Excitatory Action Research Articles

Chronic VEGFR-3 signaling preserves dendritic arborization and sensitization under stress

Dendritic arborization is critical for the establishment and maintenance of precise neural circuits. Vascular endothelial growth factor D (VEGF-D), well-characterized as a “lymphangiogenic” growth factor, reportedly maintains dendritic arborization and synaptic strength in the hippocampus of adult mice through VEGF receptor (VEGFR-3) signaling. Here, we investigated the effect of chronic VEGFR-3-specific activation on adipose arbor morphometry using the Adipo-VD mouse, a model of inducible, adipose-specific VEGF-D overexpression. We examined whether adipose tissue innervation was preserved or functionally different in Adipo-VD mice during stress in vivo and if VEGFR-3 signaling afforded neuroprotection to challenged neurons in vitro. Chronic VEGFR-3 signaling in Adipo-VD subcutaneous adipose tissue resulted in a reduction in the dendrite length, dendritic terminal branches (filament length), and dendritic terminal branch volume (filament volume), but increased dendrite branching. We also identified reduced stimulus-evoked excitatory sympathetic nerve activity in Adipo-VD mice. Following 6-hydroxydopamine (6-OHDA) denervation, Adipo-VD dendritic arbors were preserved, including improved dendritic branch volume, length, and dendritic branches than in wildtype tissues. In vitro, we found that chronic elevation of VEGFR-3 signaling in developing mVC neurons changes the dendritic arbor complexity and improves stress-induced structure remodeling. Developing neurons are conferred neuroprotection against stress, potentially by upregulation of proteolytic conversion of pro-BDNF to mature BDNF. Mature neurons, however, display improved dendritic arbor complexity, and unaltered dendritic structural remodeling and improved resistance to stress with VEGFR-3 signaling. Overall, chronically increasing VEGFR-3 signaling in neurons has a synergistic impact on neurosensitization and neuroprotection during stress.

Analysis of the goals scored in the 2016 and 2017 CONMEBOL Libertadores Cups

ABSTRACT This study aimed to describe regularities in soccer goal-ending sequences in the 2016 and 2017 CONMEBOL Libertadores Cups. Observational methodology was used. The sample comprised 699 attack sequences that ended in goal from 250 games played in 2016 and 2017. An ad hoc observational instrument was designed. Data analysis was performed in three complementary ways: sequential, graph theory, and polar coordinates. Two sequences were highlighted: Pass with the head-Shot-Goal and Rebound-Rebound-Shot-Goal. The foot pass action is the most common prior to a shot. In addition, dribbling and conducting were excitatory and inhibitory actions, respectively, to score a goal. In conclusion, the use of open game was relevant to score goals and different patterns were observed.

Kv1.1 channels mediate network excitability and feed-forward inhibition in local amygdala circuits

Kv1.1 containing potassium channels play crucial roles towards dampening neuronal excitability. Mice lacking Kv1.1 subunits (Kcna1−/−) display recurrent spontaneous seizures and often exhibit sudden unexpected death. Seizures in Kcna1−/− mice resemble those in well-characterized models of temporal lobe epilepsy known to involve limbic brain regions and spontaneous seizures result in enhanced cFos expression and neuronal death in the amygdala. Yet, the functional alterations leading to amygdala hyperexcitability have not been identified. In this study, we used Kcna1−/− mice to examine the contributions of Kv1.1 subunits to excitability in neuronal subtypes from basolateral (BLA) and central lateral (CeL) amygdala known to exhibit distinct firing patterns. We also analyzed synaptic transmission properties in an amygdala local circuit predicted to be involved in epilepsy-related comorbidities. Our data implicate Kv1.1 subunits in controlling spontaneous excitatory synaptic activity in BLA pyramidal neurons. In the CeL, Kv1.1 loss enhances intrinsic excitability and impairs inhibitory synaptic transmission, notably resulting in dysfunction of feed-forward inhibition, a critical mechanism for controlling spike timing. Overall, we find inhibitory control of CeL interneurons is reduced in Kcna1−/− mice suggesting that basal inhibitory network functioning is less able to prevent recurrent hyperexcitation related to seizures.

Asymmetry is central to excitatory glutamate receptor activation.

Asymmetry is central to excitatory glutamate receptor activation.

IL-33/ST2 Signaling Regulates Synaptic Plasticity and Homeostasis in the Adult Hippocampal Circuitry.

In response to neuronal activity changes, the adult hippocampal circuits undergo continuous synaptic remodeling, which is essential for information processing, learning, and memory encoding. Glial cells, including astrocytes and microglia, actively regulate hippocampal synaptic plasticity by coordinating the neuronal activity-induced synaptic changes at the circuit level. Emerging evidence suggests that the crosstalk between neurons and glia in the adult hippocampus is region specific and that the mechanisms controlling this process are critically dependent on secreted factors. Interleukin-33 (IL-33), a cytokine of the IL-1 family, is a key factor that modulates such glia-driven neuromodulations in two distinct hippocampal circuits. The activation of IL-33 and its receptor complex is important for maintaining the excitatory synaptic activity in the cornu ammonis 1 subregion and the remodeling of dentate gyrus synapses through activity-dependent astrocyte-synapse and microglia-synapse interactions, respectively. Meanwhile, the dysregulation of this signaling is implicated in multiple neurological disorders, especially Alzheimer's disease. Further investigations of how IL-33/ST2 signaling is regulated in a region-specific manner as well as its diverse functions in glia-synapse communications in the adult hippocampal circuitry will provide insights into the nature of hippocampal synaptic plasticity and homeostasis in health and disease.

Presynaptic GABAB receptor\u2013mediated network excitation in the medial prefrontal cortex of Tsc2+/-\u2009mice

The TSC1 and TSC2 tumor suppressor genes control the activity of mechanistic target of rapamycin (mTOR) pathway. Elevated activity of this pathway in Tsc2+/- mouse model leads to reduction of postsynaptic GABAB receptor–mediated inhibition and hyperexcitability in the medial prefrontal cortex (mPFC). In this study, we asked whether presynaptic GABAB receptors (GABABRs) can compensate this shift of hyperexcitability. Experiments were performed in brain slices from adolescent wild-type (WT) and Tsc2+/- mice. Miniature and spontaneous postsynaptic currents (m/sPSCs) were recorded from layer 2/3 pyramidal neurons in mPFC using patch-clamp technique using a Cs+-based intrapipette solution. Presynaptic GABABRs were activated by baclofen (10 µM) or blocked by CGP55845 (1 µM). Independent on genotype, GABABR modulators bidirectionally change miniature excitatory postsynaptic current (mEPSC) frequency by about 10%, indicating presynaptic GABABR-mediated effects on glutamatergic transmission are comparable in both genotypes. In contrast, frequencies of both mIPSCs and sIPCSs were suppressed by baclofen stronger in Tsc2+/- neurons than in WT ones, whereas CGP55845 significantly increased (m/s)IPSC frequencies only in WT cells. Effects of baclofen and CGP55845 on the amplitudes of evoked (e)IPSCs confirmed these observations. These data indicate (1) that GABAergic synapses are inhibited by ambient GABA in WT but not in Tsc2+/- slices, and (2) that baclofen shifts the E/I ratio, determined as the ratio of (m/s)EPSC frequency to (m/s)IPSC frequency, towards excitation only in Tsc2+/- cells. This excitatory presynaptic GABABR-mediated action has to be taken into account for a possible medication of mental disorders using baclofen.Graphical abstract

Knock-in models related to Alzheimer\u2019s disease: synaptic transmission, plaques and the role of microglia

BackgroundMicroglia are active modulators of Alzheimer’s disease but their role in relation to amyloid plaques and synaptic changes due to rising amyloid beta is unclear. We add novel findings concerning these relationships and investigate which of our previously reported results from transgenic mice can be validated in knock-in mice, in which overexpression and other artefacts of transgenic technology are avoided.MethodsAppNL-F and AppNL-G-F knock-in mice expressing humanised amyloid beta with mutations in App that cause familial Alzheimer’s disease were compared to wild type mice throughout life. In vitro approaches were used to understand microglial alterations at the genetic and protein levels and synaptic function and plasticity in CA1 hippocampal neurones, each in relationship to both age and stage of amyloid beta pathology. The contribution of microglia to neuronal function was further investigated by ablating microglia with CSF1R inhibitor PLX5622.ResultsBoth App knock-in lines showed increased glutamate release probability prior to detection of plaques. Consistent with results in transgenic mice, this persisted throughout life in AppNL-F mice but was not evident in AppNL-G-F with sparse plaques. Unlike transgenic mice, loss of spontaneous excitatory activity only occurred at the latest stages, while no change could be detected in spontaneous inhibitory synaptic transmission or magnitude of long-term potentiation. Also, in contrast to transgenic mice, the microglial response in both App knock-in lines was delayed until a moderate plaque load developed. Surviving PLX5266-depleted microglia tended to be CD68-positive. Partial microglial ablation led to aged but not young wild type animals mimicking the increased glutamate release probability in App knock-ins and exacerbated the App knock-in phenotype. Complete ablation was less effective in altering synaptic function, while neither treatment altered plaque load.ConclusionsIncreased glutamate release probability is similar across knock-in and transgenic mouse models of Alzheimer’s disease, likely reflecting acute physiological effects of soluble amyloid beta. Microglia respond later to increased amyloid beta levels by proliferating and upregulating Cd68 and Trem2. Partial depletion of microglia suggests that, in wild type mice, alteration of surviving phagocytic microglia, rather than microglial loss, drives age-dependent effects on glutamate release that become exacerbated in Alzheimer’s disease.

EEGs Disclose Significant Brain Activity Correlated with Synaptic Fickleness.

Simple SummaryIn this study, we explore the emergence of oscillatory behavior similar to the signals of brain activity observed in electroencephalograms (EEGs) using a network of synaptic relations mingling excitatory and inhibitory neuron nodes. We identify abrupt variations on that activity brought about by swift synaptic mediations. These changes are originated by the slowdown of the activity of inhibitory neuron populations due to synaptic depression. The latter generates an imbalance between excitation and inhibition causing a quick explosive increase of excitatory activity, which turns out to be a (first-order) phase transition among different oscillatory states. Interestingly enough, near this transition, our model system exhibits oscillatory activity with a strong component in the delta-theta domain that coexist with fast oscillations and happens to be similar to the observed delta-gamma and theta-gamma modulation in actual brains. Our findings here help to understand actual brain activity data in terms of nonequilibrium phase transitions theory.We here study a network of synaptic relations mingling excitatory and inhibitory neuron nodes that displays oscillations quite similar to electroencephalogram (EEG) brain waves, and identify abrupt variations brought about by swift synaptic mediations. We thus conclude that corresponding changes in EEG series surely come from the slowdown of the activity in neuron populations due to synaptic restrictions. The latter happens to generate an imbalance between excitation and inhibition causing a quick explosive increase of excitatory activity, which turns out to be a (first-order) transition among dynamic mental phases. Moreover, near this phase transition, our model system exhibits waves with a strong component in the so-called delta-theta domain that coexist with fast oscillations. These findings provide a simple explanation for the observed delta-gamma and theta-gamma modulation in actual brains, and open a serious and versatile path to understand deeply large amounts of apparently erratic, easily accessible brain data.

Role of hydrogen sulfide in ventilatory responses to hypercapnia in the medullary raphe of adult rats.

What is the central question of this study? There is evidence that H2 S plays a role in the control of breathing: what are its actions on the ventilatory and thermoregulatory responses to hypercapnia via effects in the medullary raphe, a brainstem region that participates in the ventilatory adjustments to hypercapnia? What is the main finding and its importance? Hypercapnia increased the endogenous production of H2 S in the medullary raphe. Inhibition of endogenous H2 S attenuated the ventilatory response to hypercapnia in unanaesthetized rats, suggesting its excitatory action via the cystathionine β-synthase-H2 S pathway in the medullary raphe. Hydrogen sulfide (H2 S) has been recently recognized as a gasotransmitter alongside carbon monoxide (CO) and nitric oxide (NO). H2 S seems to modulate the ventilatory and thermoregulatory responses to hypoxia and hypercapnia. However, the action of the H2 S in the medullary raphe (MR) on the ventilatory responses to hypercapnia remains to be elucidated. The present study aimed to assess the role of H2 S in the MR (a brainstem region that contains CO2 -sensitive cells and participates in the ventilatory adjustments to hypercapnia) in the ventilatory responses to hypercapnia in adult unanaesthetized Wistar rats. To do so, aminooxyacetic acid (AOA; a cystathionine β-synthase (CBS) enzyme inhibitor), propargylglycine (PAG; a cystathionine γ-lyase enzyme inhibitor) and sodium sulfide (Na2 S; an H2 S donor) were microinjected into the MR. Respiratory frequency (fR ), tidal volume (VT ), ventilation ( ), oxygen consumption ( ) and body temperature (Tb ) were measured under normocapnic (room air) and hypercapnic (7% CO2 ) conditions. H2 S concentration within the MR was determined. Microinjection of the drugs did not affect fR , VT and during normocapnia when compared to the control group. However, the microinjection of AOA, but not PAG, attenuated fR and during hypercapnia in comparison to the vehicle group, but had no effects on Tb . In addition, we observed an increase in the endogenous production of H2 S in the MR during hypercapnia. Our findings indicate that endogenously produced H2 S in the MR plays an excitatory role in the ventilatory response to hypercapnia, acting through the CBS-H2 S pathway.

Differential regulation of excitatory synaptic transmission in the hippocampus and anterior temporal lobe by cyclin dependent kinase 5 (Cdk5) in mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS)

Mesial temporal lobe epilepsy with hippocamapal sclerosis (MTLE-HS) is the most common form of drug resistant epilepsy (DRE). MTLE-HS is a distributed network disorder comprising of not only the hippocampus, but other anatomically related extrahippocampal regions. Excitatory synaptic transmission is differentially regulated in the hippocampal and extra-hippocampal regions of patients with MTLE-HS, but its mechanism not understood. Cyclin-dependent kinase 5 (Cdk5) is known to regulate synaptic transmission and plasticity through up-regulation of NMDA receptors by phosphorylating NR2Asubunits. The present study is designed to investigate whether Cdk5 differentially regulates the excitatory synaptic transmission in the hippocampus and anterior temporal lobe (ATL) samples obtained from patients of MTLE-HS. We have measured the Cdk5 kinase activity and the protein levels of Cdk5, p-Cdk5, p35/p25, NR2A, pNR2A in the hippocampal and ATL samples obtained from patients with MTLE-HS. We have also determined the effect of roscovitine, a Cdk5 antagonist, on spontaneous excitatory postsynaptic currents (EPSCs) recorded from the hippocampal and ATL using patch-clamp technique. We observed significant increase in the expression of Cdk5, p-Cdk5, p35/p25, NR2A, pNR2A in the ATL samples as compared to the hippocampal samples. Cdk5 activity was significantly higher in ATL samples as compared to the hippocampal samples. Magnitude of reduction in the frequency of EPSCs by roscovitine in the ATL samples was higher than that in the hippocampal samples. Our studies suggest that Cdk5 differentially regulates excitatory synaptic activity in the hippocampal and ATL region of patients with MTLE-HS.

Mechanical Principles Governing the Shapes of Dendritic Spines.

Dendritic spines are small, bulbous protrusions along the dendrites of neurons and are sites of excitatory postsynaptic activity. The morphology of spines has been implicated in their function in synaptic plasticity and their shapes have been well-characterized, but the potential mechanics underlying their shape development and maintenance have not yet been fully understood. In this work, we explore the mechanical principles that could underlie specific shapes using a minimal biophysical model of membrane-actin interactions. Using this model, we first identify the possible force regimes that give rise to the classic spine shapes—stubby, filopodia, thin, and mushroom-shaped spines. We also use this model to investigate how the spine neck might be stabilized using periodic rings of actin or associated proteins. Finally, we use this model to predict that the cooperation between force generation and ring structures can regulate the energy landscape of spine shapes across a wide range of tensions. Thus, our study provides insights into how mechanical aspects of actin-mediated force generation and tension can play critical roles in spine shape maintenance.

Neuritin improves the neurological functional recovery after experimental intracerebral hemorrhage in mice

Stroke is one of the leading causes of death worldwide, with intracerebral hemorrhage (ICH) being the most lethal subtype. Neuritin (Nrn) is a neurotropic factor that has been reported to have neuroprotective effects in acute brain and spinal cord injury. However, whether Nrn has a protective role in ICH has not been investigated. In this study, ICH was induced in C57BL/6 J mice by injection of collagenase VII, while the overexpression of Nrn in the striatum was induced by an adeno-associated virus serotype 9 (AAV9) vector. We found that compared with GFP-ICH mice, Nrn-ICH mice showed improved performance in the corner, cylinder and forelimb tests after ICH, and showed less weight loss and more rapid weight recovery. Overexpression of Nrn reduced brain lesions, edema, neuronal death and white matter and synaptic integrity dysfunction caused by ICH. Western blot results showed that phosphorylated PERK and ATF4 were significantly inhibited, while phosphorylation of Akt/mammalian target of rapamycin was increased in the Nrn-ICH group, compared with the GFP-ICH group. Whole cell recording from motor neurons indicated that overexpression of Nrn reversed the decrease of spontaneous excitatory postsynaptic currents (sEPSCs) and action potential frequencies induced by ICH. These data show that Nrn improves neurological deficits in mice with ICH by reducing brain lesions and edema, inhibiting neuronal death, and possibly by increasing neuronal connections.

Clinical Signs and Pathology Associated With Domoic Acid Toxicosis in Southern Sea Otters (Enhydra lutris nereis)

The marine biotoxin domoic acid (DA) is an analog of the neurotransmitter glutamate that exerts potent excitatory activity in the brain, heart, and other tissues. Produced by the diatomPseudo-nitzschiaspp., DA accumulates in marine invertebrates, fish, and sediment. Southern sea otters (Enhydra lutris nereis) feed primarily on invertebrates, including crabs and bivalves, that concentrate and slowly depurate DA. Due to their high prey consumption (25% of body weight/day), sea otters are commonly exposed to DA. A total of 823 necropsied southern sea otters were examined to complete this study; first we assessed 560 subadult, adult, and aged adult southern sea otters sampled from 1998 through 2012 for DA-associated pathology, focusing mainly on the central nervous system (CNS) and cardiovascular system. We applied what was learned to an additional cohort of necropsied sea otters of all demographics (including fetuses, pups, juveniles, and otters examined after 2012:n= 263 additional animals). Key findings derived from our initial efforts were consistently observed in this more demographically diverse cohort. Finally, we assessed the chronicity of DA-associated pathology in the CNS and heart independently for 54 adult and aged adult sea otters. Our goals were to compare the temporal consistency of DA-associated CNS and cardiovascular lesions and determine whether multiple episodes of DA toxicosis could be detected on histopathology. Sea otters with acute, fatal DA toxicosis typically presented with neurological signs and severe, diffuse congestion and multifocal microscopic hemorrhages (microhemorrhages) in the brain, spinal cord, cardiovascular system, and eyes. The congestion and microhemorrhages were associated with detection of high concentrations of DA in postmortem urine or gastrointestinal content and preceded histological detection of cellular necrosis or apoptosis. Cases of chronic DA toxicosis often presented with cardiovascular pathology that was more severe than the CNS pathology; however, the lesions at both sites were relatively quiescent, reflecting previous damage. Sea otters with fatal subacute DA toxicosis exhibited concurrent CNS and cardiovascular pathology that was characterized by progressive lesion expansion and host response to DA-associated tissue damage. Acute, subacute, and chronic cases had the same lesion distribution in the CNS and heart. CNS pathology was common in the hippocampus, olfactory, entorhinal and parahippocampal cortex, periventricular neuropil, and ventricles. The circumventricular organs were identified as important DA targets; microscopic examination of the pituitary gland, area postrema, other circumventricular organs, and both eyes facilitated confirmation of acute DA toxicosis in sea otters. DA-associated histopathology was also common in cardiomyocytes and coronary arterioles, especially in the left ventricular free wall, papillary muscles, cardiac apex, and atrial free walls. Progressive cardiomyocyte loss and arteriosclerosis occurred in the same areas, suggesting a common underlying mechanism. The temporal stage of DA-associated CNS pathology matched the DA-associated cardiac pathology in 87% (n= 47/54) of cases assessed for chronicity, suggesting that the same underlying process (e.g., DA toxicosis) was the cause of these lesions. This temporally matched pattern is also indicative of a single episode of DA toxicosis. The other 13% of examined otters (n= 7/54) exhibited overlapping acute, subacute, or chronic DA pathology in the CNS and heart, suggestive of recurrent DA toxicosis. This is the first rigorous case definition to facilitate diagnosis of DA toxicosis in sea otters. Diagnosing this common but often occult condition is important for improving clinical care and assessing population-level impacts of DA exposure in this federally listed threatened subspecies. Because the most likely source of toxin is through prey consumption, and because humans, sea otters, and other animals consume the same marine foods, our efforts to characterize health effects of DA exposure in southern sea otters can provide strong collateral benefits.

Staving Off the Heat With Microglia.

Microglial Displacement of GABAergic Synapses Is a Protective Event During Complex Febrile Seizures Wan Y, Feng B, You Y, et al. Cell Reports. 2020;33(5):108346. doi:10.1016/j.celrep.2020.108346Complex febrile seizures (FSs) lead to a high risk of intractable temporal lobe epilepsy during adulthood, yet the pathological process of complex FSs is largely unknown. Here, we demonstrate that activated microglia extensively associated with glutamatergic neuronal soma displace surrounding GABAergic presynapses in complex FSs. Patch clamp electrophysiology establishes that the microglial displacement of GABAergic presynapses abrogates a complex-FS-induced increase in GABAergic neurotransmission and neuronal excitability, whereas GABA exerts an excitatory action in this immature stage. Pharmacological inhibition of microglial displacement of GABAergic presynapses or selective ablation of microglia in CD11bDTR mice promotes the generation of complex FSs. Blocking or deleting the P2Y12 receptor (P2Y12R) reduces microglial displacement of GABAergic presynapses and shortens the latency of complex FSs. Together, microglial displacement of GABAergic presynapses, regulated by P2Y12R, reduces neuronal excitability to mitigate the generation of complex FSs. Microglial displacement is a protective event during the pathological process of complex FSs.

Regulation of stress-provoked aggressive behavior using endocannabinoids

Reactive impulsive aggression is characterized by outbursts of rage and violence when subjects encounter threatening stressful events. Although impulsive aggression and violence create a high-cost burden on health and society, relatively little is known about treatment. Early adolescent social isolation (SI) alters brain development and functions. It induces hyper-excitatory in the ventral hippocampus (vHip) to promote acute stress-provoked outbursts of aggression, referred to as impulsive aggression, in mouse models. Cannabinoid type 1 receptors (CB1Rs) act on presynaptic sites and suppress neurotransmitter release into synapses. Given that CB1R activation inhibits neurotransmitter releases and modulates excitatory network activity, we tested the hypothesis that CB1R activation reduces impulsive aggression in SI mice through decreasing excitatory activity in the vHip. Here, we report that CB1R agonists, WIN-552122 (WIN) or arachidonylcyclopropylamide (ACPA), ameliorated acute stress-provoked attack behavior in the resident-intruder test without affecting general locomotion activity. Increasing endocannabinoids (eCBs) by inhibiting degradation enzymes in the vHip reduced impulsive aggression, and the effect was blunted by administration of AM251, a CB1R antagonist. Acute stress in SI mice induced c-Fos expression, a marker of neuronal activation, on vHip neurons projecting to the ventromedial hypothalamus (VMH), a well-known brain area that controls attack behavior. eCB augmentation inhibited c-Fos expression in VMH-projecting vHip neurons surrounded by CB1Rs. These results suggest that enhancing eCB signaling in order to activate CB1Rs suppresses impulsive aggression via suppressing vHip→VMH neural activity and point to a role of CB1R activation in ameliorating impulsive aggression in adults who have had adverse experiences during early adolescence.

5‐HT1A Receptor Activation by Buspirone Facilitates Post‐Inflammatory Intestinal Hypersensitivity in a Rat Model

Intestinal hypersensitivity is the most common symptom and cause of pain in patients with post-inflammatory irritable bowel syndrome (PI-IBS). Several lines of evidence suggest that central serotonin (5-HT) and its receptors significantly contribute to the phenomenon and therefore may be prospective therapeutic targets for relieving abdominal pain and discomfort in PI-IBS. We demonstrated previously that buspirone, a partial agonist of the 5-HT1A receptors, dose-dependently inhibited visceromotor reactions and medullary neuronal responses to noxious colorectal distension in healthy awake dogs and anesthetized rats, indicating the drug's antinociceptive properties in normal conditions. However, whether the 5-HT1A agonist can be beneficial for post-inflammatory intestinal hypersensitivity and hyperalgesia, which are thought to be associated with altered central 5-HT signaling, is still unknown. In the presented study, the effect of systemic buspirone (Sigma-Aldrich, USA) on intestinal hypersensitivity and its underlying neuronal mechanisms were evaluated in Wistar rats recovered from trinitrobenzene sulfonic acid (TNBS)-induced colitis. To assess gut sensitivity, the electromyography recordings of visceromotor reaction to colorectal distension (CRD) and extracellular microelectrode recordings of CRD-evoked caudal medulla neuronal responses were performed in conscious and urethane-anesthetized animals respectively. The results showed that in intestinal hypersensitive rats the visceromotor reaction and medullary neuronal responses to CRD in 40-60 days after TNBS treatment, despite the absence of obvious gut damage by this time, were increased compared with those in norm. Subcutaneous injection of buspirone (2 mg/kg), which exerted antinociceptive effect in healthy control group, further enhanced visceromotor reaction to CRD in post-colitis animals. In neurophysiological experiments on intestinal hypersensitivity rats, activation of the 5-HT1A receptors by intravenously administered buspirone (2 and 4 mg/kg) resulted in a dose-dependent increase of the CRD-evoked activity in the majority of the caudal medullary neurons, suggesting that the pro-algesic effect of the drug can at least partially be mediated by its excitatory action at the supraspinal level. These data demonstrate that the 5-HT1A agonist buspirone loses its antinociceptive properties in post-colitis conditions and thus may not be useful for reliving visceral hypersensitivity and hyperalgesia in PI-IBS.

Astroglial Regulation of Caloric Intake Following Acute High‐Fat Diet Exposure

Chronic high fat diet (HFD) exposure results in hyperphagia and the development of obesity. Caloric intake during acute (3-5 days) HFD exposure, however, is regulated. We have previously shown that increased activation of NMDA receptors on the motoneurons of the dorsal motor nucleus of the vagus (DMV) contributes to the regulation of caloric intake during acute HFD exposure (Clyburn, et. al. 2018). The mechanism that is responsible for altered excitatory drive to DMV neurons is unknown. HFD exposure is associated with inflammation. Neuroinflammation and astroglial activation have been shown to modulate neural activity and glutamatergic signaling. The purpose of this study is to test the hypothesis that during acute HFD exposure, astrocyte activation increases the glutamatergic drive to DMV neurons and regulates caloric intake. Male and female Sprague-Dawley rats were transfected with the astrocyte inhibitory DREADD, pAAV-GFAP-hM4D(Gi)-mCherry, (“hM4Di”; Addgene #50479-AAV5; titer 1x1013 diluted 1:10) or AAV-GFAP104-mCherry, (“empty vector”; UNC vector core, titer 2.7x1012 diluted 1:10). Microinjections were made into the dorsal vagal complex. After recovery, rats were fed a control or HFD (14% and 60% kcal from fat, respectively) for 3-5 days. Whole-cell patch clamp recordings were made of gastric-projecting DMV neurons in thin brainstem slices of transfected rats. The effects of the NMDA receptor antagonist, AP5 (25μM), in the presence of clozapine (10μM) or the DREADD agonist, clozapine-N-oxide (CNO; 10μM) on miniature excitatory postsynaptic current (mEPSC) kinetics and action potential (AP) firing rate were examined. The effects of CNO and clozapine on caloric intake and gastric emptying rates using the 13C octanoic acid breath test were also assessed. As shown previously, following acute HFD, AP5 decreases mEPSC frequency and AP firing rate. This AP5-mediated decrease was unaffected by clozapine or CNO application to empty vector control groups. CNO in the hM4Di group attenuated the AP5-mediated decrease in mEPSC frequency and AP firing rate (mEPSC frequency: 54±5.0% vs. 53±9.5% vs. 100±1.5% of baseline, respectively; AP firing rate: 8±5.3% vs. 41±14.9% vs. 111±3.5%; P<0.05 with two-way ANOVA and post-hoc Bonferroni; N=7-9 cells from 3-5 rats). In addition, the delay in gastric emptying rates observed 4 days following HFD exposure was attenuated by CNO administration to hM4Di rats, while empty vector or clozapine controls were uneffected (119±7.9% vs. 166±7.4% vs. 144±6.7% of baseline, respectively; P<0.05 with two-way ANOVA and post-hoc Bonferroni; N=6-7 rats). CNO administration to hM4Di rats also prevented the regulation of caloric intake normally observed 3-5 days of HFD exposure, compared to clozapine and empty vector controls (AUC: 964±34.4a.u. vs. 843±14.3a.u. vs. 815±17.9a.u., respectively; P<0.05 from Student's unpaired t-test; N=6). These data suggest that astrocyte activation is involved in the alteration of glutamatergic currents and the homeostatic regulation of food intake following acute HFD. Understanding the mechanism responsible for the regulation of caloric intake is critical in identifying potential therapeutic targets for the treatment of obesity when such neuroplasticity may be absent.

The Effects of In Vitro Seizure‐Like Activity on Actin Capping in Dendritic Spines

Neonatal seizures can cause dysregulation of excitatory synaptic activity, that may contribute to the development of cognitive deficits later in life. While the mechanisms underlying this association are not known, one possibility is the regulation of dendritic spines, the postsynaptic structures of most of the excitatory synapses in the mammalian brain. These membranous protrusions function as chemical and electrical micro-compartments, transducing individual signals of excitatory neurotransmitters. Structural plasticity, the ability of spines to change their shape and thus their function in response to synaptic activity, underlies learning and memory, and many neurological diseases defined by altered cognitive abilities display dendritic spine dysgenesis. This shape change requires regulation of actin mesh-works by actin binding proteins (ABPs) within dendritic spines. Decreased concentrations of functional ABPs induces abnormalities in the density, morphology, and dynamic nature of dendritic spines, resulting in changes in synaptic function such as hindered synaptic signal amplitude. Based upon these studies, we hypothesized that seizure activity causes deficits in the structural plasticity of dendritic spines via depletion of functional ABPs. To investigate the molecular physiological consequences of uncontrolled, synaptic excitability on intracellular actin regulation, we used a zero magnesium model to induce in vitro seizure-like activity in rat hippocampal neurons. At multiple time points after zero Mg2+ exposure, we assessed localization of ABPs in dendritic spines using immunocytochemistry and analyzed the motility of dendritic spines in response to excitatory stimulation using live-cell imaging of GFP-actin transfected neurons. Preliminary results show a neuronal age and time point dependent decrease in ABP localization and spine motility in dendritic spines after in vitro seizure-like activity. Taken together, these data indicate that seizure-like activity can alter ABPs and baseline spine dynamics. Ongoing work will examine whether seizure-induced ABPs could affect structural plasticity of dendritic spines to determine the feasibility of ABPs as a therapeutic target to decrease cognitive deficits that can result from early-life seizures.

Chronic lithium treatment alters the excitatory/ inhibitory balance of synaptic networks and reduces mGluR5-PKC signalling in mouse cortical neurons.

Background:Bipolar disorder is characterized by cyclical alternation between mania and depression, often comorbid with psychosis and suicide. Compared with other medications, the mood stabilizer lithium is the most effective treatment for the prevention of manic and depressive episodes. However, the pathophysiology of bipolar disorder and lithium’s mode of action are yet to be fully understood. Evidence suggests a change in the balance of excitatory and inhibitory activity, favouring excitation in bipolar disorder. In the present study, we sought to establish a holistic understanding of the neuronal consequences of lithium exposure in mouse cortical neurons, and to identify underlying mechanisms of action.Methods:We used a range of technical approaches to determine the effects of acute and chronic lithium treatment on mature mouse cortical neurons. We combined RNA screening and biochemical and electrophysiological approaches with confocal immunofluorescence and live-cell calcium imaging.Results:We found that only chronic lithium treatment significantly reduced intracellular calcium flux, specifically by activating metabotropic glutamatergic receptor 5. This was associated with altered phosphorylation of protein kinase C and glycogen synthase kinase 3, reduced neuronal excitability and several alterations to synapse function. Consequently, lithium treatment shifts the excitatory–inhibitory balance toward inhibition.Limitations:The mechanisms we identified should be validated in future by similar experiments in whole animals and human neurons.Conclusion:Together, the results revealed how lithium dampens neuronal excitability and the activity of the glutamatergic network, both of which are predicted to be overactive in the manic phase of bipolar disorder. Our working model of lithium action enables the development of targeted strategies to restore the balance of overactive networks, mimicking the therapeutic benefits of lithium but with reduced toxicity.

Ophn1 regulation of prefrontal inhibition: A mechanism for stress susceptibility in intellectual disability

Wang etal. (2021) characterize the molecular, cellular, and circuit-level role of Oligophrenin-1 in prefrontal parvalbumin interneurons, demonstrating that loss of Ophn1 function in these neurons is a mechanism for increased susceptibility to stress in intellectual disability caused by OPHN1 mutations.