NR2B Subunit Research Articles

Ketamine reduces remifentanil-induced postoperative hyperalgesia mediated by CaMKII-NMDAR in the primary somatosensory cerebral cortex region in mice

Remifentanil is commonly used clinically for perioperative pain relief, but it may induce postoperative hyperalgesia. Low doses of ketamine have remained a common choice in clinical practice, but the mechanisms of ketamine have not yet been fully elucidated. In this study, we examined the possible effects of ketamine on calcium/calmodulin-dependent protein kinase II α (CaMKIIα) and N-methyl-d-aspartate receptor (NMDAR) subunit NR2B in a mouse model of remifentanil-induced postoperative hyperalgesia (RIPH) in the primary somatosensory cerebral cortex (SI) region. The paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were used to assess mechanical allodynia and thermal hyperalgesia, respectively, before and after intraoperative remifentanil administration. Before surgery, mice received intrathecal injections of the following drugs: ketamine, NMDA, BayK8644 (CaMKII activator), and KN93 (CaMKII inhibitor). Immunofluorescence was performed to determine the anatomical location and expression of activated CaMKIIα, phosphorylated CaMKIIα (p-CaMKIIα). Additionally, western blotting was performed to assess p-CaMKIIα and NMDAR expression levels in the SI region. Remifentanil decreased the PWMT and PWTL at 0.5 h, 2 h, and 5 h and increased p-CaMKIIα expression in the SI region. Ketamine increased the PWMT and PWTL and reversed the p-CaMKIIα upregulation. Both BayK8644 and NMDA reversed the effect of ketamine, decreased the PWMT and PWTL, and upregulated p-CaMKIIα expression. In contrast, KN93 enhanced the effect of ketamine by reducing hyperalgesia and downregulating p-CaMKIIα expression. These results suggested that ketamine reversed RIPH by inhibiting the phosphorylation of CaMKIIα and the NMDA receptor in the SI region in mice.

Prenatal melamine exposure impairs cognitive flexibility and hippocampal synaptic plasticity in adolescent and adult female rats

Prenatal melamine exposure (PME) affects spatial cognition in adolescent male rats and impairs synaptic functions. Strikingly, these effects can persist into adulthood. The current experiments examined whether PME-induced behavioral defects would be observed in female offspring, and how these effects varied with age (adolescent and adult). After female rats were exposed to melamine through their entire gestational period (GD), their spatial cognition was tested using water maze tasks at postnatal day 36 (PD36) and PD90. Long-term potentiation (LTP) and long-term depression (LTD) were recorded from the hippocampal Schaffer collaterals to the CA1 area. Results indicated that PME led to substantial reversal learning deficits and disrupted LTD at both PD36 and PD90. Additionally, PME did not affect LTP, although a lower fEPSP slope was observed in the adolescent PME-treated group than in the adult PME-treated group. Additionally, PME did not induce a horizontal shift in the synaptic modification threshold but caused a downward shift of the frequency–response curve at the low-frequency stimulation (LFS) of 1.0 Hz. However, the paired-pulse facilitation (PPF) ratio and the input/output function were not affected. Although the expression of both NR1 and NR2B subunits of NMDA receptors were diminished at PD36, the NR1 level was not affected at PD90. These findings suggest that PME impairs spatial memory in adolescent and adult females, and the impairments of hippocampal synaptic function via the inhibition of NMDAR expression may play a role in these effects.

Involvement of brain-derived neurotrophic factor (BDNF) in the long-term memory effects of glucocorticoid stimulation during adolescence/young adulthood

Brain-derived neurotrophic factor (BDNF) has been implicated in cognition and the effects of chronic stress. We have previously shown in mice that chronic adolescent treatment with corticosterone (CORT), to simulate stress, resulted in spatial memory deficits and markedly elevated levels of the N-methyl-D-aspartate (NMDA) receptor subunit NR2B in adult male BDNF heterozygous mice (BDNF+/-), but not in wildtype controls (WT) or females. The aim of the present study was to further characterize this ‘two hit’ model, including whether these effects are long-lasting. CORT treatment was delivered in the drinking water from 6 to 9 weeks of age. As previously demonstrated, male BDNF+/− mice treated with CORT presented with a deficit in spatial memory at 11 weeks of age. However, this deficit was not maintained at 15 weeks of age. Conversely, male WT treated with CORT developed a deficit only at 15 weeks of age. There were no significant gene-environment interactions in female mice at any time point. CORT treatment caused a modest, but significant increase in NR2B levels which was independent of genotype. These results show marked age-dependent and sex-dependent effects of CORT on behaviour which are different in BDNF+/− mice than in controls.

Anterior insular cortex mediates hyperalgesia induced by chronic pancreatitis in rats

Central sensitization plays a pivotal role in the maintenance of chronic pain induced by chronic pancreatitis (CP), but cortical modulation of painful CP remains elusive. This study was designed to examine the role of anterior insular cortex (aIC) in the pathogenesis of hyperalgesia in a rat model of CP. CP was induced by intraductal administration of trinitrobenzene sulfonic acid (TNBS). Abdomen hyperalgesia and anxiety were assessed by von Frey filament and open field tests, respectively. Two weeks after surgery, the activation of aIC was indicated by FOS immunohistochemical staining and electrophysiological recordings. Expressions of VGluT1, NMDAR subunit NR2B and AMPAR subunit GluR1 were analyzed by immunoblottings. The regulatory roles of aIC in hyperalgesia and pain-related anxiety were detected via pharmacological approach and chemogenetics in CP rats. Our results showed that TNBS treatment resulted in long-term hyperalgesia and anxiety-like behavior in rats. CP rats exhibited increased FOS expression and potentiated excitatory synaptic transmission within aIC. CP rats also showed up-regulated expression of VGluT1, and increased membrane trafficking and phosphorylation of NR2B and GluR1 within aIC. Blocking excitatory synaptic transmission significantly attenuated abdomen mechanical hyperalgesia. Specifically inhibiting the excitability of insular pyramidal cells reduced both abdomen hyperalgesia and pain-related anxiety. In conclusion, our findings emphasize a key role for aIC in hyperalgesia and anxiety of painful CP, providing a novel insight into cortical modulation of painful CP and shedding light on aIC as a potential target for neuromodulation interventions in the treatment of CP.

Delayed audiogenic seizure development in a genetic rat model is associated with overactivation of ERK1/2 and disturbances in glutamatergic signaling

Krushinsky-Molodkina (KM) rats genetically prone to audiogenic seizure are characterized by age-dependent expression of audiogenic seizures (AGS). It is known that the critical period of enhanced seizure susceptibility in rodents occurs at 2nd–3rd weeks of postnatal development. However, KM rats do not express AGS at this time-point, but start to demonstrate a stable AGS only after the age of 3 months. We hypothesized that this delay in AGS susceptibility in KM rats is genetically determined and may depend on some alterations in the development of the hippocampal glutamatergic system during the early postnatal period. We analyzed the expression and activity of seizure-related proteins, such as vesicular glutamate transporter 2 (VGLUT2), extracellular signal-regulated kinases 1 and 2 (ERK1/2), synapsin I, and NR2B subunit of the N-methyl-d-aspartate (NMDA) receptor (NR2B) in the hippocampus of KM rats during postnatal development. A significantly higher activity of ERK1/2 in KM rats was observed at 14th, 30th, and 60th days of postnatal development (P14, P30, P60) in comparison with control Wistar rats of the corresponding ages, while in adult (P120) KM rats it was at the same level with Wistar rats. Despite the increased activity of ERK1/2 at P14 and P30, the phosphorylation of synapsin I at Ser62/67 was significantly lower in the hippocampus of KM rats than in Wistar rats of the same ages; however, at P60 and P120, the phosphorylation of synapsin I was enhanced. Our data also revealed the increase of VGLUT2 and NR2B expression at P14, which dramatically decreased at the later stages. Our data indicate that a genetically determined increase in ERK1/2 kinase activity during postnatal ontogenesis in KM rats may be associated with the disturbances in synthesis and activity of the proteins, which are responsible for glutamatergic transmission in the KM rat hippocampus during the seizure susceptibility development.

Sleep Deprivation Disrupts Acquisition of Contextual Fear Extinction by Affecting Circadian Oscillation of Hippocampal-Infralimbic proBDNF.

Extensive evidence showed that mature brain-derived neurotrophic factor (mBDNF) levels displayed a circadian pattern. Circadian disruption, for example, sleep deprivation (SD), induced functional and behavioral deficits. However, compared with that of mature form, the biological role of the pro-peptide, proBDNF, was poorly understood. Here, we found that proBDNF was expressed under circadian rhythm in the ventral hippocampus (vHPC). SD rats exhibited deficits in acquisition of conditioned extinction and damped rhythmicity in vHPC proBDNF activity that were accompanied by SD between zeitgeber time (ZT)0 and ZT4, but not the late stage of sleep period. Furthermore, SD affected fear extinction through vHPC-IL proBDNF signaling, which was associated with NR2B subunits of NMDA receptors. More importantly, infusion of proBDNF could mitigate SD-induced abnormal neural activity, by suppressing the enhanced basal firing rate of IL-RS and elevating the depressed neural response that evoked by acquisition of conditioned extinction. Therefore, this finding provided the first evidence that circadian oscillation of vHPC proBDNF activity contributed to the effects of SD on acquisition of conditioned fear extinction, and suggested a new therapeutic target to reverse the cognitive deficits in sleep-related mental disorder, such as post-traumatic stress disorder (PTSD).

Social defeat-induced increase in the conditioned rewarding effects of cocaine: Role of CX3CL1

Social stress is associated with higher vulnerability to drug use, as it enhances the reinforcing effects of psychostimulants in rodents. Furthermore, continued or severe stress induces a proinflammatory state of microglial activation and augmented cytokine production. The aim of the present work was to evaluate the role of fractalkine [C-X3-C motif ligand 1 (CX3CL1)], an inflammatory chemokine, in the increased conditioned rewarding effects of cocaine in animals exposed to social defeat stress. In addition, we measured the signaling cascade pathway of CX3CL1 in the hippocampus (HPC) (including p-ERK/ERK, p-p38/p38 MAPK, p-p65/p65 NFκB and p-CREB/CREB ratios). The glutamate receptor subunits NR1, NR2B and GluA1 were also assessed. A total of 102 adult male C57BL/6 J wild-type (WT) and Cx3cr1 knockout (KO) mice were divided into different experimental groups according to stress condition (exploration or social defeat). Three weeks after the last social defeat, conditioned place preference (CPP) was induced by a subthreshold dose of cocaine (1 mg/kg). Brain tissue samples were taken 24 h after the CPP procedure to determine the levels of the proteins and transcription factors. Our results showed that, in WT animals, repeated social defeat (RSD) decreased CX3CL1 striatal levels without producing changes in the HPC. In addition, RSD induced an increase in the conditioned rewarding effects of cocaine, regardless of the genotype. After CPP induced by cocaine, defeated Cx3cr1-deficient mice showed a decrease in the p-p65/p65 NFκB and pCREB/CREB ratio in the HPC, and an increase in the hippocampal levels of CX3CL1 and p-p38/p38 MAPK relation. In all defeated mice, there was a decrease in the ionotropic glutamate receptor subunit NR1. In conclusion, these results suggest that the lack of CX3CL1/Cx3cr1 signaling under stress conditions induces changes in protein and transcription factors, indicating that CX3CL1 is needed to shield the response to social defeat.

TMEM25 modulates neuronal excitability and NMDA receptor subunit NR2B degradation.

The expression of the transmembrane protein 25 gene (Tmem25) is strongly influenced by glutamate ionotropic receptor kainate type subunit 4, and its function remains unknown. Here, we showed that TMEM25 was primarily localized to late endosomes in neurons. Electrophysiological experiments suggested that the effects of TMEM25 on neuronal excitability were likely mediated by N-methyl-d-aspartate receptors. TMEM25 affected the expression of the N-methyl-d-aspartate receptor NR2B subunit and interacted with NR2B, and both were colocalized to late endosome compartments. TMEM25 induced acidification changes in lysosome compartments and accelerated the degradation of NR2B. Furthermore, TMEM25 expression was decreased in brain tissues from patients with epilepsy and epileptic mice. TMEM25 overexpression attenuated the behavioral phenotypes of epileptic seizures, whereas TMEM25 downregulation exerted the opposite effect. These results provide some insights into TMEM25 biology in the brain and the functional relationship between TMEM25 and epilepsy.

Motivation, risk-taking and sensation seeking behavior in propofol anesthesia exposed peripubertal rats

Adolescent neurodevelopment confer vulnerability to the actions of treatments that produce adaptations in neurocircuitry underlying motivation, impulsivity and reward. Considering wide usage of a sedative-hypnotic agent propofol in clinical practice, we examined whether propofol is a challenging treatment for peripubertal brain. Motivation/hedonic behavior (sucrose preference test), approach/avoidance behavior (elevated plus maze test) and response to dissociative drug phencyclidine (PCP) were studied in peripubertal rats (the rodent model of periadolescence) after propofol anesthesia exposure (PAE). Neurodegeneration (Fluoro-Jade staining) and the expression of proteins (Western blot) involved in excitatory synaptic transmission and activity-dependent synaptic stabilization in the medial prefrontal cortex (mPFC) and striatum (components of motivation/reward circuitry; process both appetitive and aversive events) were examined as well. In peripubertal rats PAE produced 1) transient brain-region specific changes in the expression of N-methyl-d-aspartate (NMDA) receptor subunits NR2A and NR2B, PSD-95 and N-cadherin, without neurotoxicity, 2) hyperlocomotor response to PCP, 3) no changes in preference for palatable 1% sucrose solution and a decrease in food eaten, 4) preference for 20% sucrose solution without changes in food eaten, 5) stretch-attended postures and open arms entries in the elevated plus maze test. Overall, these novel findings show that PAE leaves transient synaptic trace recognized as early form of synaptic plasticity related to passive drug exposure in the brain systems implicated in motivation/reward, increases drug-responsiveness, favors risk-taking and preference of novel/intense stimuli repairing otherwise present motivational deficiency. These findings accentuate multifaceted response to propofol in peripuberty and the importance of environmental stability for the most favorable neurobehavioral recovery.

Effects of Riluzole on Nr2B subunits of NMDA receptors in rat migraine model

Migraine, a major public health problem, occurring due to the consequences of serial changes in the nerve ganglion leading to an imbalance in the excitation and inhibition. The is the major neurotransmitter in the central nervous system, causing to the sensory neurons leading to sensitization and . The present study was done to determine the effects of on the Nr2B of NMDA receptors after inducing a migraine. The rats were treated with after inducing migraine with nitroglycerin 10mg/kg. The nitroglycerin treated rats showed intense staining for NR2B , and there was a decrease in the expression after treatment. This study concludes that the NR2B are during migraine and can be used to control those by its property.

Comprehensive QSAR studies reveal structural insights into the NR2B subtype selective benzazepine derivatives as N-Methyl-d-Aspartate receptor antagonists

NMDA receptors are considered as high profile therapeutic target in the treatment of pain and neurodegenerative diseases such as Alzheimer's, Huntington's and Parkinson's disease. NR2 subunit of NMDA receptor divided into four subunits i.e. NR2A, NR2B, NR2C and NR2D. Restricted distribution of NR2B subunit in the brain makes it potential target. In the present study, new structural insights into benzazepine derivatives as NR2B selective NMDA receptor antagonists have been reported using QSAR modelling. Total five QSAR models were developed using various statistical methods. 2D-QSAR models showed that chi3 and SsssNE-index descriptors are crucial for NMDA receptor antagonistic activity. Chi3 descriptor relates with non-hydrogen heteroatom connected with two or three bonds and SsssNE-index descriptor deals with Electrotopological state indices for number of nitrogen atom connected with three single bonds. Both the descriptors were negatively correlated with all the developed 2D-QSAR models. Significance of these descriptors is extensively studied in present work. In 3D-QSAR studies, steric and electrostatic fields were found to be important for antagonistic effect on NMDA receptors. 3D-QSAR studies clarify the substitution pattern of R group on nitrogen atom of benzazepine core. The substitutions with high electropositive with less or medium steric character are favourable for the activity. All developed models were critically validated to establish their reliability and accuracy for prediction of binding affinity of benzazepine derivatives towards NR2B subunit of NMDA receptor. The present study offered not only highly predictive and reliable QSAR models for benzazepine derivatives but also revealed some of the vital structural observations required for their higher binding affinity towards NR2B subunit of NMDA receptors.

Effects of enriched environment on the subunits N-mehyl-D-aspartate receptors at the 17th area of the visual cortex in adult amblyopia rats

Objective To explore the effect of enriched environment on the level of NR2A and NR2B subunits of N-mehyl-D-aspartate (NMDA) receptors which belong to glutamate receptors with excitability at the 17th area of the visual cortex in amblyopia rats after the critical period, and to understand the possible mechanism of synaptic plasticity of the visual cortex in adult amblyopia rats. Methods Eighty Wistar rats were divided into normal group and experimental group by random number table.Right eyelids of all rats were sutured through the whole critical period in order to establish monocular deprivation (MD) amblyopia model.The rats in experimental group were divided into the amblyopia group, standard environment (SE) group and environmental enrichment (EE) group on P45 in random.The sutured right eyelids were opened on P46 in the SE group and EE group.All rats were sacrificed to get the 17th area of the left visual cortex on P60, P75 and P105.Three rats were used at different time points from each group.The Ⅰ-Ⅵ layers of the visual cortex area 17 were observed by using hematoxylin-eosin staining.The expression of NMDA-NR2A and NMDA-NR2B was detected by immunohistochemistry.Integrated optical density of NMDA-NR2A and NMDA-NR2B was detected by using special image analysis software (Image-Pro Plus 6.0). The use of animals complied with Regulation on the Managenment Experimental Ainimals from Shandong Eye Institute and Association for Research in Vision and Ophthalmology (ARVO). Results The positive expression of NMDA-NR2A and NMDA-NR2B were observed in the visual cortex.The positive cells were mostly round or elliptical and mainly expressed in cell membrane.The expression of NMDA-NR2A in P60, P75 and P105 from four groups had statistical differences (all at P 0.05). Conclusions The plasticity of visual cortex exists not only in the critical period but also after the critical period of visual development.EE, as a non-invasion method, can improve and recover the synaptic plasticity in visual cortex of adult rats by the expression of NMDA-NR2A and NMDA-NR2B. Key words: Amblyopia; Visual cortex; Synapse; Enriched environment; N-mehyl-D-aspartate receptors

Superoxide generation via the NR2B-NMDAR/RasGRF1/NOX2 pathway promotes dendritogenesis.

N-methyl-D-aspartate receptors (NMDARs) that contain the NR2A and NR2B subunits play a critical role in neuronal plasticity and dendritogenesis. Gain-and-loss-of function studies indicate that NR2B, but not NR2A, promotes dendritic branching. Accumulating evidence indicates that stimulation of NMDARs activates NADPH oxidase (NOX2), thereby generating superoxide. However, the molecular underpinnings of this process are not understood. RasGRF1, a guanine nucleotide exchange factor, is key for several forms of neuronal plasticity and interacts directly with the tail of NR2B. We investigated whether the NR2B-NMDAR/RasGRF1 pathway regulates the activity of NOX2 and whether superoxide production is required for dendritogenesis. We measured superoxide production in developing primary cultures of hippocampal neurons from 3 to 25 days in vitro (DIV) with the probe dihydroethidium (dHE). We found the highest dHE levels at early and intermediate developmental stages (3-15 DIV), when the NR2B-NMDAR expression is abundant. During these early/intermediate developmental stages, but not in mature neurons (>15 DIV), NMDAR activity is required for superoxide production. We also found that disrupting the NR2B-RasGRF1 interaction led to reduced dHE fluorescence intensity and moreover inhibited dendritic branching in hippocampal neurons. Together, our data indicate that superoxide production is induced by the NR2B-NMDARs/RasGRF1/NOX2 pathway and promotes dendritogenesis.

Synthesis and Characterization in Rodent Brain of the Subtype-Selective NR2B NMDA Receptor Ligand [11C]Ro04-5595 as a Potential Radiotracer for Positron Emission Tomography.

The NR2B subunit of the N-methyl-d-aspartate (NMDA) receptor has been implicated in controlling synaptic plasticity, memory, and learning. Herein, we describe an 11C-labeled PET radiotracer based on 1-(4-chlorophenethyl)-6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolin-7-ol, Ro04-5595. The radiotracer was evaluated in rats using PET. The PET study showed a good pharmacokinetic profile with rapid uptake and washout over 90 min. Complementary high-resolution autoradiographic images using [3H]Ro04-5595 demonstrated strong binding in NR2B receptor-rich regions and low binding in cerebellum where NR2B concentration is low. We conclude to have developed a selective NR2B receptor radioligand suitable for quantitative and qualitative imaging of a NR2B receptor distribution in vitro and in vivo.

The positive allosteric modulation of GABAA receptors mRNA in immature hippocampal rat neurons by midazolam affects receptor expression and induces apoptosis

Background: Numerous experimental studies show that anesthetics are potentially toxic to the immature brain. Even though benzodiazepines are widely used in pediatric anesthesia and intensive care medicine, only a few studies examine the effects of these drugs on immature neurons.Methods: Hippocampal neuronal cell cultures of embryonic Wistar rats (15 days in culture) were incubated with midazolam 100 or 300 nM for either 30 min or 4 h. The time course of the mRNA expression of the glutamate receptors subunits NR1, NR2A and NR2B of the NMDA receptor, the GluA-1 and A-2 subunits of the AMPA receptor as well as the alpha 1 subunit of the GABAA receptor were examined by PCR. Apoptosis was detected using Western blot analysis for BAX, Bcl-2 and Caspase-3.Results: Midazolam at 100 and 300 nM applied for 30 min and 100 nM for 4 h affected glutamate receptor and GABAA receptor subunit expression. However, these effects were reversible within 72 h following washout. When 300 nM midazolam was applied for 4 h a significant increase in the NR 1 and NR 2A mRNA subunit expression could be detected. The increase in NR 2B receptor subunit expression as well as the GluA1 subunit expression was not reversible within 72 h following washout. This increase in mRNA glutamate receptor subunit expression was associated with a significant increase in neuronal apoptosis.Conclusion: In immature neurons midazolam altered GABA and glutamate mRNA receptor subunit expression. Prolonged increase in midazolam-induced glutamate receptor expression was associated with apoptosis.

Restraint devices for repetitive transcranial magnetic stimulation in mice and rats.

IntroductionRepetitive transcranial magnetic stimulation has been widely used for the treatment of neurological and psychiatric diseases. Rodent animals including mice and rats are often used to investigate the potential cellular and molecular mechanisms for the therapeutic effects of repetitive transcranial magnetic stimulation. So far there is no report about an easy‐to‐use device to restrain rodent animals for repetitive transcranial magnetic stimulation.Methods and ResultsWe introduced the design and use of the restraint device for mice or rats. In the mouse device, western blot and real‐time PCR analysis showed that，in stimulated mouse frontal cortex, 10 Hz high frequency stimulation for 10 sessions resulted in enhanced expression of NR2B‐containing N‐methyl‐D‐aspartic acid receptors and reduced α1 subunit of inhibitory GABAA receptors, whereas 0.5 Hz low frequency stimulation for 10 sessions caused decreased expression of NR2B subunit and increased α1 subunit of GABAA receptors. In the rat device, measures of motor evoke potentials indicated that 10 Hz stimulation for 10 sessions increased the excitability of stimulated cortex, whereas 0.5 Hz for 10 sessions reduced it.ConclusionsThese results suggested the effectiveness of the devices. Thus, the two devices are practical and easy‐to‐use to investigate the mechanisms of repetitive transcranial magnetic stimulation.

Implication of NMDA-NO pathway in the antidepressant-like effect of ellagic acid in male mice

Depression is one the common psychiatric disorders through the world. Nitric oxide (NO) and N-methyl-d-aspartate receptor (NMDA-R) are involved in the pathophysiology of depression. Previous studies have been reported various pharmacological properties for ellagic acid (EA). We aimed to evaluate possible involvement of NMDA-NO pathway in the antidepressant-like effect of EA. To do this, we used relevant behavioral tests to evaluate depressive-like behavior. In order to find effective and sub-effective doses of agents, mice treated with EA (6.25, 12.5, 25, 50 and 100 mg/kg), L-NAME (5 and 10 mg/kg), L-arg (25 and 50 mg/kg), NMDA (75 and 150 mg/kg) and ketamine (0.25 and 0.5 mg/kg). Furthermore, mice were treated with combination of sub-effective dose of EA plus sub-effective doses of L-NAME and/or ketamine as well as treated with effective dose of EA in combination of effective doses of L-arg and/or NMDA. Level of NO and gene expression of NR2A and NR2B subunits of NMDA-R were assessed in the hippocampus. Results showed that EA dose dependently provoked antidepressant-like effects and also decreased the hippocampal NO level as well as expression of NMDA-Rs. Co-administration of sub-effective doses of L-NAME or ketamine with sub-effective dose of EA potentiated the effect of EA on behaviors, NO level as well as NMDA-Rs gene expression in the hippocampus. However, co-treatment of effective dose of EA with effective doses of L-arg or NMDA mitigated effects of EA. In conclusion, our data suggested that NMDA-NO, partially at least, are involved in the antidepressant-like effect of EA.

The Role of Glutamatergic Antagonism in the Antidepressant‐like Activity of the Rapid‐Acting Antidepressant RO‐25‐6981 and Its Analogs

The glutamatergic antagonist, RO‐25‐6981, has previously been shown to exert both rapid and sustained antidepressant‐like activity. The purpose of the current study is to delineate its putative antidepressant mechanisms of NR2B‐selective NMDA receptor antagonism and monoamine reuptake transporter inhibition. RO‐25‐6981 and structural analogs with varying degrees of NR2B binding affinity were tested for antidepressant‐like behavior in wild‐type mice and animals deficient in NR2B subunit expression. In the tail suspension test, four RO‐25‐6981 analogs (TR‐2, TR‐4, TR‐5, and TR‐6) were found to exhibit antidepressant‐like activity in wild‐type mice following acute administration (30 mg/kg, i.p., 30 min) with maximal reductions in immobility by approximately half compared to vehicle‐treated controls. By contrast, RO‐25‐6981 (10 mg/kg, i.p., 30 min) reduced immobility by approximately 90%, an effect comparable to that exhibited by the traditional monoaminergic antidepressants fluoxetine and desipramine. However, unlike RO‐25‐6981, TR‐2, TR‐4, TR‐5 and TR‐6 profoundly limited generalized locomotor activity suggesting increased activity in the tail suspension test was related to psychotropic vs. generalized drug effects. In contrast, other TR analogs tested showed no antidepressant‐like activity in the tail suspension test, despite possessing robust NMDA receptor antagonist activity via mid‐ to low‐nanomolar binding affinity at the NR2B subunit. Interestingly, RO‐25‐6981 and TR‐5 exhibited similar antidepressant‐like activity in wild‐type and NR2B‐deficient mice, despite possessing low‐nanomolar NR2B binding. In addition, cellular serotonin transport assays showed functional inhibitory activity of both agents. Taken together, these data suggest that the antidepressant‐like activity of RO‐25‐6981 and its analogs does not correlate with the degree of NMDA receptor antagonism. Furthermore, these data point to monoamine reuptake inhibition contributing to the overall antidepressant‐like activity of RO‐25‐6981 in animal models of mood.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Antidepressant-Like Action of Single Facial Injection of Botulinum Neurotoxin A is Associated with Augmented 5-HT Levels and BDNF/ERK/CREB Pathways in Mouse Brain.

The present study was designed to examine the therapeutic effects of Botulinum neurotoxin A (BoNT/A) on depression-like behaviors in mice and to explore the potential mechanisms. These results revealed that a single facial injection of BoNT/A induced a rapid and prolonged improvement of depression-like behaviors in naïve and space-restriction-stressed (SRS) mice, reflected by a decreased duration of immobility in behavioral despair tests. BoNT/A significantly increased the 5-hydroxytryptamine (5-HT) levels in several brain regions, including the hippocampus and hypothalamus, in SRS mice. BoNT/A increased the expression of the N-methyl-D-aspartate receptor subunits NR1 and NR2B in the hippocampus, which were significantly decreased in SRS mice. Furthermore, BoNT/A significantly increased the expression of brain-derived neurotrophic factor (BDNF) in the hippocampus, hypothalamus, prefrontal cortex, and amygdala, which were decreased in SRS mice. Finally, BoNT/A transiently increased the levels of phosphorylated extracellular signal-regulated kinase (p-ERK) and cAMP-response element binding protein (p-CREB), which were suppressed in the hippocampus of SRS mice. Collectively, these results demonstrated that BoNT/A treatment has anti-depressant-like activity in mice, and this is associated with increased 5-HT levels and the activation of BDNF/ERK/CREB pathways in the hippocampus, supporting further investigation of BoNT/A therapy in depression.

The gap junction inhibitor INI-0602 attenuates mechanical allodynia and depression-like behaviors induced by spared nerve injury in rats.

Gap junctions (GJs) are novel molecular targets for pain therapeutics due to their pain-promoting function. INI-0602, a new GJ inhibitor, exerts a neuroprotective role, while its role in neuropathic pain is unclear. The objective was to investigate the analgesic role and mechanisms of INI-0602 in neuropathic pain induced by spared nerve injury (SNI), and whether INI-0602 attenuated pain-induced depression-like behaviors. Rats were randomly assigned to saline treatment groups (sham+NS and SNI+NS) or INI-0602 treatment groups (sham+INI-0602 and SNI+INI-0602). The von Frey test was used to assess pain behavior, and the sucrose preference test, the forced swimming test, and the tail suspension test were used to assess depression-like behaviors. Gap junction intercellular communication (GJIC) was measured by parachute assay. Western blots were used to determine the protein expression. In vitro, INI-0602 significantly suppressed GJIC by decreasing connexin43 and connexin32 expression. In vivo, INI-0602 significantly suppressed mechanical allodynia during initiation (7 days after SNI) and the maintenance phase (21 days after SNI) and simultaneously attenuated accompanying depression-like behaviors. Furthermore, INI-0602 markedly suppressed the activation of astrocytes and microglia on days 7 and 21 by reducing GJIC. Finally, INI-0602 reversed the changes in the brain-derived neurotrophic factor and Nr2b subunits of the N-methyl-D-aspartate receptor in SNI rats, suggesting that these effects of INI-0602 were related to its analgesic effect. Our findings demonstrated that blocking GJs with INI-0602 attenuated mechanical pain hypersensitivity and related depression-like behaviors in SNI rats by reducing glial activation.