ABSTRACT
Purposeful induction of fever for healing, including the treatment of epilepsy, was used over 2000 years ago by Hippocrates. More recently, fever has been demonstrated to rescue behavioral abnormalities in children with autism. However, the mechanism of fever benefit has remained elusive due in large part to the lack of appropriate human disease models recapitulating the fever effect. Pathological mutations in the IQSEC2 gene are frequently seen in children presenting with intellectual disability, autism and epilepsy. We recently described a murine A350V IQSEC2 disease model, which recapitulates important aspects of the human A350V IQSEC2 disease phenotype and the favorable response to a prolonged and sustained rise in body core temperature in a child with the mutation. Our goal has been to use this system to understand the mechanism of fever benefit and then develop drugs that can mimic this effect and reduce IQSEC2-associated morbidity. In this study, we first demonstrate a reduction in seizures in the mouse model following brief periods of heat therapy, similar to what was observed in a child with the mutation. We then show that brief heat therapy is associated with the correction of synaptic dysfunction in neuronal cultures of A350V mice, likely mediated by Arf6-GTP.
Subject(s)
Epilepsy , Guanine Nucleotide Exchange Factors , Hyperthermia, Induced , Nerve Tissue Proteins , Seizures , Animals , Child , Humans , Mice , Epilepsy/therapy , Guanine Nucleotide Exchange Factors/genetics , Hot Temperature , Intellectual Disability/genetics , Mutation , Nerve Tissue Proteins/genetics , Receptors, AMPA/genetics , Seizures/therapyABSTRACT
Acupuncture has been used for treating drug addiction since the 1970s, but little is known about the mechanisms by which acupuncture affects drug cue-induced relapse. The transcription factor delta-FosB (ΔFosB) plays a critical role in behavior and pathology after chronic use of cocaine. ΔFosB regulates glutamate receptor signaling and dendritic spine morphology in animal models. This experimental study compared the effects of electroacupuncture (EA) at acupoints LI4 and LI11 with those of another potentially beneficial intervention, gabapentin (GBP), alone or in combination, on reinstatement of cocaine-induced conditioned place preference (CPP) and levels of ΔFosB and glutamate receptor subunit 2 (GluR2) expression in the nucleus accumbens (NAc). EA at LI4 and LI11 significantly prevented cue-induced cocaine CPP reinstatement, whereas needle insertion without electrical stimulation at these acupoints had no such effect. EA also significantly attenuated cocaine-induced increases in ΔFosB and GluR2 expression in the NAc. Unexpectedly, these effects were reversed when GBP was combined with EA. Treatment with EA at LI4 and LI11 prevented cocaine-induced increases in dendritic spine density in the NAc core and shell. Our results suggest that EA at LI4 and LI11 may prevent cocaine relapse by modulating ΔFosB and GluR2 expression, as well as dendritic spine density.
Subject(s)
Cocaine-Related Disorders/genetics , Electroacupuncture , Proto-Oncogene Proteins c-fos/genetics , Receptors, AMPA/genetics , Animals , Cocaine-Related Disorders/therapy , Gene Expression , Male , Mice, Inbred ICR , Up-RegulationABSTRACT
OBJECTIVE: To explore the influence of electroacupuncture (EA) on the expression of AMPA receptor subunit GluR1 in the rats with acute spinal cord injury (SCI) and explore the potential effect mechanism of EA in treatment of acute SCI. METHODS: A total of 80 SD rats were randomly divided into five groups, i.e. a sham-operation group, a model group, an AMPA antagonist (DNQX) group, an EA group and a DNQX+EA group, 16 rats in each group. The modified Allen's impacting method was adopted to prepare the rat model of acute SCI at T10. In the DNQX group, the intrathecal injection of 10 µL DNQX solution with a concentration of 1 nmol/µL was administered in 0.5 h after modeling success. In the EA group, EA (disperse-dense wave, 2 Hz/100 Hz in frequency, 0.5 mA in output current) was given at "Dazhui" (GV 14) and "Mingmen" (GV 4) in 0.5 h, 12 h and 24 h after modeling success for 30 min and totally 3 times. In the DNQX + EA group, the interventions in the above two groups were managed. The Basso, Beattie and Bresnahan locomotor rating score (BBB) was applied to evaluate the changes of locomotor function in the rats before modeling and in 6 h, 24 h and 48 h after modeling successively. Using the hematoxylin-eosin (HE) staining, the histopathological changes in the spinal anterior horn were observed in the spinal injured area. The immunofluorescence method was adopted to determine the number of GluR1 positive neuron of the spinal anterior horn. The Western blot method was used to determine the protein expression of GluR1 in the injured area. RESULTS: Compared to the sham-operation group in 6 h, 24 h and 48 h after modeling, the BBB scores were all significantly decreased in the model group (P<0.001) at the corresponding points. The BBB score was increased in each of intervention groups, but without statistical difference as compared with the model group (P>0.05). In the model group, it was found that the boundary between gray matter and white matter in the spinal anterior horn was blurred, the interstitial space enlarged, the neuron volume obviously shrunken, the cytoplasm decreased, the red stain deepened and some neuron nuclei fixed and shrunk. In the EA group, the morphology of the spinal anterior horn in the injured area was improved obviously, which was similar in the DNQX group and the DNQX + EA group. Compared with the sham-operation group, the GluR1 protein expression in the spinal injury area was increased (P<0.001) and the number of GluR1 positive neurons elevated (P<0.001) in the spinal anterior horn in the model group. Compared with the model group, in the EA group, the DNQX group and the DNQX + EA group, GluR1 protein expression was decreased (P<0.05, P<0.01) and the number of GluR1 positive neurons in the spinal anterior horn reduced (P<0.001). CONCLUSION: The intervention with EA at "Dazhui" and "Mingmen" promotes the repair of the injured nerve in the spinal anterior horn probably through inhibiting GluR1 expression in the spinal injured area in the rats with acute SCI.
Subject(s)
Electroacupuncture , Spinal Cord Injuries , Animals , Rats , Rats, Sprague-Dawley , Receptors, AMPA/genetics , Spinal Cord , Spinal Cord Injuries/genetics , Spinal Cord Injuries/therapyABSTRACT
OBJECTIVE@#To explore the influence of electroacupuncture (EA) on the expression of AMPA receptor subunit GluR1 in the rats with acute spinal cord injury (SCI) and explore the potential effect mechanism of EA in treatment of acute SCI.@*METHODS@#A total of 80 SD rats were randomly divided into five groups, i.e. a sham-operation group, a model group, an AMPA antagonist (DNQX) group, an EA group and a DNQX+EA group, 16 rats in each group. The modified Allen's impacting method was adopted to prepare the rat model of acute SCI at T@*RESULTS@#Compared to the sham-operation group in 6 h, 24 h and 48 h after modeling, the BBB scores were all significantly decreased in the model group (@*CONCLUSION@#The intervention with EA at "Dazhui" and "Mingmen" promotes the repair of the injured nerve in the spinal anterior horn probably through inhibiting GluR1 expression in the spinal injured area in the rats with acute SCI.
Subject(s)
Animals , Rats , Electroacupuncture , Rats, Sprague-Dawley , Receptors, AMPA/genetics , Spinal Cord , Spinal Cord Injuries/therapyABSTRACT
Tramadol, a weak agonist of mu-opioid receptors, causes seizure via several mechanisms. Preconditioning has been purposed to reduce the epileptic seizures in animal models of epilepsy. The preconditioning effect of tramadol on seizure is not studied yet. This study was designed to evaluate the preconditioning effect of ultra-low dose of tramadol on the seizures induced by tramadol at high dose. Furthermore, regarding the critical role of glutamate signaling in the pathogenesis of epilepsy, the effect of preconditioning on some glutamate signaling elements was also examined. Male Wistar rats received tramadol (2 mg/kg, i.p) or normal saline (1 mL/kg, i.p) in preconditioning and control groups, respectively. After 4 days, the challenging tramadol dose (150 mg/kg) was injected to all rats. Epileptic behaviors were recorded during 50 min. The expression of Norbin (as a regulator of metabotropic glutamate receptor 5), Calponin3 (as a regulator of excitatory synaptic markers), NR1 (NMDA receptor subunit 1) and GluR1 (AMPA receptor subunit 1) was measured in hippocampus, prefrontal cortex (PFC) and amygdala. Preconditioning decreased the number and duration of tremors and tonic-clonic seizures. Norbin, Calponin3, NR1 and GluR1 expression were decreased in hippocampus, and preconditioning had no effect on them. In contrast, it increased Norbin expression in PFC and amygdala, and attenuated NR1 and GluR1 upregulation following tramadol at high dose. These findings indicated that preconditioning by ultra-low dose of tramadol protected the animals against seizures following high dose of tramadol mediated, at least in part, by Norbin up regulation, and NR1 and GluR1 down regulation.
Subject(s)
Analgesics, Opioid/administration & dosage , Anticonvulsants/administration & dosage , Brain/drug effects , Receptors, AMPA/metabolism , Receptors, N-Methyl-D-Aspartate/metabolism , Seizures/prevention & control , Tramadol/administration & dosage , Analgesics, Opioid/toxicity , Animals , Anticonvulsants/toxicity , Brain/metabolism , Brain/physiopathology , Calcium-Binding Proteins/genetics , Calcium-Binding Proteins/metabolism , Disease Models, Animal , Dose-Response Relationship, Drug , Down-Regulation , Male , Microfilament Proteins/genetics , Microfilament Proteins/metabolism , Neuropeptides/genetics , Neuropeptides/metabolism , Rats, Wistar , Receptors, AMPA/genetics , Receptors, N-Methyl-D-Aspartate/genetics , Seizures/chemically induced , Seizures/metabolism , Seizures/physiopathology , Severity of Illness Index , Tramadol/toxicity , CalponinsABSTRACT
Previous researches have indicated that sleep plays a vital role in cognitive functions. Sleep deprivation (SD) causes learning and memory damage, which is associated with oxidative stress. This study was performed to investigate the neuroprotective effects of an extract of Abelmoschus manihot flower (EAM) against memory deficit induced by SD in mice. The SD model was evoked by multiple platform method for 5 days, successively. The learning and memory-improving effects of EAM were assessed by behavioral trials and the underlying mechanism was investigated by measuring the oxidative stress alteration. Our findings indicated that the SD-induced memory deficit and the EAM treatment improved the cognitive functions of mice in the object location recognition test and passive avoidance task. In addition, EAM effectively improved the activities of the antioxidant enzyme, decreased the content of malondialdehyde (MDA), and restored the protein expression of the brain-derived neurotrophic factor (BDNF), tyrosine kinase B (TrkB) and glutamate receptor 1 (GluR1) in brain tissues. In conclusion, EAM could improve the SD-evoked learning and memory impairments. The possible underlying mechanisms of EAM may be related to its antioxidant capacity and enhanced BDNF/TrkB/GluR1 levels in the hippocampal memory.
Subject(s)
Abelmoschus/chemistry , Memory Disorders/drug therapy , Plant Extracts/administration & dosage , Sleep Deprivation/complications , Animals , Antioxidants/administration & dosage , Antioxidants/isolation & purification , Brain-Derived Neurotrophic Factor/genetics , Brain-Derived Neurotrophic Factor/metabolism , Cognition/drug effects , Flowers/chemistry , Humans , Learning/drug effects , Male , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Memory/drug effects , Memory Disorders/etiology , Memory Disorders/psychology , Mice , Mice, Inbred ICR , Plant Extracts/isolation & purification , Protein-Tyrosine Kinases/genetics , Protein-Tyrosine Kinases/metabolism , Receptors, AMPA/genetics , Receptors, AMPA/metabolism , Sleep Deprivation/psychologyABSTRACT
BACKGROUND: Neuronal activity-induced changes in gene expression patterns are important mediators of neuronal plasticity. Many neuronal genes can be activated or inactivated in response to neuronal depolarization. Mechanisms that activate gene transcription are well established, but activity-dependent mechanisms that silence transcription are less understood. It is also not clear what is the significance of inhibiting these genes during neuronal activity. METHODS: Quantitative Real Time-PCR, western blot and immunofluorescence staining were performed to examine the expression of Senp1 and GluR1 in mouse cortical neurons. The alterations of Yy1 phosphorylation upon neuronal depolarization and the interaction of Yy1 with Brd4 were studied by protein co-immunoprecipitation. The regulators of Yy1 phosphorylation were identified by phosphatase inhibitors. Chromatin immunoprecipitation, in vitro DNA binding assay, luciferase assay and gene knockdown experiments were used to validate the roles of Yy1 and its phosphorylation as well as Brd4 in regulating Senp1 expression. RESULTS: We report that neuronal depolarization deactivates the transcription of the SUMO protease Senp1, an important component regulating synaptic transmission, scaling, and plasticity, through Yy1. In un-stimulated neurons, Senp1 transcription is activated by a Yy1-Brd4 transcription factor protein complex assembled on the Senp1 promoter. Upon membrane depolarization, however, Yy1 is dephosphorylated and the Yy1-Brd4 complex is evicted from the Senp1 promoter, reducing Senp1 transcription levels. Both Yy1 and Senp1 promote the expression of AMPA receptor subunit GluR1, a pivotal component in learning and memory. CONCLUSIONS: These results reveal an axis of Yy1/Brd4-Senp1 which regulates the expression of GluR1 during neuronal depolarization. This implicates a regulation mechanism in silencing gene expression upon neuronal activity.
Subject(s)
Cysteine Endopeptidases/genetics , Gene Expression Regulation/genetics , Neurons/physiology , Receptors, AMPA/genetics , YY1 Transcription Factor/genetics , Animals , Cysteine Endopeptidases/metabolism , Embryo, Mammalian/physiology , Mice, Inbred C57BL , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Receptors, AMPA/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , YY1 Transcription Factor/metabolismABSTRACT
STXBP1 and SCN2A gene mutations are observed in patients with epilepsies, although the circuit basis remains elusive. Here, we show that mice with haplodeficiency for these genes exhibit absence seizures with spike-and-wave discharges (SWDs) initiated by reduced cortical excitatory transmission into the striatum. Mice deficient for Stxbp1 or Scn2a in cortico-striatal but not cortico-thalamic neurons reproduce SWDs. In Stxbp1 haplodeficient mice, there is a reduction in excitatory transmission from the neocortex to striatal fast-spiking interneurons (FSIs). FSI activity transiently decreases at SWD onset, and pharmacological potentiation of AMPA receptors in the striatum but not in the thalamus suppresses SWDs. Furthermore, in wild-type mice, pharmacological inhibition of cortico-striatal FSI excitatory transmission triggers absence and convulsive seizures in a dose-dependent manner. These findings suggest that impaired cortico-striatal excitatory transmission is a plausible mechanism that triggers epilepsy in Stxbp1 and Scn2a haplodeficient mice.
Subject(s)
Corpus Striatum/metabolism , Munc18 Proteins/genetics , NAV1.2 Voltage-Gated Sodium Channel/genetics , Neocortex/metabolism , Seizures/genetics , Synaptic Transmission , Action Potentials/drug effects , Animals , Anticonvulsants/pharmacology , Corpus Striatum/drug effects , Corpus Striatum/pathology , Dioxoles/pharmacology , Electroencephalography , Epilepsy, Absence/drug therapy , Epilepsy, Absence/genetics , Epilepsy, Absence/metabolism , Epilepsy, Absence/physiopathology , Ethosuximide/pharmacology , Gene Expression Regulation , Haploinsufficiency , Interneurons/drug effects , Interneurons/metabolism , Interneurons/pathology , Mice , Mice, Knockout , Munc18 Proteins/deficiency , NAV1.2 Voltage-Gated Sodium Channel/deficiency , Neocortex/drug effects , Neocortex/pathology , Neural Pathways/drug effects , Neural Pathways/metabolism , Piperidines/pharmacology , Receptors, AMPA/genetics , Receptors, AMPA/metabolism , Seizures/metabolism , Seizures/physiopathology , Seizures/prevention & control , Signal Transduction , Thalamus/drug effects , Thalamus/metabolismABSTRACT
Centella asiatica (CA) is a widely used traditional herb, notably for its cognitive enhancing effect and potential to increase synaptogenesis. The α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) and N-methyl-D-aspartate receptors (NMDARs) mediate fast excitatory neurotransmission with key roles in long-term potentiation which is believed to be the cellular mechanism of learning and memory. Improved learning and memory can be an indication to the surface expression level of these receptors. Our previous study demonstrated that administration of CA extract improved learning and memory and enhanced expression of AMPAR GluA1 subunit while exerting no significant effects on GABAA receptors of the hippocampus in rats. Hence, to further elucidate the effects of CA, this study investigated the effects of CA extract in recognition memory and spatial memory, and its effects on AMPAR GluA1 and GluA2 subunit and NMDAR GluN2 A and GluN2B subunit expression in the entorhinal cortex (EC) and hippocampal subfields CA1 and CA3. The animals were administered with saline, 100 mg/kg, 300 mg/kg, and 600 mg/kg of CA extract through oral gavage for 14 days, followed by behavioural analysis through Open Field Test (OFT), Novel Object Recognition Task (NORT), and Morris Water Maze (MWM) and lastly morphological and immunohistochemical analysis of the surface expression of AMPAR and NMDAR subunits were performed. The results showed that 14 days of administration of 600 mg/kg of CA extract significantly improved memory assessed through NORT while 300 mg/kg of CA extract significantly improved memory of the animals assessed through MWM. Immunohistochemical analysis revealed differential modulation effects on the expressions of receptor subunits across CA1, CA3 and EC. The CA extract at the highest dose (600 mg/kg) significantly enhanced the expression of AMPAR subunit GluA1 and GluA2 in CA1, CA3 and EC, and NMDAR subunit GluN2B in CA1 and CA3 compared to control. At 300 mg/kg, CA significantly increased expression of AMPAR GluA1 in CA1 and EC, and GluA2 in CA1, CA3 and EC while 100 mg/kg of CA significantly increased expression of only AMPAR subunit GluA2 in CA3 and EC. Expression of NMDAR subunit GluN2 A was significantly reduced in the CA3 (at 100, 300, and 600 mg/kg) while no significant changes of subunit expression was observed in CA1 and EC compared to control. The results suggest that the enhanced learning and memory observed in animals administered with CA was mainly mediated through increased expression of AMPAR GluA1 and GluA2 subunits and differential expression of NMDAR GluN2 A and GluN2B subunits in the hippocampal subfields and EC. With these findings, the study revealed a new aspect of cognitive enhancing effect of CA and its therapeutic potentials through modulating receptor subunit expression.
Subject(s)
Centella , Entorhinal Cortex/metabolism , Hippocampus/metabolism , Plant Extracts/pharmacology , Receptors, AMPA/biosynthesis , Receptors, N-Methyl-D-Aspartate/biosynthesis , Spatial Memory/drug effects , Animals , Dose-Response Relationship, Drug , Entorhinal Cortex/drug effects , Gene Expression , Hippocampus/drug effects , Locomotion/drug effects , Locomotion/physiology , Male , Plant Extracts/isolation & purification , Rats , Rats, Wistar , Receptors, AMPA/genetics , Receptors, N-Methyl-D-Aspartate/genetics , Spatial Memory/physiologyABSTRACT
Orofacial pain is associated with peripheral and central sensitization of trigeminal nociceptive neurons. Nerve injury results in release of chemical mediators that contribute to persistent pain conditions. The activation of the transient receptor potential vanilloid 1 (TRPV1), promotes release of calcitonin gene-related peptide (CGRP) and substance P (SP) from trigeminal nerve terminals. CGRP and SP contribute to the development of peripheral hyperalgesia. The expression of SP and CGRP by primary afferent neurons is rapidly increased in response to peripheral inflammation. CGRP receptor activation promotes activation of AMPA receptors, leading to increased firing of neurons which is reflected as central sensitization. In this study we investigated whether inferior alveolar nerve (IAN) injury influences AMPA receptors, CGRP, SP and TRPV1 expression in the trigeminal ganglion (TG). The relative expression of the protein of interest from naive rats was compared to those from injured rats and animals that received low level laser therapy (LLLT). IAN-injury did not change expression of GluA1, GluA2 and CGRP, but increased the expression of TRPV1 and SP. LLLT increases GluA1 and GluA2 expression and decreases TVPV1, SP and CGRP. These results, together with previous behavioral data, suggest that IAN-injury induced changes in the proteins analyzed, which could impact on nociceptive threshold. These data may help to understand the molecular mechanisms of pain sensitization in the TG.
Subject(s)
Facial Nerve Injuries/radiotherapy , Gene Expression Regulation/radiation effects , Low-Level Light Therapy , Mandibular Nerve/radiation effects , Trigeminal Ganglion/radiation effects , Animals , Calcitonin Gene-Related Peptide/genetics , Calcitonin Gene-Related Peptide/metabolism , Facial Nerve Injuries/genetics , Facial Nerve Injuries/metabolism , Facial Nerve Injuries/pathology , Male , Mandibular Nerve/metabolism , Mandibular Nerve/pathology , Neurons, Afferent/metabolism , Neurons, Afferent/pathology , Neurons, Afferent/radiation effects , Photic Stimulation/methods , Rats , Rats, Sprague-Dawley , Receptors, AMPA/genetics , Receptors, AMPA/metabolism , Signal Transduction , Substance P/genetics , Substance P/metabolism , TRPV Cation Channels/genetics , TRPV Cation Channels/metabolism , Trigeminal Ganglion/injuries , Trigeminal Ganglion/metabolismABSTRACT
Experience-driven synaptic plasticity is believed to underlie adaptive behavior by rearranging the way neuronal circuits process information. We have previously discovered that O-GlcNAc transferase (OGT), an enzyme that modifies protein function by attaching ß-N-acetylglucosamine (GlcNAc) to serine and threonine residues of intracellular proteins (O-GlcNAc), regulates food intake by modulating excitatory synaptic function in neurons in the hypothalamus. However, how OGT regulates excitatory synapse function is largely unknown. Here we demonstrate that OGT is enriched in the postsynaptic density of excitatory synapses. In the postsynaptic density, O-GlcNAcylation on multiple proteins increased upon neuronal stimulation. Knockout of the OGT gene decreased the synaptic expression of the AMPA receptor GluA2 and GluA3 subunits, but not the GluA1 subunit. The number of opposed excitatory presynaptic terminals was sharply reduced upon postsynaptic knockout of OGT. There were also fewer and less mature dendritic spines on OGT knockout neurons. These data identify OGT as a molecular mechanism that regulates synapse maturity.
Subject(s)
Hypothalamus/metabolism , N-Acetylglucosaminyltransferases/metabolism , Neurons/metabolism , Synapses/metabolism , Animals , Cells, Cultured , Dendritic Spines/metabolism , Excitatory Postsynaptic Potentials/genetics , Hypothalamus/cytology , Mice, Knockout , N-Acetylglucosaminyltransferases/genetics , Neuronal Plasticity/genetics , Presynaptic Terminals/metabolism , Rats , Receptors, AMPA/genetics , Receptors, AMPA/metabolism , Synapses/genetics , Synaptic Transmission/geneticsABSTRACT
In the stargazer mouse model of absence epilepsy, altered corticothalamic excitation of reticular thalamic nucleus (RTN) neurons has been suggested to contribute to abnormal synchronicity in the corticothalamic-thalamocortical circuit, leading to spike-wave discharges, the hallmark of absence seizures. AMPA receptor expression and function are decreased in stargazer RTN, due to a mutation of AMPAR auxiliary subunit stargazin. It is unresolved and debated, however, if decreased excitation of RTN is compatible with epileptogenesis. We tested the hypothesis that relative NMDAR expression may be increased in RTN and/or thalamic synapses in stargazers using Western blot on dissected thalamic nuclei and biochemically isolated synapses, as well as immunogold cytochemistry in RTN. Expression of main NMDAR subunits was variable in stargazer RTN and relay thalamus; however, mean expression values were not statistically significantly different compared to controls. Furthermore, no systematic changes in synaptic NMDAR levels could be detected in stargazer thalamus. In contrast, AMPAR subunits were markedly decreased in both nucleus-specific and synaptic preparations. Thus, defective AMPAR trafficking in stargazer thalamus does not appear to lead to a ubiquitous compensatory increase in total and synaptic NMDAR expression, suggesting that elevated NMDAR function is not mediated by changes in protein expression in stargazer mice.
Subject(s)
Calcium Channels/genetics , Epilepsy, Absence/pathology , Receptors, N-Methyl-D-Aspartate/metabolism , Thalamus/metabolism , Animals , Calcium Channels/metabolism , Disease Models, Animal , Epilepsy, Absence/metabolism , Male , Mice , Mutagenesis , Protein Subunits/genetics , Protein Subunits/metabolism , Receptors, AMPA/genetics , Receptors, AMPA/metabolism , Receptors, N-Methyl-D-Aspartate/genetics , Synapses/metabolismABSTRACT
It has been demonstrated that stress impairs performance of skilled reaching and walking tasks in rats due to the action of glucocorticoids involved in the stress response. Skilled reaching and walking are controlled by the primary motor cortex (M1); however, it is not known whether stress-related impairments in skilled motor tasks are related to functional and/or structural alterations within the M1. We studied the effects of single and repeated injections of corticosterone (twice daily for 7 days) on spontaneous excitatory and inhibitory postsynaptic currents (sEPSCs and sIPSCs) recorded from layer II/III pyramidal neurons in ex vivo slices of the M1, prepared 2 days after the last administration of the hormone. We also measured the density of dendritic spines on pyramidal cells and the protein levels of selected subunits of AMPA, NMDA, and GABAA receptors after repeated corticosterone administration. Repeatedly administered corticosterone induced an increase in the frequency but not in the amplitude of sEPSCs, while a single administration had no effect on the recorded excitatory currents. The frequency and amplitude of sIPSCs as well as the excitability of pyramidal cells were changed neither after single nor after repeated corticosterone administration. Treatment with corticosterone for 7 days did not modify the density of dendritic spines on pyramidal neurons. Corticosterone influenced neither the protein levels of GluA1, GluA2, GluN1, GluN2A, and GluN2B subunits of glutamate receptors nor those of α1, ß2, and γ2 subunits of the GABAA receptor. The increase in sEPSCs frequency induced by repeated corticosterone administration faded out within 7 days. These data indicate that prolonged administration of exogenous corticosterone selectively and reversibly enhances glutamatergic, but not GABAergic transmission in the rat motor cortex. Our results suggest that corticosterone treatment results in an enhancement of spontaneous glutamate release from presynaptic terminals in the M1 and thereby uncovers a potential mechanism underlying stress-induced motor functions impairment.
Subject(s)
Corticosterone/pharmacology , Excitatory Postsynaptic Potentials , Glutamic Acid/metabolism , Inhibitory Postsynaptic Potentials , Motor Cortex/drug effects , gamma-Aminobutyric Acid/metabolism , Animals , Cells, Cultured , Corticosterone/administration & dosage , GABAergic Neurons/drug effects , GABAergic Neurons/metabolism , GABAergic Neurons/physiology , Male , Motor Cortex/cytology , Motor Cortex/metabolism , Pyramidal Cells/drug effects , Pyramidal Cells/metabolism , Pyramidal Cells/physiology , Rats , Rats, Wistar , Receptors, AMPA/genetics , Receptors, AMPA/metabolism , Receptors, GABA-A/genetics , Receptors, GABA-A/metabolism , Receptors, N-Methyl-D-Aspartate/genetics , Receptors, N-Methyl-D-Aspartate/metabolism , Synapses/drug effects , Synapses/metabolism , Synapses/physiologyABSTRACT
Bacopa monnieri extract has been implicated in the recovery of memory impairments due to various neurological disorders in animal models and humans. However, the precise molecular mechanism of the role of CDRI-08, a well characterized fraction of Bacopa monnieri extract, in recovery of the diabetes mellitus-induced memory impairments is not known. Here, we demonstrate that DM2 mice treated orally with lower dose of CDRI-08 (50- or 100 mg/kg BW) is able to significantly enhance spatial memory in STZ-DM2 mice and this is correlated with a significant decline in oxidative stress and up regulation of the AMPA receptor GluR2 subunit gene expression in the hippocampus. Treatment of DM2 mice with its higher dose (150 mg/kg BW or above) shows anti-diabetic effect in addition to its ability to recover the spatial memory impairment by reversing the DM2-induced elevated oxidative stress and decreased GluR2 subunit expression near to their values in normal and CDRI-08 treated control mice. Our results provide evidences towards molecular basis of the memory enhancing and anti diabetic role of the Bacopa monnieri extract in STZ-induced DM2 mice, which may have therapeutic implications.
Subject(s)
CA3 Region, Hippocampal/metabolism , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 2/metabolism , Hypoglycemic Agents/pharmacology , Plant Extracts/pharmacology , Receptors, AMPA/metabolism , Animals , Bacopa/chemistry , CA3 Region, Hippocampal/drug effects , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Experimental/psychology , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/psychology , Drug Evaluation, Preclinical , Gene Expression/drug effects , Hypoglycemic Agents/therapeutic use , Male , Malondialdehyde/metabolism , Maze Learning , Memory Disorders/drug therapy , Memory Disorders/metabolism , Mice , Oxidative Stress , Plant Extracts/therapeutic use , Receptors, AMPA/genetics , Spatial Memory/drug effects , StreptozocinABSTRACT
We investigated whether glutamate receptor subunit 2 (GluR2) is involved in EA pretreatment-induced neuroprotection via cannabinoid CB1 receptors (CB1R) after global cerebral ischemia in mice. Two hours after electric acupuncture (EA) pretreatment, global cerebral ischemia (GCI) was induced by bilateral common carotid artery occlusion (BCCAO) for 20â min. The GluR2 expression was examined in the hippocampus after reperfusion. Cell survival, neuronal apoptosis, the Bax/Bcl-2 ratio and neurological scores were evaluated at 24â h after BCCAO in the presence or absence of the GluR2 inhibitor. Furthermore, the GluR2 was determined in the presence and absence of CB1R inhibitor. Our results showed EA pretreatment enhanced expression of GluR2 in the hippocampus 2â h after reperfusion. Moreover, EA pretreatment improved neurological outcome, promoted cell survival, inhibited neuronal apoptosis, and decreased the Bax/Bcl-2 ratio after reperfusion. GluR2 knockdown by GluR2 siRNA effectively reversed the beneficial effects of EA pretreatment. Furthermore, CB1R siRNA and two CB1R antagonists blocked the elevation of GluR2 expression by EA pretreatment, whereas the two CB1R agonists up-regulated GluR2 expression as EA pretreatment. In conclusion, GluR2 up-regulation is involved in neuroprotection of EA pretreatment against GCI through CB1R, suggesting that GluR2 may be a novel target for stroke intervention.
Subject(s)
Electroacupuncture , Gene Expression Regulation , Receptor, Cannabinoid, CB1/metabolism , Receptors, AMPA/genetics , Animals , Apoptosis/drug effects , Apoptosis/genetics , Arachidonic Acids/pharmacology , Brain Ischemia/genetics , Brain Ischemia/metabolism , Brain Ischemia/therapy , Cell Survival/genetics , Disease Models, Animal , Down-Regulation , Endocannabinoids/pharmacology , Gene Knockdown Techniques , Glycerides/pharmacology , Hippocampus/metabolism , Mice , Pyramidal Cells/metabolism , RNA Interference , Receptor, Cannabinoid, CB1/agonists , Receptor, Cannabinoid, CB1/antagonists & inhibitors , Receptors, AMPA/deficiency , Reperfusion , Time Factors , Up-RegulationABSTRACT
OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on changes of spinal a-amino-3- ydroxy-5-methylisoxazole-4-propionate acid (AMPA) receptor (GluR 1) expression in rats with chronic constrictive injury (CCI) pain, so as to explore its mechanisms underlying improvement of neuropathic pain. METHODS: Sixty male SD rats were randomly divided into sham-operation, CCI model and EA groups (n=20). Neuropathic pain model was established by ligature of the right sciatic nerve. EA stimulation (2 Hz, 1-3 mA) was applied to "Weizhong" (BL 40) and "Huantiao" (GB 30) on the injured limb for 30 min, once a day for 7 days beginning from the 11th day on after CCI. The mechanical and thermal pain thresholds were measured before and after the CCI procedure (baseline) and after EA intervention. The AMPA receptor subunit GluR 1 protein and gene expression in L5-L 6 segments of the spinal cord was detected using Western blot (WB), immunohistochemistry and re- verse transcription (RT)-polymerase chain reaction (PCR), separately. RESULTS: As the results of mechanical and thermal pain thresholds in our past study, EA intervention could markedly raise CCl-reduced decreased pain threshold. Compared with the sham-operation group, the expression levels of spinal GluR 1 protein and mRNA in the model group were significantly increased (P<0.05, P<0.01). Following EA intervention, the expression levels of GluR 1 protein and mRNA were remarkably down-regulated in the EA group in comparison with those of the model group (all P<005). CONCLUSION: EA intervention can down-regulate CCI-induced increase of AMPA receptor GIuR 1 expression in the lumbar spinal cord in CCI rats, which may contribute to its effect in alleviating neuropathic pain.
Subject(s)
Electroacupuncture , Neuralgia/genetics , Neuralgia/therapy , Receptors, AMPA/genetics , Acupuncture Points , Animals , Humans , Male , Neuralgia/metabolism , Rats , Rats, Sprague-Dawley , Receptors, AMPA/metabolism , Spinal Cord/metabolism , alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid/metabolismABSTRACT
The hippocampus and the parahippocampal region have been proposed to contribute to path integration. Mice lacking GluA1-containing AMPA receptors (GluA1(-/-) mice) were previously shown to exhibit impaired hippocampal place cell selectivity. Here we investigated whether path integration performance and the activity of grid cells of the medial entorhinal cortex (MEC) are affected in these mice. We first tested GluA1(-/-) mice on a standard food-carrying homing task and found that they were impaired in processing idiothetic cues. To corroborate these findings, we developed an L-maze task that is less complex and is performed entirely in darkness, thereby reducing numerous confounding variables when testing path integration. Also in this task, the performance of GluA1(-/-) mice was impaired. Next, we performed in vivo recordings in the MEC of GluA1(-/-) mice. MEC neurons exhibited altered grid cell spatial periodicity and reduced spatial selectivity, whereas head direction tuning and speed modulation were not affected. The firing associations between pairs of neurons in GluA1(-/-) mice were stable, both in time and space, indicating that attractor states were still present despite the lack of grid periodicity. Together, these results support the hypothesis that spatial representations in the hippocampal-entorhinal network contribute to path integration.
Subject(s)
Entorhinal Cortex/cytology , Homing Behavior/physiology , Neurons/physiology , Periodicity , Receptors, AMPA/deficiency , Spatial Behavior/physiology , Acoustic Stimulation , Action Potentials/genetics , Animals , Brain Mapping , Cluster Analysis , Male , Maze Learning/physiology , Mice , Mice, Transgenic , Models, Neurological , Neural Pathways/physiology , Receptors, AMPA/genetics , Space Perception/physiology , Theta Rhythm , Time FactorsABSTRACT
A neocortical hypothesis as to homology of certain nuclear components of the avian brain proposes that the entopallium and field L2 are homologous to layer 4 of mammalian extrastriate and auditory neocortex, respectively. However, the hypothesis lacks support from the neurochemistry of thalamopallial projections. We investigated whether these projections are glutamatergic by injecting cholera toxin B into either the entopallium or field L2 in combination with in situ hybridization. Retrogradely labeled neurons in nucleus rotundus and nucleus ovoidalis were found to express vesicular glutamate transporter 2 mRNA, showing that the thalamopallial projections are glutamatergic. The results are consistent with the neocortical hypothesis.
Subject(s)
Globus Pallidus/cytology , Glutamic Acid/metabolism , Neurons/metabolism , RNA, Messenger/metabolism , Thalamus/cytology , Vesicular Glutamate Transport Protein 2/genetics , Animals , Cholera Toxin/metabolism , Columbidae , Neocortex/metabolism , Receptors, AMPA/genetics , Receptors, AMPA/metabolism , Vesicular Glutamate Transport Protein 2/metabolismABSTRACT
The aim of this study was to investigate the effect of Withania somnifera (WS) extract, withanolide A (WA), and carbamazepine (CBZ) on cerebellar AMPA receptor function in pilocarpine-induced temporal lobe epilepsy (TLE). In the present study, motor learning deficit was studied by rotarod test, grid walk test, and narrow beam test. Motor learning was significantly impaired in rats with epilepsy. The treatment with WS and WA significantly reversed the motor learning deficit in rats with epilepsy when compared with control rats. There was an increase in glutamate content and IP3 content observed in rats with epilepsy which was reversed in WS- and WA-treated rats with epilepsy. alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor dysfunction was analyzed using radiolabeled AMPA receptor binding assay, AMPA receptor mRNA expression, and immunohistochemistry using anti-AMPA receptor antibody. Our results suggest that there was a decrease in Bmax, mRNA expression, and AMPA receptor expression indicating AMPA receptor dysfunction, which is suggested to have contributed to the motor learning deficit observed in rats with epilepsy. Moreover, treatment with WS and WA resulted in physiological expression of AMPA receptors. There was also alteration in GAD and GLAST expression which supplemented the increase in extracellular glutamate. The treatment with WS and WA reversed the GAD and GLAST expression. These findings suggest that WS and WA regulate AMPA receptor function in the cerebellum of rats with TLE, which has therapeutic application in epilepsy.
Subject(s)
Anticonvulsants/therapeutic use , Epilepsy, Temporal Lobe , Learning Disabilities/etiology , Phytotherapy , Receptors, AMPA/metabolism , Withania , Withanolides/therapeutic use , Animals , Carbamazepine/therapeutic use , Cerebellum/drug effects , Cerebellum/metabolism , Disease Models, Animal , Epilepsy, Temporal Lobe/complications , Epilepsy, Temporal Lobe/pathology , Epilepsy, Temporal Lobe/therapy , Excitatory Amino Acid Transporter 1/metabolism , Gene Expression Regulation/drug effects , Glutamate Decarboxylase/metabolism , Glutamic Acid/metabolism , Inositol 1,4,5-Trisphosphate/metabolism , Linear Models , Locomotion/drug effects , Male , Motor Activity/drug effects , Motor Activity/physiology , Pilocarpine/toxicity , Protein Binding/drug effects , Psychomotor Performance/drug effects , RNA, Messenger/metabolism , Rats , Rats, Wistar , Receptors, AMPA/genetics , Time Factors , Tritium/pharmacokinetics , alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid/pharmacokineticsABSTRACT
Schizophrenia has been proposed to be associated with abnormal glutamatergic neurotransmission. The AMPA subtype of glutamate receptors (AMPARs) mediates fast excitatory synaptic transmission in the brain, and their trafficking and function is regulated in part by AMPAR auxiliary proteins including the cornichons (CNIH) and transmembrane AMPAR-regulatory proteins. Abnormal regulation of AMPARs through altered expression of these auxiliary proteins could induce changes in glutamatergic neurotransmission and thus the pathophysiology of schizophrenia. In this study, transcript expression of cornichon homologs 1-4 was measured in the dorsolateral prefrontal cortex from schizophrenia (N=25) and comparison (N=25) patient groups by comparative quantitative real-time PCR. Significant upregulation of CNIH-1, CNIH-2, and CNIH-3 mRNA expression was found in schizophrenia, with no change in CNIH-4 expression. To determine the effect of antipsychotic treatment on the expression of these genes, cornichon mRNA expression was assayed in the frontal cortex of rats treated chronically with haloperidol decanoate and no changes in any of the cornichon transcripts were found. Abnormal expression of the CNIH family of genes is consistent with cornichon-mediated AMPAR trafficking abnormalities in schizophrenia, and suggests a new mechanism contributing toward the pathophysiology of this illness.