Cancer borealisTachykinin Related Peptide (CabTRP), Crustacean Cardioactive Peptide (CCAP), Red Pigment Concentrating Hormone (RPCH), Myosuppressin (Myo), Oxotremorine (Oxo), Dopamine (DA), Proctolin (Proc).

Evidences for Red Pigment Concentrating Hormone (RPCH) and Beta-Pigment Dispersing Hormone (β-PDH) Inducing Oocyte Meiotic Maturation in the Chinese Mitten Crab, Eriocheir sinensis.

Red pigment concentrating hormone (RPCH) and pigment dispersing hormone (PDH) are crustacean neuropeptides involved in broad physiological processes including body color changes, circadian rhythm, and ovarian growth. In this study, the full-length cDNA of RPCH and PDH were identified from the brain of the Chinese mitten crab Eriocheir sinensis. The deduced RPCH and PDH mature peptides shared identical sequence to the adipokinetic hormone/RPCH peptides family and the β-PDH isoforms and were designated as Es-RPCH and Es-β-PDH, respectively. Es-RPCH and Es-β-PDH transcripts were distributed in the brain and eyestalks. The positive signals of Es-RPCH and Es-β-PDH were localized in the neuronal clusters 6, 8, 9, 10, and 17 of the brain as revealed by in situ hybridization. The expression level of Es-RPCH and Es-β-PDH mRNA in nervous tissues were all significantly increased at vitellogenic stage, and then decreased at the final meiotic maturation stage. The administrated with synthesized Es-RPCH peptide results in germinal vesicles shift toward the plasma membrane in vitellogenic oocyte, and significant decrease of the gonad-somatic index (GSI) and mean oocyte diameter as well as the expression of vitellogenin mRNA at 30 days post injection in vivo. Similar results were also found when injection of the Es-β-PDH peptide. In vitro culture demonstrated that Es-RPCH and Es-β-PDH induced germinal vesicle breakdown of the late vitellogenic oocytes. Comparative ovarian transcriptome analysis indicated that some reproduction/meiosis-related genes such as cdc2 kinase, cyclin B, 5-HT-R and retinoid-X receptor were significantly upregulated in response to Es-RPCH and Es-β-PDH treatments. Taken together, these results provided the evidence for the inductive effect of Es-RPCH and Es-β-PDH on the oocyte meiotic maturation in E. sinensis.

Roles of Neurotransmitters in Decapod Reproduction

In decapods neurotransmitters govern a variety of physiological events including growth, moulting, reproduction, locomotor activity, behavior and sexual differentiation. The most important reproductive neurotransmitters in decapods are histamine (HA), dopamine (DA), 5-hydrtryptamine (5HT, serotonin), leucine enkephalin (Leu-ENK), octopamine (OA), epinephrine (EPI), methionine enkephalin (Met-ENK), and Red pigment-concentrating hormone (RPCH). In this study, the discovery and identification of reproductive neurotransmitters in decapods are reviewed. The role of neurotransmitters (i.e. 5-HT, DA, OA, RPCH, Leu-ENK and Met-ENK) on the reproduction and sexual maturation of decapods are remarkable. It can be concluded that understanding the roles and effects of neurotransmitters are useful for further basic studies in reproductive biology of decapodsas well as commercial profitability in terms of development of new techniques for assisted reproduction in aquaculture.

Advances in the Science of Crustacean Reproductive Physiology and Potential Applications to New Seed Production Technology

Wilder, M.N., 2019. Advances in the science of crustacean reproductive physiology and potential applications to new seed production technology. In: Jithendran, K.P.; Saraswathy, R.; Balasubramanian, C.P.; Kumaraguru Vasagam, K.P.; Jayasankar, V.; Raghavan, R.; Alavandi, S.V., and Vijayan, K.K. (eds.), BRAQCON 2019: World Brackishwater Aquaculture Conference. Journal of Coastal Research, Special Issue No. 86, pp. 6–10. Coconut Creek (Florida), ISSN 0749-0208.The shrimp culture industry continues to show expansion world-wide, and a large number of crustacean species having commercial importance are found within the order Decapoda. At present, eyestalk ablation is frequently used in hatcheries for marine shrimp in order to induce maturation under captivity, but the method is considered undesirable from the viewpoint of animal welfare. In this regard, this review details the current status of knowledge regarding crustacean reproductive mechanisms, especially in relation to research covering vitellogenin structure, structure and function of vitellogenesis-inhibiting hormone (VIH), identity of putative factors stimulating vitellogenesis, and applications of such research to developing new seed production technology. Regarding vitellogenin, full structure is now known for nearly 20 decapod species, not only for prawns/shrimps, but also for certain crab and crayfish species; vitellogenin molecules in these species have a primary structure consisting of more than 2,500 amino acid residues. VIH is a member of the crustacean hyperglycemic hormone (CHH)-family. Mature CHH-family peptides are typically comprised of 72-78 amino acids having six conserved cysteine residues. The complete identity of vitellogenesis-stimulating hormone (VSH) remains elusive, but eyestalk hormones such as red pigment-concentrating hormone (RPCH) may play a role in promoting ovarian maturation in certain species. The challenge for the future is to seek a viable alternative to eyestalk ablation based on hormonal manipulation and application of the above knowledge.

Graded Transmission without Action Potentials Sustains Rhythmic Activity in Some But Not All Modulators That Activate the Same Current

Neurons in the central pattern-generating circuits in the crustacean stomatogastric ganglion (STG) release neurotransmitter both as a graded function of presynaptic membrane potential that persists in TTX and in response to action potentials. In the STG of the male crab Cancer borealis, the modulators oxotremorine, C. borealis tachykinin-related peptide Ia (CabTRP1a), red pigment concentrating hormone (RPCH), proctolin, TNRNFLRFamide, and crustacean cardioactive peptide (CCAP) produce and sustain robust pyloric rhythms by activating the same modulatory current (I MI), albeit on different subsets of pyloric network targets. The muscarinic agonist oxotremorine, and the peptides CabTRP1a and RPCH elicited rhythmic triphasic intracellular alternating fluctuations of activity in the presence of TTX. Intracellular waveforms of pyloric neurons in oxotremorine and CabTRP1a in TTX were similar to those in the intact rhythm, and phase relationships among neurons were conserved. Although cycle frequency was conserved in oxotremorine and TTX, it was altered in CabTRP1a in the presence of TTX. Both rhythms were primarily driven by the pacemaker kernel consisting of the Anterior Burster and Pyloric Dilator neurons. In contrast, in TTX the circuit remained silent in proctolin, TNRNFLRFamide, and CCAP. These experiments show that graded synaptic transmission in the absence of voltage-gated Na+ current is sufficient to sustain rhythmic motor activity in some, but not other, modulatory conditions, even when each modulator activates the same ionic current. This further demonstrates that similar rhythmic motor patterns can be produced by qualitatively different mechanisms, one that depends on the activity of voltage-gated Na+ channels, and one that can persist in their absence.SIGNIFICANCE STATEMENT The pyloric rhythm of the crab stomatogastric ganglion depends both on spike-mediated and graded synaptic transmission. We activate the pyloric rhythm with a wide variety of different neuromodulators, all of which converge on the same voltage-dependent inward current. Interestingly, when action potentials and spike-mediated transmission are blocked using TTX, we find that the muscarinic agonist oxotremorine and the neuropeptide CabTRP1a sustain rhythmic alternations and appropriate phases of activity in the absence of action potentials. In contrast, TTX blocks rhythmic activity in the presence of other modulators. This demonstrates fundamental differences in the burst-generation mechanisms in different modulators that would not be suspected on the basis of their cellular actions at the level of the targeted current.

Effects of the adipokinetic hormone/red pigment-concentrating hormone (AKH/RPCH) family of peptides on MK-801-induced schizophrenia models.

The adipokinetic and red pigment-concentrating hormone (AKH/RPCH) family of peptides controls fat, carbohydrate, and protein metabolism in insects. In our previous study, we showed that AKH possesses antidepressant, anxiolytic, and analgesic effects, causes hyperlocomotion, and exerts neuroprotective effects and increased brain neurotrophic factors in mice. The aim of this study was to investigate the effects of Anax imperator AKH (Ani-AKH), Libellula auripennis AKH (Lia-AKH), and Phormia-Terra hypertrehalosemic hormone (Pht-HrTH) on MK-801-induced memory deterioration in the active allothetic place avoidance test (AAPA) and MK-801-induced sensorimotor gating deficit in the prepulse inhibition test (PPI). In the AAPA task, Long-Evans rats were treated with Ani-AKH (2mg/kg), Lia-AKH (2mg/kg), Pht-HrTH (2mg/kg), MK-801 (0.15mg/kg), and the combination of MK-801 with the hormones subchronically. In the prepulse inhibition test, Wistar albino rats were treated with Ani-AKH (1mg/kg), Lia-AKH (1mg/kg), Pht-HrTH (1mg/kg), MK-801 (0.1mg/kg), or the combination of MK-801 with hormones acutely before the test. In our study, Ani-AKH (2mg/kg), Lia-AKH (2mg/kg), and Pht-HrTH (2mg/kg) reversed MK-801 (0.15mg/kg)-induced cognitive memory impairment effects in the AAPA task. Lia-AKH (1mg/kg) significantly potentiated the MK-801-induced PPI disruption, while Ani-AKH (1mg/kg) partially potentiated the impairment caused by MK-801, and Pht-HrTH did not modify the effect of MK-801. In conclusion, AKH had no effect in sensorimotor gating deficits in the PPI test in schizophrenia model while AKH improved memory in the schizophrenia model of MK-801.

Molecular cloning of red pigment-concentrating hormone (RPCH) from eyestalks of the whiteleg shrimp (Litopenaeus vannamei): Evaluation of the effects of the hormone on ovarian growth and the influence of serotonin (5-HT) on its expression

A cDNA (Liv-RPCH) encoding a putative red pigment-concentrating hormone was cloned from whiteleg shrimp (Litopenaeus vannamei) eyestalks. The deduced amino acid sequence of this cDNA was 93 residues long and consisted of a 21-residue signal peptide, a 9-residue red pigment-concentrating hormone (RPCH) mature peptide, a dibasic cleavage site, and a 61-residue RPCH precursor-related peptide (RPRP). The Liv-RPCH transcript was detected not only in neural tissues (eyestalks, brain, thoracic ganglia, and abdominal ganglia) but also in non-neural tissues (ovaries and heart). In vivo injection of synthetic RPCH (pELNFSPGW-NH2) caused significant increases in hemolymph vitellogenin (Vg) concentrations, ovarian Vg mRNA levels, and mean oocyte area, suggesting that RPCH has a positive role in promoting ovarian growth in L. vannamei. The effects of 5-HT on the expression of Liv-RPCH transcripts in selected tissues (eyestalks, brain, thoracic ganglia, and ovaries) were examined by using quantitative real-time PCR. Injection of 5-HT (20 μg g−1 body weight) resulted in elevated Liv-RPCH transcript levels in the ovaries 3 and 6 h after injection, but no significant effects were observed in the eyestalks, brain, or thoracic ganglia.

Functional Characterization and Signaling Systems of Corazonin and Red Pigment Concentrating Hormone in the Green Shore Crab, Carcinus maenas.

Neuropeptides play a central role as neurotransmitters, neuromodulators and hormones in orchestrating arthropod physiology. The post-genomic surge in identified neuropeptides and their putative receptors has not been matched by functional characterization of ligand-receptor pairs. Indeed, until very recently no G protein-coupled receptors (GPCRs) had been functionally defined in any crustacean. Here we explore the structurally-related, functionally-diverse gonadotropin-releasing hormone paralogs, corazonin (CRZ) and red-pigment concentrating hormone (RPCH) and their G-protein coupled receptors (GPCRs) in the crab, Carcinus maenas. Using aequorin luminescence to measure in vitro Ca2+ mobilization we demonstrated receptor-ligand pairings of CRZ and RPCH. CRZR-activated cell signaling in a dose-dependent manner (EC50 0.75 nM) and comparative studies with insect CRZ peptides suggest that the C-terminus of this peptide is important in receptor-ligand interaction. RPCH interacted with RPCHR with extremely high sensitivity (EC50 20 pM). Neither receptor bound GnRH, nor the AKH/CRZ-related peptide. Transcript distributions of both receptors indicate that CRZR expression was, unexpectedly, restricted to the Y-organs (YO). Application of CRZ peptide to YO had no effect on ecdysteroid biosynthesis, excepting a modest stimulation in early post-molt. CRZ had no effect on heart activity, blood glucose levels, lipid mobilization or pigment distribution in chromatophores, a scenario that reflected the distribution of its mRNA. Apart from the well-known activity of RPCH as a chromatophorotropin, it also indirectly elicited hyperglycemia (which was eyestalk-dependent). RPCHR mRNA was also expressed in the ovary, indicating possible roles in reproduction. The anatomy of CRZ and RPCH neurons in the nervous system is described in detail by immunohistochemistry and in situ hybridization. Each peptide has extensive but non-overlapping distribution in the CNS, and neuroanatomy suggests that both are possibly released from the post-commissural organs. This study is one of the first to deorphanize a GPCR in a crustacean and to provide evidence for hitherto unknown and diverse functions of these evolutionarily-related neuropeptides.

Interaction of the red pigment-concentrating hormone of the crustacean Daphnia pulex, with its cognate receptor, Dappu-RPCHR: A nuclear magnetic resonance and modeling study

The primary sequence of the red pigment-concentrating hormone (RPCH) receptor of the water flea, Daphnia pulex, was used in homology modeling to construct the first 3D model of a crustacean G-protein coupled receptor, Dappu-RPCHR. This receptor was found to belong to the class A subfamily of GPCRs with a disulfide bridge between Cys72 and Cys150 and an ionic lock between Arg97 and Thr224 and Thr220. NMR restrained molecular dynamics was used to determine the structure of an agonist, Dappu-RPCH, in a membrane-mimicking environment. The agonist was found to be flexible but has two main conformations in solution, both having β-turns. Docking of the predominant structure was used to find a binding pocket on the receptor. The pocket’s spatial location was similar to that of the AKH receptor of Anopheles gambiae. The binding affinity was −69kcalmol−1 with the N-terminus of Dappu-RPCH inserted between helices 4 and 6, and the C-terminus interacting with extra-cellular loop, ECL2. Upon binding, H-bonding to the peptide may activate the receptor. This development of the first Dappu-RPCH/Dappu-RPCHR model could be useful for understanding ligand-receptor interactions in crustaceans.

Characterisation and pharmacological analysis of a crustacean G protein-coupled receptor: the red pigment-concentrating hormone receptor of Daphnia pulex

This is the first pharmacological characterisation of a neuropeptide G protein-coupled receptor (GPCR) in a crustacean. We cloned the ORF of the red pigment-concentrating hormone from a German strain of Daphnia pulex (Dappu-RPCH), as well as that of the cognate receptor (Dappu-RPCHR). Dappu-RPCHR has the hallmarks of the rhodopsin superfamily of GPCRs, and is more similar to insect adipokinetic hormone (AKH) receptor sequences than to receptor sequences for AKH/corazonin-like peptide or corazonin. We provide experimental evidence that Dappu-RPCH specifically activates the receptor (EC50 value of 65 pM) in a mammalian cell-based bioluminescence assay. We further characterised the properties of the ligands for the Dappu-RPCHR by investigating the activities of a variety of naturally-occurring peptides (insect AKH and crustacean RPCH peptides). The insect AKHs had lower EC50 values than the crustacean RPCHs. In addition, we tested a series of Dappu-RPCH analogues, where one residue at a time is systematically replaced by an alanine to learn about the relative importance of the termini and side chains for activation. Mainly amino acids in positions 1 to 4 and 8 of Dappu-RPCH appear responsible for effective activation of Dappu-RPCHR. The substitution of Phe4 in Dappu-RPCH had the most damaging effect on its agonistic activity.

Characterization of the neuropeptidome of a Southern Ocean decapod, the Antarctic shrimp Chorismus antarcticus: Focusing on a new decapod ITP-like peptide belonging to the CHH peptide family

As part of the study of the resilience of Antarctic crustaceans to global warming, the shrimp Chorismus antarcticus was subjected to an analysis of global approach using the Next Generation Sequencing Illumina Hi-Seq platform. With this data a detailed study into the principal neuropeptides and neurohormones of this species have been undertaken. Total RNAs from whole animals were enriched with eyestalk extracts to ensure maximum sequencing depth of the different neurohormones and neuropeptides mainly expressed into the X organ-sinus gland complex, which is a major endocrine organ of their synthesis. Apart from the information that can provide the availability of the transcriptome of a polar crustacean, the study of neuropeptides of a caridean shrimp will partially fill the limited data available for this taxon. Illumina sequencing was used to produce a transcriptome of the polar shrimp. Analysis of the Trinity assembled contigs produced 55 pre-pro-peptides, coding for 111 neuropeptides belonging to the following families: adipokinetic-corazonin-like peptide, Allatostatins (A, B et C), Bursicon (α), CCHamide, Crustacean Hyperglycemic Hormones (CHH), Crustacean Cardioactive Peptide (CCAP), Corazonin, Crustacean Female Sex Hormone (CSFH), Diuretic Hormones 31 and 45 (DH), Eclosion Hormone (EH), FLRFamide, GSEFLamide, Intocin, Ion Transport Peptide-like (ITP-like), Leucokinin, Molt-inhibiting Hormone, Myosuppresin, Neuroparsin, Neuropeptide F (NPF), Orcokinin, Orcomyotropin, Pigment Dispersing Hormone (PDH), Pyrokinin, Red Pigment Concentrating Hormone (RPCH), SIFamide, small Neuropeptide F (sNPF), Sulfakinin and finally Tachykinin Related peptides. Among the new peptides highlighted in this study, the focus was placed on the peptides of the CHH family and more particularly on a new ITP-like in order to confirm its belonging to a new group of peptides of the family. A phylogeny made from more than 200 sequences of peptides, included new sequences from new species besides Chorismus antarcticus, confirms the peculiarity of this new set of peptides gathered under the name ITP-like.

Identification of genes that potentially affect social dominance hierarchy in adult male giant freshwater prawns (Macrobrachium rosenbergii)

The giant freshwater prawn (GFP) or Macrobrachium rosenbergii is one of the most important freshwater aquaculture crustaceans worldwide. An enduring problem with the improvement of this species in culture is the social dominance hierarchy that is manifested in male morphotypes where a few fast growing males (blue claw) individuals inhibit the growth of subdominant males. Here, we applied a transcriptomics approach to identify genes that contribute to expression of dominant phenotypes. Six different GFP tissues (abdominal muscle, hepatopancreas, stomach, claw, eyestalk and testis/ovary) were dissected for RNA extraction to prepare cDNA libraries for sequencing in Illumina NextSeq 500 platform. Illumina sequence reads were assembled and annotated for identifying candidate genes. This dataset currently provides the most comprehensive transcriptome data set available for GFP (147,507,623 reads assembled into 30,522 annotated contigs). 40 candidate genes were identified that could potentially play a role in determining adult male morphotype. Candidate genes identified include; Red pigment concentrating hormone (RPCH), Adipokinetic hormone receptor, Pigment dispersing hormone precursor (PDH), Pheromone and odorant, Visual pigment like receptor, Opsin, Glycoprotein 93 and Lola protein isoform a. The genes identified here provide a resource for understanding adult male morphotype development and the hierarchical dominance system in adult male GFP. In the future this information can be applied potentially to manipulate male morphotype frequencies in culture to bias adult male phenotypic ratios to improve overall productivity in practical genetic improvement programs.

Pigment Translocation in Caridean Shrimp Chromatophores: Receptor Type, Signal Transduction, Second Messengers, and Cross Talk Among Multiple Signaling Cascades.

Pigment aggregation in shrimp chromatophores is triggered by red pigment concentrating hormone (RPCH), a neurosecretory peptide whose plasma membrane receptor may be a G-protein coupled receptor (GPCR). While RPCH binding activates the Ca2+ /cGMP signaling cascades, a role for cyclic AMP (cAMP) in pigment aggregation is obscure, as are the steps governing Ca2+ release from the smooth endoplasmic reticulum (SER). A role for the antagonistic neuropeptide, pigment dispersing homone (α-PDH) is also unclear. In red, ovarian chromatophores from the freshwater shrimp Macrobrachium olfersi, we show that a G-protein antagonist (AntPG) strongly inhibits RPCH-triggered pigment aggregation, suggesting that RPCH binds to a GPCR, activating an inhibitory G-protein. Decreasing cAMP levels may cue pigment aggregation, since cytosolic cAMP titers, when augmented by cholera toxin, forskolin or vinpocentine, completely or partially impair pigment aggregation. Triggering opposing Ca2+ /cGMP and cAMP cascades by simultaneous perfusion with lipid-soluble cyclic nucleotide analogs induces a "tug-of-war" response, pigments aggregating in some chromatosomes with unpredictable, oscillatory movements in others. Inhibition of cAMP-dependent protein kinase accelerates aggregation and reduces dispersion velocities, suggesting a role in phosphorylation events, possibly regulating SER Ca2+ release and pigment aggregation. The second messengers IP3 and cADPR do not stimulate SER Ca2+ release. α-PDH does not sustain pigment dispersion, suggesting that pigment translocation in caridean chromatophores may be regulated solely by RPCH, since PDH is not required. We propose a working hypothesis to further unravel key steps in the mechanisms of pigment translocation within crustacean chromatophores that have remained obscure for nearly a century.

The decapod red pigment-concentrating hormone (Panbo-RPCH) is the first identified neuropeptide of the order Plecoptera and is interpreted as homoplastic character state

This paper presents the first neuropeptide structure, identified by mass spectrometry, in two species of Plectoptera (stoneflies) and in one species of the coleopteran family Lycidae. In all three species, the octapeptide Panbo-RPCH (first identified in Pandalus borealis as a red pigment-concentrating hormone: pGlu-Leu-Asn-Phe-Ser-Pro-Gly-Trp amide) is present. A review of the literature available on invertebrate neuropeptides that are identified or predicted from expressed sequence tags, transcriptome shotgun assemblies, and from fully sequenced genomes, show that Panbo-RPCH is found in Malacostraca (Crustacea) and certain hemipteran Heteroptera (Insecta). To date, Panbo-RPCH has not been shown present in non-Malacostracan crustaceans, nor in basal taxa of the Insecta (Archaeognatha, Zygentoma, Ephemeroptera, Odonata). The present data adds to knowledge on the distribution of Panbo-RPCH, and when taking into account the most accepted, current phylogenetics of the Crustacea–Hexapoda relationship, this distribution of Panbo-RPCH in Malacostraca, Plecoptera, some hemipteran Heteroptera and in Coleoptera (Lycidae) can best be explained by homoplasy, implying parallel evolution.

Localization and functional characterization of a novel adipokinetic hormone in the mollusk, Aplysia californica.

Increasing evidence suggests that gonadotropin-releasing hormone (GnRH), corazonin, adipokinetic hormone (AKH), and red pigment-concentrating hormone all share common ancestry to form a GnRH superfamily. Despite the wide presence of these peptides in protostomes, their biological effects remain poorly characterized in many taxa. This study had three goals. First, we cloned the full-length sequence of a novel AKH, termed Aplysia-AKH, and examined its distribution in an opisthobranch mollusk, Aplysia californica. Second, we investigated in vivo biological effects of Aplysia-AKH. Lastly, we compared the effects of Aplysia-AKH to a related A. californica peptide, Aplysia-GnRH. Results suggest that Aplysia-AKH mRNA and peptide are localized exclusively in central tissues, with abdominal, cerebral, and pleural ganglia being the primary sites of Aplysia-AKH production. However, Aplysia-AKH-positive fibers were found in all central ganglia, suggesting diverse neuromodulatory roles. Injections of A. californica with Aplysia-AKH significantly inhibited feeding, reduced body mass, increased excretion of feces, and reduced gonadal mass and oocyte diameter. The in vivo effects of Aplysia-AKH differed substantially from Aplysia-GnRH. Overall, the distribution and biological effects of Aplysia-AKH suggest it has diverged functionally from Aplysia-GnRH over the course of evolution. Further, that both Aplysia-AKH and Aplysia-GnRH failed to activate reproduction suggest the critical role of GnRH as a reproductive activator may be a phenomenon unique to vertebrates.