Inhibitory synaptic mechanisms oppose epileptic network activity in the brain. The breakdown in this inhibitory restraint and propagation of seizure activity has been linked to the overwhelming of ...feedforward inhibition, which is provided in large part by parvalbumin-expressing (PV) interneurons in the cortex. The underlying cellular processes therefore represent potential targets for understanding and preventing the propagation of seizure activity. Here we use an optogenetic strategy to test the hypothesis that depolarization block in PV interneurons is a significant factor during the loss of inhibitory restraint. Depolarization block results from the inactivation of voltage-gated sodium channels and leads to impaired action potential firing. We used focal NMDA stimulation to elicit reproducible epileptiform discharges in hippocampal organotypic brain slices from male and female mice and combined this with targeted recordings from defined neuronal populations. Simultaneous patch-clamp recordings from PV interneurons and pyramidal neurons revealed epileptiform activity that was associated with an overwhelming of inhibitory synaptic mechanisms and the emergence of a partial, and then complete, depolarization block in PV interneurons. To counteract this depolarization block, we developed protocols for eliciting pulsed membrane hyperpolarization via the inhibitory opsin, archaerhodopsin. This optical approach was effective in counteracting cumulative inactivation of voltage-gated channels, maintaining PV interneuron action potential firing properties during the inhibitory restraint period, and reducing the probability of initiating epileptiform activity. These experiments support the idea that depolarization block is a point of weakness in feedforward inhibitory synaptic mechanisms and represents a target for preventing the initiation and spread of seizure activity.
GABA
receptor-mediated synaptic transmission opposes seizure activity by establishing an inhibitory restraint against spreading excitation. Parvalbumin-expressing (PV) interneurons contribute significantly to this inhibitory restraint, but it has been suggested that these cells are overwhelmed as they enter a state of "depolarization block." Here we test the importance of this process by devising an optogenetic strategy to selectively relieve depolarization block in PV interneurons. By inducing brief membrane hyperpolarization, we show that it is possible to reduce depolarization block in PV interneurons, maintain their action potential firing in the face of strong excitation, and disrupt epileptiform activity in an
model. This represents a proof of principle that targeting rate-limiting processes can strengthen the inhibitory restraint of epileptiform activity.
Romanian flora includes a high diversity of orchids, any with a high conservationist interest, but many 0f themnot yet sufficiently investigated from an ecological, phytosociological or genetic point ...of view. Phenotypically analysis is also scarce and only a few data have been published in relation to the effects of site factors on the morphological and anatomical characteristics of the family Orchidaceae. This study analyses the variability of two populations of Anacamptis coriophora, based on their coenological affiliations, morphological observations and biometrical measurements. The data were collected from two grassland areas in Romania Stoenesti, Vâlcea County and Cobia, Dâmbovița County. The results showed that both populations are influenced by temperature, light and humidity and develop normally under optimal conditions. In the most favorable conditions encountered in Vâlcea county, orchids offer the splendor of flowers for a long time. Orchids in our country usually bloom in March, sometimes lasting until August, they have a delicate appearance, varied colors and the most diverse perfumes. The information obtained regarding the morphological diversity of the two populations could be suitable for designing strategies for their conservation in both locations.
Acute hepatopancreatic necrosis disease (AHPND), caused by marine bacteria Vibrio Parahaemolyticus, is a huge problem in shrimp farms. The V. parahaemolyticus infecting material is contained in a ...plasmid which encodes for the lethal toxins PirABVp, whose primary target tissue is the hepatopancreas, causing sloughing of epithelial cells, necrosis, and massive hemocyte infiltration. To get a better understanding of the hepatopancreas response during AHPND, juvenile shrimp Litopenaeus vannamei were infected by immersion with V. parahaemolyticus. We performed transcriptomic mRNA sequencing of infected shrimp hepatopancreas, at 24 hours post-infection, to identify novel differentially expressed genes a total of 174,098 transcripts were examined of which 915 transcripts were found differentially expressed after comparative transcriptomic analysis: 442 up-regulated and 473 down-regulated transcripts. Gene Ontology term enrichment analysis for up-regulated transcripts includes metabolic process, regulation of programmed cell death, carbohydrate metabolic process, and biological adhesion, whereas for down-regulated transcripts include, microtubule-based process, cell activation, and chitin metabolic process. The analysis of protein- protein network between up and down-regulated genes indicates that the first gene interactions are connected to oxidation-processes and sarcomere organization. Additionally, protein-protein networks analysis identified 20-top highly connected hub nodes. Based on their immunological or metabolic function, ten candidate transcripts were selected to measure their mRNA relative expression levels in AHPND infected shrimp hepatopancreas by RT-qPCR. Our results indicate a close connection between the immune and metabolism systems during AHPND infection. Our RNA-Seq and RT-qPCR data provide the possible immunological and physiological scenario as well as the molecular pathways that take place in the shrimp hepatopancreas in response to an infectious disease.
The immunostimulatory potential of the marine yeast Yarrowia lipolytica (D1 and N6 strains) administered orally was evaluated in the white shrimp Litopenaeus vannamei. Yeasts and commercial glucans ...were mixed with a commercial feed to formulate diets with a 1.1% concentration of immunostimulants. The shrimp were fed daily for a period of 21 days. Weekly determinations were performed for immunological parameters in hemolymph, such as total hemocyte count (THC), lysozyme activity (LYZ), prophenoloxidase activity, antioxidant enzymatic activities (superoxide dismutase SOD, catalase CAT, and peroxidases), and bactericidal activity against Vibrio parahaemolyticus. Expression profiles of penaeidin (PEN), lysozyme (LYZ), and prophenoloxidase (proPO) immune genes were evaluated in hemocytes. In general, an increase in the immune parameters was observed in shrimp fed yeast diet compared to glucan and the control diets. Yarrowia lipolytica, especially strain N6, provided maximum immunostimulatory effects evidenced by the increase of immune parameters (THC, LYZ, SOD, CAT) and gene expression profile. In conclusion, this study demonstrated that Y. lipolytica had immunostimulatory effects and increased bactericidal activity in L. vannamei hemocytes against V. parahaemolyticus. These findings open the path for the potential application of Y. lipolytica-based immunostimulant for shrimp aquaculture.
•This is the first report on the immunological response to Yarrowia lipolytica in shrimp.•Oral administration of Y. lipolytica stimulates innate immune responses.•Y. lipolytica increased bactericidal activity in shrimp.•Circulating hemocytes and immune-related enzymatic activities increased in shrimp fed Y. lipolytica.•Immune-related gene expression up-regulated in hemocytes upon yeast administration.
This paper presents an original and effective application of support vector clustering (SVC) to electrical load pattern classification. The proposed SVC-based approach combines the calculation of the ...support vectors, carried out by using a classical procedure adopting a Gaussian kernel, with a specifically developed deterministic algorithm to form the clusters. This algorithm exploits the meaningful location of the bounded support vectors (BSVs) to define the outliers, identifying the clusters in function of the distance of the non-BSVs to the BSVs. Its implementation is less computationally intensive than other existing approaches and the cluster formation is driven by a single user-defined threshold. Extended comparison to other clustering methods is included to show the effectiveness of the proposed approach in grouping multidimensional load pattern data into non-overlapping clusters. This effectiveness is confirmed by the calculation of various cluster validity indicators. In particular, the most successful tasks are the identification of the outliers and the more effective formation of small numbers of clusters with respect to other methods.
Anatomically plausible networks of functionally inter-connected regions have been reliably demonstrated at rest, although the neurochemical basis of these 'resting state networks' is not well ...understood. In this study, we combined magnetic resonance spectroscopy (MRS) and resting state fMRI and demonstrated an inverse relationship between levels of the inhibitory neurotransmitter GABA within the primary motor cortex (M1) and the strength of functional connectivity across the resting motor network. This relationship was both neurochemically and anatomically specific. We then went on to show that anodal transcranial direct current stimulation (tDCS), an intervention previously shown to decrease GABA levels within M1, increased resting motor network connectivity. We therefore suggest that network-level functional connectivity within the motor system is related to the degree of inhibition in M1, a major node within the motor network, a finding in line with converging evidence from both simulation and empirical studies. DOI: http://dx.doi.org/10.7554/eLife.01465.001.
The escape response in some decapod crustaceans through tail-flipping represents an excellent model of short-term, exhausting muscular activity to understand the role of energy depletion and ...oxidative stress in muscle fatigue. In this study indicators of oxidative stress and changes in energy metabolism were measured during escape response until fatigue in whiteleg shrimp Litopenaeus vannamei. The escape response of individual shrimp was elicited in a tank by prodding the shrimp with a stick until exhaustion (no further response). A control group consisted of undisturbed shrimp, and a third group was allowed to recover. Arginine phosphate (ArgP) and adenylic energy charge (AEC) were measured by reverse phase HPLC, while fuels, reserves, antioxidant enzyme activities (superoxide dismutase, catalase, glutathione peroxidase-GPx), thiobarbituric acid reactive substances, and protein carbonyl content were analyzed by spectrophotometric methods. Shrimp reached fatigue after 31 tail-flips, which decreased abdominal muscle ArgP by 75% and AEC by 18%. Hemolymph lactate levels increased 5 times and muscle GPx activity by 2.7 times. After 1h of recovery, ArgP and AEC base levels were re-established, while hemolymph lactate concentration and GPx activity were still increased, and glucose hemolymph content increased 1.8 times. The drastic decrease in ArgP levels prevents a drop of AEC to a critical level, in accordance with a situation of non-lethal stress. Although no significant changes were observed in muscle glycogen or lactate concentrations, the increase in hemolymph lactate levels indicates an important contribution of muscle glycolysis to prevent a more pronounced decrease in ATP and AEC. The increase in GPx activity in abdominal muscle suggests an increase in reactive oxygen species production during intensive muscular activity, which was adequately neutralized, for no signs of oxidative damage occurred, as indicated by the lack of lipid and protein oxidation. These responses suggest adequate energy efficiency and prevention of oxidative damage during the escape response, in accordance with the adaptative value of such behavior to avoid instantaneous danger, with a capacity of rapid recovery.
•Escape response Bioenergetics and oxidative stress of L. vannamei were studied.•Escape response to exhaustion decreased energetic status and phosphoarginine levels.•Escape response to exhaustion increased hemolymph lactate.•After recovery the energetic status was reestablished and glucose hemolymph increased.•Glutathione peroxidase activity increased after exhaustion and after 1h recovery.
Display omitted
•Three stages of E-RLO infection were identified based on internal lesions.•The intensity of infection differed between tissues.•Damage suggests that the E-RLO has the capacity to ...impairs the C. gigas.
Rickettsia infection in adult Japanese oysters (Crassostrea gigas) was observed in 2015 at San Ignacio Lagoon, Baja California Sur, Mexico and characterized using molecular tools. In the present study, the degree of infection by Eosinophilic Rickettsia-Like Organism (E-RLO), characterized by intracellular inclusions in gill, mantle, labial palps, digestive tract and gonadal ducts, was evaluated using histological methods and was associated with visible injuries on body surface, such as blisters, shell damage and necrosis. Most of the oysters (92.2%) had some type of observable symptoms; 90.2% of all oysters had blisters in the mantle, and all oysters presented with E-RLO in the epithelia of at least one of the analyzed tissues (gills, labial palps, mantle, gonadal ducts and digestive tract). The highest intensity of infection (percent of E-RLO coverage) was observed in the labial palps (18.6%) and the lowest in digestive tract (2.6% of the examined tissue). Oysters with external injuries had higher infection intensities than the oysters without external injuries. Considering the clinical signs (observable symptoms and histological findings), we propose three infective stages of E-RLO disease development, a scale that could be used for early detection.
Biomarkers for monitoring shrimp health have been proposed but scarcely evaluated at the farm level. We analyzed several indicators of energy status in shrimp under farming conditions concerning ...stocking densities (100 m-2 with biofloc, 15 and 7 m-2). The influence of the year's season (temperature) was also analyzed, and, finally, an unfortunate event of White spot syndrome virus (WSSV) infection occurred on the 7 m-2 farms, adding another condition. At shrimp sampling from ponds, the effect of acute handling stress was also analyzed for indicators typically affected by such procedure with a 3- and 10-fold increase in glucose and lactate levels in hemolymph, respectively, regardless of density. This response was partially blunted at lower temperatures and WSSV incidence. Increased levels of protein in the hepatopancreas, adenylic energy charge (AEC) in both hepatopancreas and muscle and phosphagens in muscle were observed in shrimp from the 100 m-2 farms, suggesting a better nutritional and energetic status in shrimp cultured at high density with biofloc technology. Shrimp with WSSV presented lower hemocyanin levels in hemolymph, most likely associated with its role in the immune response. In WSSV-infected shrimp, the stress response regarding glucose increase was blunted, whereas a stress-induced increase in triglycerides (TG) levels in hemolymph was observed only with WSSV. Increased TG levels in those shrimp hepatopancreas could indicate a switch from carbohydrate to lipid-based metabolism associated with the preferential use of carbohydrates (Warburg effect) for virus replication in the early infection state.
Current anti-epileptic medications that boost synaptic inhibition are effective in reducing several types of epileptic seizure activity. Nevertheless, these drugs can generate significant ...side-effects and even paradoxical responses due to the broad nature of their action. Recently developed chemogenetic techniques provide the opportunity to pharmacologically recruit endogenous inhibitory mechanisms in a selective and circuit-specific manner. Here, we use chemogenetics to assess the potential of suppressing epileptiform activity by enhancing the synaptic output from three major interneuron populations in the rodent hippocampus: parvalbumin (PV), somatostatin (SST), and vasoactive intestinal peptide (VIP) expressing interneurons. To target different neuronal populations, promoter-specific cre-recombinase mice were combined with viral-mediated delivery of chemogenetic constructs. Targeted electrophysiological recordings were then conducted in an
model of chronic, drug-resistant epilepsy. In addition, behavioral video-scoring was performed in an
model of acutely triggered seizure activity. Pre-synaptic and post-synaptic whole cell recordings in brain slices revealed that each of the three interneuron types increase their firing rate and synaptic output following chemogenetic activation. However, the interneuron populations exhibited different effects on epileptiform discharges. Recruiting VIP interneurons did not change the total duration of epileptiform discharges. In contrast, recruiting SST or PV interneurons produced robust suppression of epileptiform synchronization. PV interneurons exhibited the strongest effect per cell, eliciting at least a fivefold greater reduction in epileptiform activity than the other cell types. Consistent with this, we found that
chemogenetic recruitment of PV interneurons suppressed convulsive behaviors by more than 80%. Our findings support the idea that selective chemogenetic enhancement of inhibitory synaptic pathways offers potential as an anti-seizure strategy.
PDF
