ABSTRACT
Venom systems are complex traits that have independently emerged multiple times in diverse plant and animal phyla. Within each venomous lineage there typically exists interspecific variation in venom composition where several factors have been proposed as drivers of variation, including phylogeny and diet. Understanding these factors is of broad biological interest and has implications for the development of antivenom therapies and venom-based drug discovery. Because of their high species richness and the presence of several major evolutionary prey shifts, venomous marine cone snails (genus Conus) provide an ideal system to investigate drivers of interspecific venom variation. Here, by analyzing the venom gland expression profiles of â¼3,000 toxin genes from 42 species of cone snail, we elucidate the role of prey-specific selection pressures in shaping venom variation. By analyzing overall venom composition and individual toxin structures, we demonstrate that the shifts from vermivory to piscivory in Conus are complemented by distinct changes in venom composition independent of phylogeny. In vivo injections of venom from piscivorous cone snails in fish further showed a higher potency compared with venom of nonpiscivores demonstrating a selective advantage. Together, our findings provide compelling evidence for the role of prey shifts in directing the venom composition of cone snails and expand our understanding of the mechanisms of venom variation and diversification.
Subject(s)
Conus Snail , Mollusk Venoms , Animals , Conus Snail/genetics , Mollusk Venoms/genetics , Predatory Behavior , Biological Evolution , Phylogeny , Evolution, MolecularABSTRACT
The venom of cone snails has been proven to be a rich source of bioactive peptides that target a variety of ion channels and receptors. α-Conotoxins (αCtx) interact with nicotinic acetylcholine receptors (nAChRs) and are powerful tools for investigating the structure and function of the various nAChR subtypes. By studying how conotoxins interact with nAChRs, we can improve our understanding of these receptors, leading to new insights into neurological diseases associated with nAChRs. Here, we describe the discovery and characterization of a novel conotoxin from Conus ateralbus, αCtx-AtIA, which has an amino acid sequence homologous to the well-described αCtx-PeIA, but with a different selectivity profile towards nAChRs. We tested the synthetic αCtx-AtIA using the calcium imaging-based Constellation Pharmacology assay on mouse DRG neurons and found that αCtx-AtIA significantly inhibited ACh-induced calcium influx in the presence of an α7 positive allosteric modulator, PNU-120596 (PNU). However, αCtx-AtIA did not display any activity in the absence of PNU. These findings were further validated using two-electrode voltage clamp electrophysiology performed on oocytes overexpressing mouse α3ß4, α6/α3ß4 and α7 nAChRs subtypes. We observed that αCtx-AtIA displayed no or low potency in blocking α3ß4 and α6/α3ß4 receptors, respectively, but improved potency and selectivity to block α7 nAChRs when compared with αCtx-PeIA. Through the synthesis of two additional analogs of αCtx-AtIA and subsequent characterization using Constellation Pharmacology, we were able to identify residue Trp18 as a major contributor to the activity of the peptide.
Subject(s)
Conotoxins , Conus Snail , Receptors, Nicotinic , Animals , Mice , Calcium , Amino Acid Sequence , alpha7 Nicotinic Acetylcholine ReceptorABSTRACT
The reaction of tris(pyridin-2-yl)amine with [CyRuCl2]2 (Cy = p-isopropyltoluene or cymene) in refluxing diglyme led to the formation of cis-[RuCl2{κ2-(2-py)3N}2]·CHCl3 (1a) after recrystallization from chloroform/pentane, or cis-dichloridobis[tris(pyridin-2-yl)amine-κ2N,N']ruthenium(II) dichloromethane disolvate, [RuCl2(C15H12N4)2]·2CH2Cl2 or cis-[RuCl2{κ2-(2-py)3N}2]·2CH2Cl2 (1b). Treatment of 1a with one equivalent of silver(I) hexafluoridoantimonate in dichloromethane gave [RuCl{κ2-(2-py)3N}{κ3-(2-py)3N}][SbF6]·CH2Cl2 (2a). Crystallization of 2a from chloroform provided chlorido[tris(pyridin-2-yl)amine-κ2N,N'][tris(pyridin-2-yl)amine-κ3N,N',N'']ruthenium(II) hexafluoridoantimonate chloroform monosolvate, [RuCl(C15H12N4)2][SbF6]·CHCl3 or [RuCl{κ2-(2-py)3N}{κ3-(2-py)3N}][SbF6]·CHCl3 (2b). Complex 2a reacted with a further equivalent of silver(I) hexafluoridoantimonate to give [Ru{κ3-(2-py)3N}2][SbF6]2 (3). The reaction of (2-py)3N with [CyRuCl2]2 in dichloromethane followed by treatment with excess sodium hexafluoridoantimonate gave the known complex [CyRuCl{κ2-(2-py)3N}][SbF6] (4). Complex 2 is a rare example of a complex containing both κ2- and κ3-(2-py)3N. Intramolecular π-stacking interactions determine the orientation of the free pyridyl in the κ2 complexes. An interesting encapsulation of methylene chloride hydrogen-bonded tetramers was noted in one case.
ABSTRACT
Peptide hormones and neuropeptides form a diverse class of bioactive secreted molecules that control essential processes in animals. Despite breakthroughs in peptide discovery, many signaling peptides remain undiscovered. Recently, we demonstrated the use of somatostatin-mimicking toxins from cone snails to identify the invertebrate ortholog of somatostatin. Here, we show that this toxin-based approach can be systematically applied to discover other unknown secretory peptides that are likely to have signaling function. Using large sequencing datasets, we searched for homologies between cone snail toxins and secreted proteins from the snails' prey. We identified and confirmed expression of five toxin families that share strong similarities with unknown secretory peptides from mollusks and annelids and in one case also from ecdysozoans. Based on several lines of evidence we propose that these peptides likely act as signaling peptides that serve important physiological functions. Indeed, we confirmed that one of the identified peptides belongs to the family of crustacean hyperglycemic hormone, a peptide not previously observed in Spiralia. We propose that this discovery pipeline can be broadly applied to other systems in which one organism has evolved molecules to manipulate the physiology of another.
ABSTRACT
Lymantria dispar L. is an invasive, non-native defoliating Lepidopteran established in North America that feeds on forest and urban trees. While many products are available to manage L. dispar post-emergence, few exist to prevent egg hatch when applied to egg masses. Here, we present the results of 3 separate experiments aimed at determining the efficacy of pre-emergent insecticides against L. dispar egg hatch. We found that the labeled rate (1:1) of Golden Pest Spray Oil (GPSO; AI: 93% soybean oil) can prevent L. dispar larvae from emerging in both field and lab assays. In large public spaces, we found that this treatment was ineffective at preventing L. dispar emergence or defoliation. Acelepryn (AI: 18.4% chlorantraniliprole) resulted in some suppression of egg hatch at a very low rate (.06 ml/ 3.8 liter) in both lab and field settings and the efficacy of higher rates should be further investigated. We also tested GPSO against Lepitect (97.4% acephate) in a public area that also received a Foray 48B (12.65% Bacillus thuringiensis, subsp. kurstaki) aerial application. On large oak trees in public areas, GPSO and Lepitect were not effective at reducing defoliation. Dormant pesticide applications generally reduce the risk of affecting negatively predator and parasitoid communities and are therefore desirable. Lymantria dispar pre-egg hatch applications will not work in every situation but should be considered as part of an integrated pest management (IPM) strategy for individual homeowner trees where thorough coverage can be obtained.
Subject(s)
Bacillus thuringiensis , Insecticides , Lepidoptera , Moths , Animals , Larva , Forests , TreesABSTRACT
Pain is often debilitating, and current treatments are neither universally efficacious nor without risks. Transient receptor potential (TRP) ion channels offer alternative targets for pain relief, but little is known about the regulation or identities of endogenous TRP ligands that affect inflammation and pain. Here, transcriptomic and targeted lipidomic analysis of damaged tissue from the mouse spinal nerve ligation (SNL)-induced chronic pain model revealed a time-dependent increase in Cyp1b1 mRNA and a concurrent accumulation of 8,9-epoxyeicosatrienoic acid (EET) and 19,20-EpDPA post injury. Production of 8,9-EET and 19,20-EpDPA by human/mouse CYP1B1 was confirmed in vitro, and 8,9-EET and 19,20-EpDPA selectively and dose-dependently sensitized and activated TRPA1 in overexpressing HEK-293 cells and Trpa1-expressing/AITC-responsive cultured mouse peptidergic dorsal root ganglia (DRG) neurons. TRPA1 activation by 8,9-EET and 19,20-EpDPA was attenuated by the antagonist A967079, and mouse TRPA1 was more responsive to 8,9-EET and 19,20-EpDPA than human TRPA1. This latter effect mapped to residues Y933, G939, and S921 of TRPA1. Intra-plantar injection of 19,20-EpDPA induced acute mechanical, but not thermal hypersensitivity in mice, which was also blocked by A967079. Similarly, Cyp1b1-knockout mice displayed a reduced chronic pain phenotype following SNL injury. These data suggest that manipulation of the CYP1B1-oxylipin-TRPA1 axis might have therapeutic benefit.
ABSTRACT
Somatostatin and its related peptides (SSRPs) form an important family of hormones with diverse physiological roles. The ubiquitous presence of SSRPs in vertebrates and several invertebrate deuterostomes suggests an ancient origin of the SSRP signaling system. However, the existence of SSRP genes outside of deuterostomes has not been established, and the evolutionary history of this signaling system remains poorly understood. Our recent discovery of SSRP-like toxins (consomatins) in venomous marine cone snails (Conus) suggested the presence of a related signaling system in mollusks and potentially other protostomes. Here, we identify the molluscan SSRP-like signaling gene that gave rise to the consomatin family. Following recruitment into venom, consomatin genes experienced strong positive selection and repeated gene duplications resulting in the formation of a hyperdiverse family of venom peptides. Intriguingly, the largest number of consomatins was found in worm-hunting species (>400 sequences), indicating a homologous system in annelids, another large protostome phylum. Consistent with this, comprehensive sequence mining enabled the identification of SSRP-like sequences (and their corresponding orphan receptor) in annelids and several other protostome phyla. These results established the existence of SSRP-like peptides in many major branches of bilaterians and challenge the prevailing hypothesis that deuterostome SSRPs and protostome allatostatin-C are orthologous peptide families. Finally, having a large set of predator-prey SSRP sequences available, we show that although the cone snail's signaling SSRP-like genes are under purifying selection, the venom consomatin genes experience rapid directional selection to target receptors in a changing mix of prey.
Subject(s)
Conotoxins , Conus Snail , Animals , Conotoxins/genetics , Conus Snail/genetics , Neuropeptides , Peptides/genetics , Somatostatin/genetics , VenomsABSTRACT
The venomous marine snails are conventionally divided into three groups, the cone snails (family Conidae), the auger snails (family Terebridae) and the turrids (formerly all assigned to a single family, Turridae). In this study, a library of venom peptides from species conventionally assigned to the genus Turris was correlated to a phylogenetic analysis. Nucleotide sequences of multiple genes from transcriptomes were used to assess the phylogenetic relationships across a diverse set of species. The resulting tree shows that as conventionally defined, the conoidean genus Turris, is polyphyletic. We describe a new genus, Purpuraturris gen. nov., that comprises the outlier species. In addition to morphological distinctions, molecular data reveal that this group is divergent from Turris sensu stricto. The correlation between phylogenetic information and a family of peptide sequences was used to highlight those peptides mostly likely to be unique and intimately associated with biological diversity. The plethora of peptide sequences available requires two prioritization decisions: which subset of peptides to initially characterize, and after these are characterized, which to comprehensively investigate for potential biomedical applications such as drug developments. Life Science Identifiers: urn:lsid:zoobank.org; pub: 60D46561-28F0-4C39-BAC4-66DC8B4EAEA4.
ABSTRACT
Marine tunicates produce defensive amino-acid-derived metabolites, including 2-(3,5-diiodo-4-methoxyphenyl)ethan-1-amine (DIMTA), but their mechanisms of action are rarely known. Using an assay-guided approach, we found that out of the many different sensory cells in the mouse dorsal root ganglion (DRG), DIMTA selectively affected low-threshold cold thermosensors. Whole-cell electrophysiology experiments using DRG cells, channels expressed in Xenopus oocytes, and human cell lines revealed that DIMTA blocks several potassium channels, reducing the magnitude of the afterhyperpolarization and increasing the baseline intracellular calcium concentration [Ca2+]i of low-threshold cold thermosensors. When injected into mice, DIMTA increased the threshold of cold sensation by >3 °C. DIMTA may thus serve as a lead in the further design of compounds that inhibit problems in the cold-sensory system, such as cold allodynia and other neuropathic pain conditions.
Subject(s)
Amines/metabolism , Calcium Channels/metabolism , Sensory Receptor Cells/metabolism , Amines/administration & dosage , Animals , Calcium/metabolism , Ganglia, Spinal/metabolism , Male , Mice , Patch-Clamp Techniques , Signal Transduction , Thermosensing/physiology , Urochordata , VertebratesABSTRACT
In our efforts to discover new drugs to treat pain, we identified molleamines A-E (1-5) as major neuroactive components of the sea slug, Pleurobranchus forskalii, and their prey, Didemnum molle, tunicates. The chemical structures of molleamines were elucidated by spectroscopy and confirmed by the total synthesis of molleamines A (1) and C (3). Synthetic 3 completely blocked acetylcholine-induced calcium flux in peptidergic nociceptors (PNs) in the somatosensory nervous system. Compound 3 affected neither the α7 nAChR nor the muscarinic acetylcholine receptors in calcium flux assays. In addition to nociceptors, 3 partially blocked the acetylcholine-induced calcium flux in the sympathetic nervous system, including neurons from the superior cervical ganglion. Electrophysiology revealed a block of α3ß4 (mouse) and α6/α3ß4 (rat) nicotinic acetylcholine receptors (nAChRs), with IC50 values of 1.4 and 3.1 µM, respectively. Molleamine C (3) is a partial antagonist, reaching a maximum block of 76-82% of the acetylcholine signal and showing no partial agonist response. Molleamine C (3) may thus provide a lead compound for the development of neuroactive compounds with unique biological properties.
Subject(s)
Receptors, Nicotinic , Urochordata , Animals , Aplysia , Mice , Nicotinic Antagonists/pharmacology , Nylons , Rats , alpha7 Nicotinic Acetylcholine ReceptorABSTRACT
In a program to identify pain treatments with low addiction potential, we isolated five steroids, conosteroids A-E (1-5), from the hypobranchial gland of the mollusk Conus geographus. Compounds 1-5 were active in a mouse dorsal root ganglion (DRG) assay that suggested that they might be analgesic. A synthetic analogue 6 was used for a detailed pharmacological study. Compound 6 significantly increased the pain threshold in mice in the hot-plate test at 2 and 50 mg/kg. Compound 6 at 500 nM antagonizes type-A γ-aminobutyric acid receptors (GABAARs). In a patch-clamp experiment, out of the six subunit combinations tested, 6 exhibited subtype selectivity, most strongly antagonizing α1ß1γ2 and α4ß3γ2 receptors (IC50 1.5 and 1.0 µM, respectively). Although the structures of 1-6 differ from those of known neuroactive steroids, they are cell-type-selective modulators of GABAARs, expanding the known chemical space of neuroactive steroids.
Subject(s)
Analgesics/chemistry , Conus Snail/chemistry , GABA Antagonists/chemistry , Neurosteroids/chemistry , Receptors, GABA/chemistry , Action Potentials/drug effects , Analgesics/chemical synthesis , Analgesics/pharmacology , Analgesics/therapeutic use , Animals , Conus Snail/metabolism , Disease Models, Animal , GABA Antagonists/isolation & purification , GABA Antagonists/pharmacology , GABA Antagonists/therapeutic use , Ganglia, Spinal/cytology , Ganglia, Spinal/drug effects , Ganglia, Spinal/physiology , Mice , Mice, Inbred C57BL , Molecular Conformation , Neurosteroids/isolation & purification , Neurosteroids/pharmacology , Neurosteroids/therapeutic use , Pain/chemically induced , Pain/drug therapy , Pain/pathology , Protein Subunits/chemistry , Protein Subunits/metabolism , Receptors, GABA/metabolismABSTRACT
Natural products such as conotoxins have tremendous potential as tools for biomedical research and for the treatment of different human diseases. Conotoxins are peptides present in the venoms of predatory cone snails that have a rich diversity of pharmacological functions. One of the major bottlenecks in natural products research is the rapid identification and evaluation of bioactive molecules. To overcome this limitation, we designed a set of light-induced behavioral assays in zebrafish larvae to screen for bioactive conotoxins. We used this screening approach to test several unique conotoxins derived from different cone snail clades and discovered that a conorfamide from Conus episcopatus, CNF-Ep1, had the most dramatic alterations in the locomotor behavior of zebrafish larvae. Interestingly, CNF-Ep1 is also bioactive in several mouse assay systems when tested in vitro and in vivo. Our novel screening platform can thus accelerate the identification of bioactive marine natural products, and the first compound discovered using this assay has intriguing properties that may uncover novel neuronal circuitry.
Subject(s)
Larva/drug effects , Locomotion/drug effects , Mollusk Venoms/pharmacology , Neuropeptides/pharmacology , Zebrafish , Animals , Conus Snail/chemistry , Female , Male , MiceABSTRACT
Current drug discovery efforts focus on identifying lead compounds acting on a molecular target associated with an established pathological state. Concerted molecular changes that occur in specific cell types during disease progression have generally not been identified. Here, we used constellation pharmacology to investigate rat dorsal root ganglion neurons using two models of peripheral nerve injury: chronic constriction injury (CCI) and spinal nerve ligation (SNL). In these well-established models of neuropathic pain, we show that the onset of chronic pain is accompanied by a dramatic, previously unreported increase in the number of bradykinin-responsive neurons, with larger increases observed after SNL relative to CCI. To define the neurons with altered expression, we charted the temporal course of molecular changes following 1, 3, 6, and 14 d after SNL injury and demonstrated that specific molecular changes have different time courses during the progression to a pain state. In particular, ATP receptors up-regulated on day 1 postinjury, whereas the increase in bradykinin receptors was gradual after day 3 postinjury. We specifically tracked changes in two subsets of neurons: peptidergic and nonpeptidergic nociceptors. Significant increases occurred in ATP responses in nAChR-expressing isolectin B4+ nonpeptidergic neurons 1 d postinjury, whereas peptidergic neurons did not display any significant change. We propose that remodeling of ion channels and receptors occurs in a concerted and cell-specific manner, resulting in the appearance of bradykinin-responsive neuronal subclasses that are relevant to chronic pain.
Subject(s)
Neurons/metabolism , Peripheral Nerve Injuries/pathology , Somatosensory Cortex/metabolism , Animals , Ganglia, Spinal/metabolism , Hyperalgesia/metabolism , Male , Neuralgia/metabolism , Nociceptors/metabolism , Rats , Rats, Sprague-Dawley , Spinal Nerves/metabolismABSTRACT
Identification and characterization of neuronal cell classes in motor circuits are essential for understanding the neural basis of behavior. It is a challenging task, especially in a non-genetic-model organism, to identify cell-specific expression of functional macromolecules. Here, we performed constellation pharmacology, calcium imaging of dissociated neurons to pharmacologically identify functional receptors expressed by vocal neurons in adult male and female African clawed frogs, Xenopus laevis. Previously we identified a population of vocal neurons called fast trill neurons (FTNs) in the amphibian parabrachial nucleus (PB) that express N-methyl-d-aspartate (NMDA) receptors and GABA and/or glycine receptors. Using constellation pharmacology, we identified four cell classes of putative fast trill neurons (pFTNs, responsive to both NMDA and GABA/glycine applications). We discovered that some pFTNs responded to the application of substance P (SP), acetylcholine (ACh), or both. Electrophysiological recordings obtained from FTNs using an ex vivo preparation verified that SP and/or ACh depolarize FTNs. Bilateral injection of ACh, SP, or their antagonists into PBs showed that ACh receptors are not sufficient but necessary for vocal production, and SP receptors play a role in shaping the morphology of vocalizations. Additionally, we discovered that the PB of adult female X. laevis also contains all the subclasses of neurons at a similar frequency as in males, despite their sexually distinct vocalizations. These results reveal novel neuromodulators that regulate X. laevis vocal production and demonstrate the power of constellation pharmacology in identifying the neuronal subtypes marked by functional expression of cell-specific receptors in non-genetic-model organisms.NEW & NOTEWORTHY Molecular profiles of neurons are critical for understanding the neuronal functions, but their identification is challenging especially in non-genetic-model organisms. Here, we characterized the functional expression of membrane macromolecules in vocal neurons of African clawed frogs, Xenopus laevis, using a technique called constellation pharmacology. We discovered that receptors for acetylcholine and/or substance P are expressed by some classes of vocal neurons, and their activation plays a role in the production of normal vocalizations.
Subject(s)
Neurons/physiology , Neurotransmitter Agents/pharmacology , Parabrachial Nucleus/physiology , Receptors, Neurotransmitter/metabolism , Vocalization, Animal/physiology , Xenopus laevis/physiology , Animals , Cells, Cultured , Female , Glycine/metabolism , Male , Microscopy, Fluorescence , N-Methylaspartate/metabolism , Neurons/classification , Neurons/metabolism , Parabrachial Nucleus/metabolism , Patch-Clamp Techniques , Pharmacology/methods , Receptors, Neurotransmitter/agonists , Receptors, Neurotransmitter/antagonists & inhibitors , Substance P/metabolism , Xenopus laevis/metabolism , gamma-Aminobutyric Acid/metabolismABSTRACT
Somatosensory neurons have historically been classified by a variety of approaches, including structural, anatomical, and genetic markers; electrophysiological properties; pharmacological sensitivities; and more recently, transcriptional profile differentiation. These methodologies, used separately, have yielded inconsistent classification schemes. Here, we describe phenotypic differences in response to pharmacological agents as measured by changes in cytosolic calcium concentration for the rapid classification of neurons in vitro; further analysis with genetic markers, whole-cell recordings, and single-cell transcriptomics validated these findings in a functional context. Using this general approach, which we refer to as tripartite constellation analysis (TCA), we focused on large-diameter dorsal-root ganglion (L-DRG) neurons with myelinated axons. Divergent responses to the K-channel antagonist, κM-conopeptide RIIIJ (RIIIJ), reliably identified six discrete functional cell classes. In two neuronal subclasses (L1 and L2), block with RIIIJ led to an increase in [Ca] i Simultaneous electrophysiology and calcium imaging showed that the RIIIJ-elicited increase in [Ca] i corresponded to different patterns of action potentials (APs), a train of APs in L1 neurons, and sporadic firing in L2 neurons. Genetically labeled mice established that L1 neurons are proprioceptors. The single-cell transcriptomes of L1 and L2 neurons showed that L2 neurons are Aδ-low-threshold mechanoreceptors. RIIIJ effects were replicated by application of the Kv1.1 selective antagonist, Dendrotoxin-K, in several L-DRG subclasses (L1, L2, L3, and L5), suggesting the presence of functional Kv1.1/Kv1.2 heteromeric channels. Using this approach on other neuronal subclasses should ultimately accelerate the comprehensive classification and characterization of individual somatosensory neuronal subclasses within a mixed population.
Subject(s)
Ganglia, Spinal/cytology , Sensory Receptor Cells/classification , Sensory Receptor Cells/physiology , Animals , Calcium/metabolism , Conotoxins/pharmacology , Cytosol/metabolism , Ganglia, Spinal/drug effects , Kv1.1 Potassium Channel/antagonists & inhibitors , Mice , Mice, Transgenic , Peptides/pharmacology , Potassium Channel Blockers/pharmacology , Sensory Receptor Cells/drug effects , Single-Cell Analysis , TranscriptomeABSTRACT
Invasions by insects introduced via international trade continue to cause worldwide impacts. Surveillance programs using traps baited with host volatiles and pheromones can detect incursions of nonnative species. We report on two experiments executed to determine if attractants for several insect species can be combined without compromising trap catches and detection ability of target species. In the first experiment, we tested the effect of bark beetle pheromones (plus α-pinene) and trap contact with foliage on trap catches of the brown marmorated stink bug Halyomorpha halys Stål (Hemiptera: Pentatomidae) in traps baited with a mixture of bisabolenes and methyl (E,E,Z)-2,4,6-decatrienoate. Trap capture of H. halys adults was greater in traps not in contact with foliage, and the bark beetle pheromones ipsenol and ipsdienol did not affect trap capture of H. halys. In the second experiment, we tested the effects of multi-lure interactions among the primary host attractants α-pinene and ethanol, and the pheromones monochamol, ipsenol, ipsdienol, lanierone, and the H. halys compounds, on trap captures of various forest and agricultural insect pests. Specifically, we targeted Monochamus spp. (Coleoptera: Cerambycidae), Ips spp. (Coleoptera: Scolytinae) and H. halys. We found that a combination of all lures did not catch significantly lower numbers of Monochamus carolinensis Olivier, Monochamus scutellatus Say (Coleoptera: Cerambycidae), and Ips pini Say (Coleoptera: Scolytidae) than lure combinations missing components although removal of both lanierone and ipsdienol somewhat increased catches of Ips grandicollis Eichhoff (Coleoptera: Curculionidae). Our results support the use of traps baited with a full combination of these attractants in surveillance programs. This should reduce costs and increase detection rates of a wider range of conifer forest pests and H. halys.
Subject(s)
Coleoptera , Heteroptera , Insect Control/methods , Introduced Species , Pheromones , Animals , Female , MaleABSTRACT
The molecular sensor of innocuous (painless) cold sensation is well-established to be transient receptor potential cation channel, subfamily M, member 8 (TRPM8). However, the role of transient receptor potential cation channel, subfamily A, member 1 (TRPA1) in noxious (painful) cold sensation has been controversial. We find that TRPA1 channels contribute to the noxious cold sensitivity of mouse somatosensory neurons, independent of TRPM8 channels, and that TRPA1-expressing neurons are largely non-overlapping with TRPM8-expressing neurons in mouse dorsal-root ganglia (DRG). However, relatively few TRPA1-expressing neurons (e.g., responsive to allyl isothiocyanate or AITC, a selective TRPA1 agonist) respond overtly to cold temperature in vitro, unlike TRPM8-expressing neurons, which almost all respond to cold. Using somatosensory neurons from TRPM8-/- mice and subtype-selective blockers of TRPM8 and TRPA1 channels, we demonstrate that responses to cold temperatures from TRPA1-expressing neurons are mediated by TRPA1 channels. We also identify two factors that affect the cold-sensitivity of TRPA1-expressing neurons: (1) cold-sensitive AITC-sensitive neurons express relatively more TRPA1 transcripts than cold-insensitive AITC-sensitive neurons and (2) voltage-gated potassium (KV) channels attenuate the cold-sensitivity of some TRPA1-expressing neurons. The combination of these two factors, combined with the relatively weak agonist-like activity of cold temperature on TRPA1 channels, partially explains why few TRPA1-expressing neurons respond to cold. Blocking KV channels also reveals another subclass of noxious cold-sensitive DRG neurons that do not express TRPM8 or TRPA1 channels. Altogether, the results of this study provide novel insights into the cold-sensitivity of different subclasses of somatosensory neurons.
Subject(s)
Cold Temperature , Ganglia, Spinal/physiology , Neurons/physiology , Nociception/physiology , Potassium Channels, Voltage-Gated/physiology , TRPA1 Cation Channel/physiology , Thermoreceptors/physiology , Thermosensing , Animals , Cells, Cultured , Female , Ganglia, Spinal/drug effects , Ganglia, Spinal/metabolism , HEK293 Cells , Humans , Isothiocyanates/administration & dosage , Male , Menthol/administration & dosage , Mice, Inbred C57BL , Mice, Knockout , Neurons/drug effects , Neurons/metabolism , Potassium Channels, Voltage-Gated/antagonists & inhibitors , TRPA1 Cation Channel/genetics , TRPA1 Cation Channel/metabolism , TRPM Cation Channels/genetics , TRPM Cation Channels/metabolism , TRPM Cation Channels/physiologyABSTRACT
Sirex nigricornis F. (Hymenoptera: Siricidae) is an innocuous pine-inhabiting woodwasp native to eastern North America, utilizing dead or dying pine trees as hosts. Although S. nigricornis F. does not cause economic damage, a closely related species, Sirex noctilio, was discovered in New York in 2004 and has continually spread throughout the northeastern United States and southern Canada, threatening the multi-billion-dollar pine timber industry of the southeastern United States and raising interest about potential interactions with native woodwasps and associated mortality agents. A non-sterilizing strain of the biological control agent, Deladenus siricidicola Bedding (Tylenchida: Neotylenchidae), was introduced along with S. noctilio but is not inhibiting the spread or establishment of S. noctilio A North American congener, Deladenus proximus Bedding, has been recently isolated from S. noctilio and shows promise as a biological control agent. To better understand the potential of D. proximus as a control agent for S. noctilio, we measured and dissected nearly 1,200 S. nigricornis females from Arkansas and Mississippi and evaluated differences among collection location with regard to nematode virulence, woodwasp body size, and egg load. Body size and egg load were related to collection location, and nematode infestation resulted in significantly smaller females who produced significantly fewer eggs. Female woodwasps, especially those collected in Arkansas, were often fully sterilized by nematodes, and a higher percent sterilization was inversely related to body size and fewer eggs. We propose field studies to test the nematode's ability to sterilize S. noctilio in the northeastern United States.
Subject(s)
Animal Distribution , Hymenoptera/parasitology , Tylenchida/physiology , Animals , Arkansas , Body Size , Female , Genetic Fitness , Geography , Hymenoptera/genetics , ReproductionABSTRACT
The hyo-laryngeal complex is a multi-segmented structure integrating the oral and pharyngeal cavities and thus a variety of critical functions related to airway control, feeding, and vocal communication. Currently, we lack a complete understanding of how the hyoid complex, and the functions it mediates, can also be affected by changes in surrounding cranio-facial dimensions. Here, we explore these relationships in a breed of domestic dog, the Portuguese Water Dog, which is characterized by strong cranio-facial variation. We used radiographic images of the upper body and head of 55 adult males and 51 adult females to obtain detailed measures of cranio-facial variation and hyoid anatomy. Principal components analysis revealed multiple orthogonal dimensions of cranio-facial variation, some of which were associated with significant differences in larynx position: the larynx occupied a more descended position in individuals with shorter, broader faces than in those with longer, narrower faces. We then tested the possibility that caudal displacement of the larynx in brachycephalic individuals might reflect a degree of tongue crowding resulting from facial shortening and reduction of oral and pharyngeal spaces. A cadaver sample was used to obtain detailed measurements of constituent bones of the hyoid skeleton and of the tongue body, and their relationships to cranio-facial size and shape and overall body size supported the tongue-crowding hypothesis. Considering the presence of descended larynges in numerous mammalian taxa, our findings establish an important precedent for the possibility that laryngeal descent can be initiated, and even sustained, in part in response to remodeling of the face and cranium for selective pressures unrelated to vocal production. These integrated changes could also have been involved in hominin evolution, where the different laryngeal positions in modern humans compared with nonhuman primates have been traditionally linked to the evolution of speech but which are likely to be multifactorial.
Subject(s)
Dogs/anatomy & histology , Larynx , Skull/anatomy & histology , Animals , Female , Larynx/diagnostic imaging , Male , Radiography , Skull/diagnostic imagingABSTRACT
A pressing need in neurobiology is the comprehensive identification and characterization of neuronal subclasses within the mammalian nervous system. To this end, we used constellation pharmacology as a method to interrogate the neuronal and glial subclasses of the mouse cerebellum individually and simultaneously. We then evaluated the data obtained from constellation-pharmacology experiments by cluster analysis to classify cells into neuronal and glial subclasses, based on their functional expression of glutamate, acetylcholine, and GABA receptors, among other ion channels. Conantokin peptides were used to identify N-methyl-d-aspartate (NMDA) receptor subtypes, which revealed that neurons of the young mouse cerebellum expressed NR2A and NR2B NMDA receptor subunits. Additional pharmacological tools disclosed differential expression of α-amino-3-hydroxy-5-methyl-4-isoxazloepropionic, nicotinic acetylcholine, and muscarinic acetylcholine receptors in different neuronal and glial subclasses. Certain cell subclasses correlated with known attributes of granule cells, and we combined constellation pharmacology with genetically labeled neurons to identify and characterize Purkinje cells. This study illustrates the utility of applying constellation pharmacology to classify neuronal and glial subclasses in specific anatomical regions of the brain.