RESUMEN
Environmental DNA (eDNA) has been established as a noninvasive and efficient approach to sample genetic material from aquatic environments. Although most commonly used to determine species presence and measure biodiversity, eDNA approaches also hold great potential to obtain population-level genetic information from water samples. In this study, we sequenced a panel of multiallelic microsatellite markers from filtered water and fish tissue samples to uncover patterns of intraspecific diversity in the freshwater Round Goby (Neogobius melanostomus) across their invaded range in the Laurentian Great Lakes region. Although we found that the concentration of nuclear eDNA is lower than mitochondrial eDNA, we nonetheless detected over two-thirds of all nuclear alleles identified from genotyped tissues in our eDNA samples, with the greatest recovery of common alleles in the population. Estimates of allele frequencies and genetic variability within and between populations were detected from eDNA in patterns that were consistent with individual tissue-based estimates of genetic diversity and differentiation. The strongest genetic differentiation in both eDNA and tissues exists in an isolation by distance pattern. Our study demonstrates the potential for eDNA-based approaches to characterize key population parameters required to effectively monitor, manage, or sustain aquatic species.
Asunto(s)
ADN Ambiental , Especies Introducidas , Animales , Lagos , Variación Genética , AguaRESUMEN
Advancements in environmental DNA (eDNA) approaches have allowed for rapid and efficient species detections in diverse environments. Although most eDNA research is focused on leveraging genetic diversity to identify taxa, some recent studies have explored the potential for these approaches to detect within-species genetic variation, allowing for population genetic assessments and abundance estimates from environmental samples. However, we currently lack a framework outlining the key considerations specific to generating, analysing and applying eDNA data for these two purposes. Here, we discuss how various genetic markers differ with regard to genetic information and detectability in environmental samples and how analysis of eDNA samples differs from common tissue-based analyses. We then outline how it may be possible to obtain species absolute abundance estimates from eDNA by detecting intraspecific genetic variation in mixtures of DNA under multiple scenarios. We also identify the major causes contributing to allele detection and frequency errors in eDNA data, discuss their consequences for population-level analyses and outline bioinformatic approaches to detect and remove erroneous sequences. This review summarizes the key advances required to harness the full potential of eDNA-based intraspecific genetic variation to inform population-level questions in ecology, evolutionary biology and conservation management.
Asunto(s)
ADN Ambiental , Biodiversidad , Código de Barras del ADN Taxonómico , Monitoreo del Ambiente , Genética de Población , Variación Genética/genéticaRESUMEN
The spread of nonindigenous species by shipping is a large and growing global problem that harms coastal ecosystems and economies and may blur coastal biogeographical patterns. This study coupled eukaryotic environmental DNA (eDNA) metabarcoding with dissimilarity regression to test the hypothesis that ship-borne species spread homogenizes port communities. We first collected and metabarcoded water samples from ports in Europe, Asia, Australia and the Americas. We then calculated community dissimilarities between port pairs and tested for effects of environmental dissimilarity, biogeographical region and four alternative measures of ship-borne species transport risk. We predicted that higher shipping between ports would decrease community dissimilarity, that the effect of shipping would be small compared to that of environment dissimilarity and shared biogeography, and that more complex shipping risk metrics (which account for ballast water and stepping-stone spread) would perform better. Consistent with our hypotheses, community dissimilarities increased significantly with environmental dissimilarity and, to a lesser extent, decreased with ship-borne species transport risks, particularly if the ports had similar environments and stepping-stone risks were considered. Unexpectedly, we found no clear effect of shared biogeography, and that risk metrics incorporating estimates of ballast discharge did not offer more explanatory power than simpler traffic-based risks. Overall, we found that shipping homogenizes eukaryotic communities between ports in predictable ways, which could inform improvements in invasive species policy and management. We demonstrated the usefulness of eDNA metabarcoding and dissimilarity regression for disentangling the drivers of large-scale biodiversity patterns. We conclude by outlining logistical considerations and recommendations for future studies using this approach.
Asunto(s)
ADN Ambiental , Ecosistema , ADN Ambiental/genética , Navíos , Biodiversidad , Agua , Monitoreo del Ambiente , Código de Barras del ADN TaxonómicoRESUMEN
A central focus of invasive species research has been on human efforts to eradicate invaders or reduce their abundance to mitigate the worst of their impacts. In some cases, however, populations of invasive species decline without human intervention, which may inform management responses to these invaders. Such is the case of the invasive rusty crayfish (Faxonius rusticus) in northern Wisconsin, USA, where systematic population monitoring since 1975 has revealed population declines in approximately half of the lakes surveyed. Population declines of invasive species without human intervention remain understudied, but there is even less research on how communities respond following such declines. Using 10 lakes in Vilas County, Wisconsin, we investigated community recovery of habitat (macrophytes) and prey (freshwater snails) of F. rusticus following up to 33 years of declines of this invader in some lakes using a dataset with a rare, long-term span over which consistent data were collected (1987, 2002, 2011, and 2020). We compared community responses in lakes where F. rusticus populations reached a peak and subsequently declined (boom-bust lakes) and lakes where our dataset only captured the decline of F. rusticus (bust lakes) to reference lakes with consistently high or low crayfish abundance over time. We found partial recovery of macrophytes and snails in the bust and boom-bust lakes where F. rusticus has declined, with recovery of macrophyte abundance and richness in the boom-bust lakes achieving levels observed in the low-crayfish reference lakes. Snail abundance and richness increased after declines of F. rusticus, though not to the level of the low-crayfish reference lakes, suggesting that snail recovery may lag macrophyte recovery because snails are dependent on macrophytes and associated periphyton for habitat. The recovery we document potentially represents long-term ecosystem resilience of lakes to biological invasions. Our results suggest that lake communities may recover without active restoration interventions after invasive crayfish population declines, although identifying which lakes experience these natural declines remains a priority for future research and management.
Asunto(s)
Astacoidea , Ecosistema , Humanos , Animales , Astacoidea/fisiología , Lagos , Alimentos Marinos , Caracoles , Especies IntroducidasRESUMEN
Advances in environmental DNA (eDNA) methodologies have led to improvements in the ability to detect species and communities in aquatic environments, yet the majority of studies emphasize biological diversity at the species level by targeting variable sites within the mitochondrial genome. Here, we demonstrate that eDNA approaches also have the capacity to detect intraspecific diversity in the nuclear genome, allowing for assessments of population-level allele frequencies and estimates of the number of genetic contributors in an eDNA sample. Using a panel of microsatellite loci developed for the round goby (Neogobius melanostomus), we tested the similarity between eDNA-based and individual tissue-based estimates of allele frequencies from experimental mesocosms and in a field-based trial. Subsequently, we used a likelihood-based DNA mixture framework to estimate the number of unique genetic contributors in eDNA samples and in simulated mixtures of alleles. In both mesocosm and field samples, allele frequencies from eDNA were highly correlated with allele frequencies from genotyped round goby tissue samples, indicating nuclear markers can be reliably amplified from water samples. DNA mixture analyses were able to estimate the number of genetic contributors from mesocosm eDNA samples and simulated mixtures of DNA from up to 58 individuals, with the degree of positive or negative bias dependent on the filtering scheme of low-frequency alleles. With this study we document the application of eDNA and multiple amplicon-based methods to obtain intraspecific nuclear genetic information and estimate the absolute abundance of a species in eDNA samples. With proper validation, this approach has the potential to advance noninvasive survey methods to characterize populations and detect population-level genetic diversity.
Asunto(s)
ADN Ambiental , Peces , Animales , Biodiversidad , Peces/genética , Frecuencia de los Genes , Humanos , Funciones de VerosimilitudRESUMEN
Effective engagement among scientists, government agency staff, and policymakers is necessary for solving fisheries challenges, but remains challenging for a variety of reasons. We present seven practices learned from a collaborative project focused on invasive species in the Great Lakes region (USA-CAN). These practices were based on a researcher-manager model composed of a research team, a management advisory board, and a bridging organization. We suggest this type of system functions well when (1) the management advisory board is provided compelling rationale for engagement; (2) the process uses key individuals as communicators; (3) the research team thoughtfully selects organizations and individuals involved; (4) the funding entity provides logistical support and allows for (5) a flexible structure that prioritizes management needs; (6) a bridging organization sustains communication between in-person meetings; and (7) the project team determines and enacts a project endpoint. We predict these approaches apply equally effectively to other challenges at the research-management-policy interface, including reductions of water pollution, transitions to renewable energy, increasing food security, and addressing climate change.
RESUMEN
The genomic revolution has fundamentally changed how we survey biodiversity on earth. High-throughput sequencing ("HTS") platforms now enable the rapid sequencing of DNA from diverse kinds of environmental samples (termed "environmental DNA" or "eDNA"). Coupling HTS with our ability to associate sequences from eDNA with a taxonomic name is called "eDNA metabarcoding" and offers a powerful molecular tool capable of noninvasively surveying species richness from many ecosystems. Here, we review the use of eDNA metabarcoding for surveying animal and plant richness, and the challenges in using eDNA approaches to estimate relative abundance. We highlight eDNA applications in freshwater, marine and terrestrial environments, and in this broad context, we distill what is known about the ability of different eDNA sample types to approximate richness in space and across time. We provide guiding questions for study design and discuss the eDNA metabarcoding workflow with a focus on primers and library preparation methods. We additionally discuss important criteria for consideration of bioinformatic filtering of data sets, with recommendations for increasing transparency. Finally, looking to the future, we discuss emerging applications of eDNA metabarcoding in ecology, conservation, invasion biology, biomonitoring, and how eDNA metabarcoding can empower citizen science and biodiversity education.
Asunto(s)
Biodiversidad , Código de Barras del ADN Taxonómico/métodos , Ecología/métodos , Animales , Conservación de los Recursos Naturales , Cartilla de ADN , Monitoreo del Ambiente , PlantasRESUMEN
Parasites can alter communities by reducing densities of keystone hosts, but few studies have examined how trait-mediated indirect effects of parasites can alter ecological communities. We test how trematode parasites (Microphallus spp.) that affect invasive crayfish (Orconectes rusticus) behavior alter how crayfish impact lake littoral communities. O. rusticus drive community composition in north temperate lakes, and predatory fish can reduce crayfish activity and feeding. In laboratory studies, Microphallus parasites also alter O. rusticus behavior: infected O. rusticus eat fewer macroinvertebrates and are bolder near predatory fish than uninfected individuals. We used a 2 x 2 factorial experiment to test how predatory fish and parasites affect O. rusticus impacts in large mesocosms over 4 weeks. We predicted (1) that when predators were absent, infected crayfish would have lower impacts than uninfected crayfish on macrophytes and macroinvertebrates (as well as reduced growth and higher mortality). However, (2) when predators were present but unable to consume crayfish, infected crayfish would have greater impacts (as well as greater growth and lower mortality) than uninfected crayfish because of increased boldness. Because of its effect on crayfish feeding behavior, we also predicted (3) that infection would alter macrophyte and macroinvertebrate community composition. In contrast to our first hypothesis, we found that infected and uninfected crayfish had similar impacts on lower trophic levels when predators were absent. Across all treatments, infected crayfish were more likely to be outside shelters and had greater growth than uninfected crayfish, suggesting that the reduced feeding observed in short-term experiments does not occur over longer timescales. However, in support of the second hypothesis, when predatory fish were present, infected crayfish ate more macroinvertebrates than did uninfected crayfish, likely due to increased boldness. We also observed a trend for greater macrophyte consumption associated with infection and a trend indicating infection might alter macroinvertebrate community composition. Our results suggest that parasites can alter aquatic communities in mesocosms merely by modifying host behavior.
Asunto(s)
Astacoidea/parasitología , Ecosistema , Agua Dulce , Especies Introducidas , Trematodos/fisiología , Animales , Conducta Animal , Interacciones Huésped-ParásitosRESUMEN
The roles of both kainate receptors (KARs) and metabotropic glutamate receptors (mGluRs) in mossy fiber long-term potentiation (MF-LTP) have been extensively studied in hippocampal brain slices, but the findings are controversial. In this study, we have addressed the roles of both mGluRs and KARs in MF-LTP in anesthetized rats. We found that MF-LTP could be induced in the presence of either GluK1-selective KAR antagonists or group I mGluR antagonists. However, LTP was inhibited when the group I mGluRs and the GluK1-KARs were simultaneously inhibited. Either mGlu1 or mGlu5 receptor activation is sufficient to induce this form of LTP as selective inhibition of either subtype alone, together with the inhibition of KARs, did not inhibit MF-LTP. These data suggest that mGlu1 receptors, mGlu5 receptors, and GluK1-KARs are all engaged during high-frequency stimulation, and that the activation of any one of these receptors alone is sufficient for the induction of MF-LTP in vivo.
Asunto(s)
Potenciación a Largo Plazo/fisiología , Fibras Musgosas del Hipocampo/fisiología , Receptor del Glutamato Metabotropico 5/fisiología , Receptores de Ácido Kaínico/fisiología , Receptores de Glutamato Metabotrópico/fisiología , Animales , Potenciación a Largo Plazo/efectos de los fármacos , Masculino , Fibras Musgosas del Hipocampo/efectos de los fármacos , Ratas , Ratas Wistar , Receptor del Glutamato Metabotropico 5/antagonistas & inhibidores , Receptores de Ácido Kaínico/antagonistas & inhibidores , Receptores de Glutamato Metabotrópico/antagonistas & inhibidoresRESUMEN
Identifying which nonindigenous species will become invasive and forecasting the damage they will cause is difficult and presents a significant problem for natural resource management. Often, the data or resources necessary for ecological risk assessment are incomplete or absent, leaving environmental decision makers ill equipped to effectively manage valuable natural resources. Structured expert judgment (SEJ) is a mathematical and performance-based method of eliciting, weighting, and aggregating expert judgments. In contrast to other methods of eliciting and aggregating expert judgments (where, for example, equal weights may be assigned to experts), SEJ weights each expert on the basis of his or her statistical accuracy and informativeness through performance measurement on a set of calibration variables. We used SEJ to forecast impacts of nonindigenous Asian carp (Hypophthalmichthys spp.) in Lake Erie, where it is believed not to be established. Experts quantified Asian carp biomass, production, and consumption and their impact on 4 fish species if Asian carp were to become established. According to experts, in Lake Erie Asian carp have the potential to achieve biomass levels that are similar to the sum of biomasses for several fishes that are harvested commercially or recreationally. However, the impact of Asian carp on the biomass of these fishes was estimated by experts to be small, relative to long term average biomasses, with little uncertainty. Impacts of Asian carp in tributaries and on recreational activities, water quality, or other species were not addressed. SEJ can be used to quantify key uncertainties of invasion biology and also provide a decision-support tool when the necessary information for natural resource management and policy is not available.
Asunto(s)
Carpas/fisiología , Conservación de los Recursos Naturales/métodos , Especies Introducidas , Animales , Explotaciones Pesqueras , Peces/fisiología , Predicción , Humanos , Lagos , Ontario , Dinámica Poblacional , Recreación , Medición de Riesgo , Estados UnidosRESUMEN
Invasive species introduced via the ballast water of commercial ships cause enormous environmental and economic damage worldwide. Accurate monitoring for these often microscopic and morphologically indistinguishable species is challenging but critical for mitigating damages. We apply eDNA sampling, which involves the filtering and subsequent DNA extraction of microscopic bits of tissue suspended in water, to ballast and harbor water sampled during a commercial ship's 1400 km voyage through the North American Great Lakes. Using a lab-based gel electrophoresis assay and a rapid, field-ready light transmission spectroscopy (LTS) assay, we test for the presence of two invasive species: quagga (Dreissena bugensis) and zebra (D. polymorpha) mussels. Furthermore, we spiked a set of uninfested ballast and harbor samples with zebra mussel tissue to further test each assay's detection capabilities. In unmanipulated samples, zebra mussel was not detected, while quagga mussel was detected in all samples at a rate of 85% for the gel assay and 100% for the LTS assay. In the spiked experimental samples, both assays detected zebra mussel in 94% of spiked samples and 0% of negative controls. Overall, these results demonstrate that eDNA sampling is effective for monitoring ballast-mediated invasions and that LTS has the potential for rapid, field-based detection.
Asunto(s)
ADN/análisis , Dreissena , Especies Introducidas , Navíos , Agua/análisis , Animales , Bivalvos/genética , Reacción en Cadena de la Polimerasa , Análisis EspectralRESUMEN
Glutamate, the primary excitatory neurotransmitter in the central nervous system (CNS), exerts neuromodulatory actions via the activation of metabotropic glutamate (mGlu) receptors. There are eight known mGlu receptor subtypes (mGlu1-8), which are widely expressed throughout the brain, and are divided into three groups (I-III), based on signalling pathways and pharmacological profiles. Group III mGlu receptors (mGlu4/6/7/8) are primarily, although not exclusively, localised on presynaptic terminals, where they act as both auto- and hetero-receptors, inhibiting the release of neurotransmitter. Until recently, our understanding of the role of individual group III mGlu receptor subtypes was hindered by a lack of subtype-selective pharmacological tools. Recent advances in the development of both orthosteric and allosteric group III-targeting compounds, however, have prompted detailed investigations into the possible functional role of these receptors within the CNS, and revealed their involvement in a number of pathological conditions, such as epilepsy, anxiety and Parkinson's disease. The heterogeneous expression of group III mGlu receptor subtypes throughout the brain, as well as their distinct distribution at glutamatergic and GABAergic synapses, makes them ideal targets for therapeutic intervention. This review summarises the advances in subtype-selective pharmacology, and discusses the individual roles of group III mGlu receptors in physiology, and their potential involvement in disease.
Asunto(s)
Receptores de Glutamato Metabotrópico/efectos de los fármacos , Sitio Alostérico , Hipocampo/fisiología , Humanos , Ligandos , Enfermedades del Sistema Nervioso/fisiopatología , Plasticidad Neuronal/fisiología , Conformación Proteica , Receptores de Glutamato Metabotrópico/química , Receptores de Glutamato Metabotrópico/fisiología , Transmisión Sináptica/fisiologíaRESUMEN
Recently, authors have theorized that invasive species prevention is more cost-effective than control in protecting ecosystem services. However, quantification of the effectiveness of prevention is rare because experiments at field scales are expensive or infeasible. We therefore used structured expert judgment to quantify the efficacy of 17 proposed strategies to prevent Asian carp invasion of the Laurentian Great Lakes via the hydrologic connection between the Mississippi and Great Lakes watersheds. Performance-weighted expert estimates indicated that hydrologic separation would prevent 99% (95,100; median, 5th and 95th percentiles) of Asian carp access, while electric and acoustic-bubble-strobe barriers would prevent 92% (85,95) and 92% (75,95), respectively. For all other strategies, estimated effectiveness was lower, with greater uncertainty. When potential invasions by other taxa are considered, the effectiveness of hydrologic separation increases relative to strategies that are effective primarily for fishes. These results could help guide invasive species management in many waterways globally.
Asunto(s)
Carpas/fisiología , Hidrología , Especies Introducidas , Juicio , Lagos , Animales , Calibración , Geografía , Mississippi , RíosRESUMEN
Environmental DNA (eDNA) surveillance holds great promise for improving species conservation and management. However, few studies have investigated eDNA dynamics under natural conditions, and interpretations of eDNA surveillance results are clouded by uncertainties about eDNA degradation. We conducted a literature review to assess current understanding of eDNA degradation in aquatic systems and an experiment exploring how environmental conditions can influence eDNA degradation. Previous studies have reported macrobial eDNA persistence ranging from less than 1 day to over 2 weeks, with no attempts to quantify factors affecting degradation. Using a SYBR Green quantitative PCR assay to observe Common Carp ( Cyprinus carpio ) eDNA degradation in laboratory mesocosms, our rate of Common Carp eDNA detection decreased over time. Common Carp eDNA concentration followed a pattern of exponential decay, and observed decay rates exceeded previously published values for aquatic macrobial eDNA. Contrary to our expectations, eDNA degradation rate declined as biochemical oxygen demand, chlorophyll, and total eDNA (i.e., from any organism) concentration increased. Our results help explain the widely divergent, previously published estimates for eDNA degradation. Measurements of local environmental conditions, consideration of environmental influence on eDNA detection, and quantification of local eDNA degradation rates will help interpret future eDNA surveillance results.
Asunto(s)
ADN/química , Agua Dulce/química , Animales , Carpas , Ambiente , Reacción en Cadena de la PolimerasaRESUMEN
Nonindigenous species can cause major changes to community interactions and ecosystem processes. The strong impacts of these species are often attributed to their high demographic success. While the importance of enemy release in facilitating invasions has often been emphasized, few studies have addressed the role of parasites in the invasive range in controlling demographic success of potential invaders. Here we examine whether a trematode parasite (Microphallus spp.) can contribute to previously documented alternate states in the abundance of invasive rusty crayfish (Orconectes rusticus) in north temperate lakes in Wisconsin, USA. Microphallus infect O. rusticus after emerging from their first intermediate host, a hydrobiid snail. As previously documented, O. rusticus reduce densities of hydrobiid snails through direct predation and destruction of macrophyte habitat. Therefore, if Microphallus substantially reduce O. rusticus fitness, these parasites may reinforce a state of low crayfish abundance, and, at the other extreme, abundant crayfish may repress these parasites, reinforcing a state of high crayfish abundance. From samples collected from 109 sites in 16 lakes, we discovered (1) a positive relationship between crayfish infection intensity and hydrobiid snail abundance, (2) a negative relationship between parasite prevalence and crayfish abundance, and (3) a negative relationship between parasite prevalence and crayfish population growth. With experiments, we found that infection with Microphallus reduced foraging behavior and growth in O. rusticus, which may be the mechanisms responsible for the population reductions we observed. Overall results are consistent with the hypothesis that Microphallus contributes to alternate states in the abundance and impacts of O. rusticus.
Asunto(s)
Astacoidea/parasitología , Conducta Alimentaria , Especies Introducidas , Trematodos/fisiología , Animales , Astacoidea/crecimiento & desarrollo , Astacoidea/fisiología , Ecosistema , Lagos , Dinámica Poblacional , Caracoles/parasitología , WisconsinRESUMEN
Many ecosystems appear subject to regime shifts--abrupt changes from one state to another after crossing a threshold or tipping point. Thresholds and their associated stability landscapes are determined within a coupled socioeconomic-ecological system (SES) where human choices, including those of managers, are feedback responses. Prior work has made one of two assumptions about managers: that they face no institutional constraints, in which case the SES may be managed to be fairly robust to shocks and tipping points are of little importance, or that managers are rigidly constrained with no flexibility to adapt, in which case the inferred thresholds may poorly reflect actual managerial flexibility. We model a multidimensional SES to investigate how alternative institutions affect SES stability landscapes and alter tipping points. With institutionally dependent human feedbacks, the stability landscape depends on institutional arrangements. Strong institutions that account for feedback responses create the possibility for desirable states of the world and can cause undesirable states to cease to exist. Intermediate institutions interact with ecological relationships to determine the existence and nature of tipping points. Finally, weak institutions can eliminate tipping points so that only undesirable states of the world remain.
Asunto(s)
Conservación de los Recursos Naturales/métodos , Ecología/métodos , Ecosistema , Ambiente , Modelos Teóricos , Política Pública , Animales , Astacoidea/fisiología , Lubina/fisiología , Explotaciones Pesqueras/métodos , Humanos , Factores SocioeconómicosRESUMEN
Since Ficetola et al. (2008) alerted ecologists and conservation biologists to the existence of environmental DNA (eDNA), the number of studies using eDNA has exploded, with a rapidly increasing diversity of research, monitoring, and management objectives. Initial applications focused on amphibians and fishes while today's taxonomic targets span the phylogenetic tree. The environmental media that are sampled have expanded from freshwater to saltwater to soils, and, most recently, to air. In this issue of Molecular Ecology Resources, Lynggaard et al. (Molecular Ecology Resources, 2023) use eDNA captured on air filters to census vertebrate biodiversity in a forest. With a three day, six sample period, 143 sample effort in a nature park in a rural area of Zealand, Denmark, their wild species detections comprised about 25% of the terrestrial vertebrates that are known to occur in the area, including about 33% of the mammal, 17% of the bird, and 60% of the amphibian species. This study demonstrates that air sampling for eDNA has the potential to become a powerful standard method for terrestrial biodiversity assessment that is complementary to traditional methods (e.g., trapping, visual and acoustic observation, collection of scat and hair).
Asunto(s)
ADN Ambiental , Monitoreo del Ambiente , Animales , Filogenia , Monitoreo del Ambiente/métodos , Código de Barras del ADN Taxonómico/métodos , Vertebrados/genética , Biodiversidad , Peces/genética , Anfibios/genética , Mamíferos/genética , EcosistemaRESUMEN
Traditionally, neurotransmitters are associated with a fast, or phasic, type of action on neurons in the central nervous system (CNS). However, accumulating evidence indicates that γ-aminobutyric acid (GABA) and glutamate can also have a continual, or tonic, influence on these cells. Here, in voltage- and current-clamp recordings in rat brain slices, we identify three types of tonically active receptors in a single CNS structure, the thalamic reticular nucleus (TRN). Thus, TRN contains constitutively active GABAA receptors (GABAA Rs), which are located on TRN neurons and generate a persistent outward Cl(-) current. When TRN neurons are depolarized, blockade of this current increases their action potential output in response to current injection. Furthermore, TRN contains tonically active GluN2B-containing N-methyl-D-aspartate receptors (NMDARs). These are located on reticuloreticular GABAergic terminals in TRN and generate a persistent facilitation of vesicular GABA release from these terminals. In addition, TRN contains tonically active metabotropic glutamate type 2 receptors (mGlu2Rs). These are located on glutamatergic cortical terminals in TRN and generate a persistent reduction of vesicular glutamate release from these terminals. Although tonically active GABAA Rs, NMDARs and mGlu2Rs operate through different mechanisms, we propose that the continual and combined activity of these three receptor types ultimately serves to hyperpolarize TRN neurons, which will differentially affect the output of these cells depending upon the current state of their membrane potential. Thus, when TRN cells are relatively depolarized, their firing in single-spike tonic mode will be reduced, whereas when these cells are relatively hyperpolarized, their ability to fire in multispike burst mode will be facilitated.
Asunto(s)
Potenciales de Acción , Núcleos Talámicos Intralaminares/fisiología , Receptores de GABA-A/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Animales , Cloruros/metabolismo , Ácido Glutámico/metabolismo , Núcleos Talámicos Intralaminares/metabolismo , Neuronas/metabolismo , Neuronas/fisiología , Terminales Presinápticos/metabolismo , Terminales Presinápticos/fisiología , Ratas , Ratas Wistar , Vías Secretoras , Vesículas Sinápticas/metabolismo , Vesículas Sinápticas/fisiología , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Group II metabotropic receptors (mGluRs) regulate central synaptic transmission by modulating neurotransmitter release. However, the lack of pharmacological tools differentiating between mGlu2 and mGlu3 receptors has hampered identification of the roles of these two receptor subtypes. We have used LY395756 [(1SR,2SR,4RS,5RS,6SR)-2-amino-4-methylbicyclo[3.1.0]-hexane2,6-dicarboxylic], an agonist at mGlu2 receptors and an antagonist at mGlu3 receptors in cell lines, to investigate the roles of these receptors in the temporo-ammonic path from entorhinal cortex to CA1-stratum lacunosum moleculare in rat hippocampal slices. Surprisingly, the degree of inhibition of the field EPSP induced by LY395756 fell into two distinct groups, with EC(50) values of <1 µm and >100 µm. In "sensitive" slices, LY395756 had additive actions with a mixed mGlu2/mGlu3 agonist, DCG-IV [(2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine], whereas in "insensitive" slices, LY395756 reduced the effect of DCG-IV, with an IC(50) of â¼1 µm. This separation into sensitive and insensitive slices could be explained by LY395756 acting as an mGlu2 agonist and mGlu3 antagonist, respectively, a finding supported by data from mice lacking these receptors. The heterogeneity was correlated with differences in expression levels of mGlu2 receptors within our Wistar colony and other Wistar substrains. The initial search for a behavioral correlate indicated that rats lacking mGlu2 receptors showed anxiety-like behavior in open-field and elevated plus maze assays. These findings have implications for rat models of psychiatric disease and are especially pertinent given that mGlu2 receptors are targets for compounds under development for anxiety.