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Neutrophil depletion reduces neuroinflammation and attenuates delayed neuronal death and behavioral deficits after cerebral ischemia and reperfusion

Neutrophil depletion reduces neuroinflammation and attenuate...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Qun Wang Yi-Long Zhao Fang Su Wei-Wei Li An-Chen Guo Yong-Jun Wang Department of Neurology Beijing Tiantan HospitalCapital Medical University Beijing Key Laboratory of Translational Medicine for Cardiovascular Diseases Beijing Institute for Brain Disorder

Objective Neuroinflammation after cerebral ischemia/reperfusion(I/R) is characterized by the accumulation of inflammatory cells and mediators in brain. After I/R, circulating leukocytes adhere to vessel walls, leading to emigration and accumulation into ischemic brain tissue. The formation of reactive nitrogen and oxygen species and release of hydrolytic enzymes by emigrated leukocytes may contribute to secondary brain injury. Since neutrophils are the first leukocyte subtype recruited to the ischemic brain, we hypothesized that depletion of circulating granulocytes by treatment with anti-neutrophil serum(ANS) would attenuate neutrophil infiltration and glial activation, ameliorate neuronal apoptosis, and reduce behavioral deficits after cerebral I/R. Methods C57BL6 mice were administered anti-mouse ANS(5μl;ip) 24 and 2 hrs prior to I/R. Both common carotid arteries(CCA) were then occluded for 20 min followed by reperfusion for varying time periods. Circulating leukocyte counts were determined 2 hrs after the second treatment withanti-PMN serum. Hippocampal neutrophil infiltration was quantified by measuring myeloperoxidase(MPO) activity and CD18 expression(western blot) at 24 hrs after I/R. Locomotor activity was evaluated as a behavioral marker at 48 hrs after surgery while neuronal degeneration and apoptosis, astrocyte and microglial activation, and the extent of delayed neuronal death(DND) in hippocampal CA1 region were assessed 4 days after I/R. Results Circulating leukocyte counts were markedly reduced by ANS administration. Cerebral I/R induced marked neutrophil infiltration, neuronal degeneration and apoptosis, astrocyte and microglial activation, and DND in the CA1 region, and a marked behavioral deficit, effects that were significantly reduced by neutrophil depletion with anti-PMN serum. Conclusion Our results support the concept that neutrophil infiltration plays a critical role in I/R-induced neuroinflammation and secondary brain damage.

关键词： neutrophil infiltration neuroinflammation brain injury cerebral ischemia/reperfusion

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Augmentation of necroptosis dependent enzymatic activity induces spatial short term memory impairment in rats:A time-dependent study

Augmentation of necroptosis dependent enzymatic activity ind...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Sara Nikseresht Fariba Khodagholi Leila Dargahi Neuroscience Research Center Shahid Beheshti University of Medical Sciences Neuro Biology Research Center Shahid Beheshti University of Medical Sciences

Necroptosis is a novel cell death pathway and Glutamate-ammonia ligase(GLUL) and glutamate dehydrogenase-1(GLUD1) enzymes have vital functions in this type of cell death. Necroptosis is also involved in learning and memory functions and cognitive performance. We evaluated alteration of GLUL and GLUD1 activity along with spatial short-term memory impairment in rat hippocampus following LPS ***, LPS was injected in rat brain(*** 055:B55, 15μg/3μl sterile NS, i.c.v) and animals were sacrificed at different times including 4, 8, 12, 24, 48, 72 and 96 h following LPS injection;then hippocampi were removed and the levels of GLUL and GLUD1 enzymatic activity were measured by using a spectrophotometric assay. Second, to determine the relationship between spatial short term memory and necroptotic enzymes, animals were assessed by Y-maze spontaneous alternation to evaluate spatial short term memory 24 and 72 h after LPS injection. Our results showed that LPS increased GLUL activity time-dependently from 4 to 24 h continuously, decrease started in 48 h and continued in 72 h. Finally GLUL returned to the basal level in 96 h. In this way, GLUD1 enzyme activity augmented 1.8 folds in 24 h compared to its basal levels. In returning to basal level, GLUD1 followed a different time-dependent pattern and arrived to basal level in 72 h. Behavioral test results of Y-maze at 1st and 3rd day after LPS injection showed that LPS decreased spontaneous alternation by 28% compared to sham group in 3rd day while there was no significant difference between spontaneous alternation in 1st day and sham *** conclusion, there is a direct connection between necroptosis and memory performance. Following GLUL and GLUD1 activity augmentation, we observed decrease in spontaneous alternation in Y-maze test.

关键词： necroptosis hippocampus spatial memory

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De novo GRIN2A mutation results in early-onset epileptic encephalopathy:from molecular mechanism to personalized therapy

De novo GRIN2A mutation results in early-onset epileptic enc...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Hongjie Yuan Sharon A.Swanger Stephen F.Traynelis Department of Pharmacology Emory University School of Medicine

Objective N-methyl-D-aspartate(NMDA) receptors play important role in neurological diseases, including epilepsy. We report a child with an early-onset epileptic encephalopathy due to a novel de novo GRIN2 A missense mutation(p.L812M). Methods Site-directed mutagenesis, preparation of c RNA, and two-electrode voltage-clamp recordings from Xenopus oocytes were performed. The response time course was assessed by whole cell voltage-clamp current recordings on transfected HEK cells by a rapid solution exchange system. Results GluN 2A-L812M-containing receptors showed an 8-2+inhibition. The response time course(tau deactivation rate) for the mutant was 443 ms, which is 9-fold longer than 47 ms of WT. The mutant showed significantly more channel openings(open probability, 0.64 vs. 0.23 of WT). Quantification of co-transfected luciferase suggested L812 M increased neuronal death. Since conventional AEDs did not provide adequate control of the patient's seizures, a number of FDA-approved NMDAR antagonists were evaluated. Memantine inhibited the mutant and WT receptors with a similar potency, indicating that it can effectively reduce L812 M induced NMDAR hyperactivity. Based on these findings, memantine was added as an adjunct medication to the AED regimen. Memantine reduced seizure frequency with EEG improvement. Conclusion The data suggest that L812M-containing NMDARs were overactive due to the increased activation at low concentrations of agonists, enhanced currents resulted from reduced Mg2+ blockade, prolonged synapse response time, and increased channel open probability. The resulting enhanced excitatory drive is consistent with the patient's phenotype. These results indicate future evaluations of children with intractable seizures and early-onset epileptic encephalopathies should include NMDAR genes. These data also raise the possibility of using alternative AEDs(e.g. memantine) for treatment of refractory epilepsy.

关键词： NMDA receptor epilepsy channelopathy

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Rolipram facilitates long-term memory extinction and renewal but impairs working memory in water maze

Rolipram facilitates long-term memory extinction and renewal...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Xue-Fei Hou Xin Li Zhi-Yong He Wei-Yan Hu Ming Zhang The Key Laboratory of Stem Cell and Regenerative Medicine Institute of Molecular and Clinical MedicineKunming Medical University

Memory flexibility usually refers to the ability to refresh or renew acquired long-term memory. For instance, memory flexibility is required for rats or mice to find and remember the reversal platform location of the Morris water maze task(WMT). Different from memory flexibility, working memory refers to the continued refresh or renewal of short-term memory. It is supposed that the established long-term memory for the location of the initial platform would be suppressed or weakened during the reversal platform training, but not scratched as in working memory. We previously found that the overexpression of type 1 adenylyl cyclase in the forebrain of mice obviously improved the performance in locating the reversal platform of the WMT. However, it remains unclear whether the enhanced memory flexibility induced by the upregulation of intracellular cA MP-PKA signaling is purely via the improved memory acquisition, the enhanced ability to suppress consolidated long-term memory, or both of them. For better understanding of the role of c AMP signaling in the regulation of memory flexibility, we used the method of behavioral pharmacology by employing rolipram, a type IV-specific phosphodiesterase inhibitor, to selectively enhance c AMP signaling at different training stages of the WMT. Rolipram(0.1 μM/kg;i.p.) or saline was injected 1 h before behavioral training in all experiments of the present study. In Experiment 1, mice were trained in the WMT for 7 days with 1 trial per day. Rolipram treatment did not significantly improve the learning curve in locating the hidden platform, but slightly improved the performance of the probe test at day 8. In Experiment 2, all mice were first trained in the WMT for 7 days with 4 trials per day. Then, rolipram or saline was administered, and the reversal platform was introduced with 1 trial per day. Rolipram-treated mice showed the improvement in locating the reversal platform and in the probe test, supporting our previous finding that t

关键词： memory flexibility Rolipram c AMP working memory

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Optogenetic control of thalamus as a tool for interrupting penicillin induced seizures

Optogenetic control of thalamus as a tool for interrupting p...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Yechao Han Feiqiang Ma Yueming Wang Kedi Xu Qiushi Academy for Advanced Studies (QAAS) Zhejiang University Department of Emergency Second Affiliated HospitalSchool of MedicineZhejiang University

Objective Penicillin epilepsy model, whose discharge resembles that of human absence epilepsy, is one of the most useful acute experimental epilepsy models. Though closed-loop optogenetic strategy of interrupting seizures was proved sufficient to switch off epilepsy by controlling thalamus from the post-lesion partial chronic epilepsy model, doubts still exist in absence epilepsy attenuation through silencing thalamus. Here we reported direct arrest thalamus was to modulate penicillin-induced generalized epilepsy through pseudorandom responsive stimulation in e Np HR-transfected rats. Methods(1)Virus Injection(SD rats, 5-6 weeks old, r AAV-Cam KIIα-e Np HR3.0-e YFP, 1ul, 2.5×109copies, AP=-2.8mm, ML=-3.0mm, DV=-6.3mm)(2)Operation(Optrode and four transcranial screws electrodes, cannula, reference and ground channel were implanted)(3)Light Illumination(wavelength 589 nm, average power=23±3mw, in penicillin-free rats)(4)Penicillin Injection and Responsive Laser Stimulation(400UI in 2.0ul saline, 0.2ul/min, AP=-3.1mm, ML=-5.0mm,-2.3mm ventral to the skull face)(5)Perfusion and Slicing(4% paraformaldehyde, 50 um Coronal brain sections). Results(1) In penicillin-free rats, spike firing rate, amplitude of MUA and high-gamma power band of thalamic electrical signals decreased explicitly during the laser light illumination.(2) After seizures induction, the peak frequency band of intracranial EEGs in penicillin induced seizures was 9-11 Hz.(3) No effect on the time variance was witnessed, through selectively illuminating the ventrobasal thalamus, with respect to seizure duration increase or decrease between light and no light conditions. Conclusion Our data suggested that under seizure-free circumstances effective intervention of thalamus activity was observed in the variance of the spike rate, MUA and LFP by optogenetic manipulations. However, no sufficient thalamic inhibition was found in penicillin induced absence epilepsy.

关键词： optogenetics epilepsy penicillin

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Sulbactam protects hippocampal neurons against oxygen-glucose deprivation by up-regulation of GLT-1 via P38 MAPK signal pathway in astrocytes

Sulbactam protects hippocampal neurons against oxygen-glucos...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Jie Qi Min Zhang Jing-Ge Zhang Yan-Chao Liu Wen-Bin Li Department of Pathophysiology Hebei Medical University Aging and Cognition Neuroscience Laboratory of Hebei Province

Glial glutamate transporter-1(GLT-1) plays an important role in maintaining normal levels of extracellular glutamate concentration by uptake of the glutamate physiologically released. The dysfunction of GLT-1 will lead accumulation of extracellular glutamate and induce excitotoxity, which takes part in the neuronal injury after cerebral ischemia. Our recent study has shown that sulbactam, a non-typical β-lactam antibiotic, could prevent cerebral ischemic injury by up-regulating GLT-1 in a rat global cerebral ischemic model. However, the mechanisms involved in the upregulation of GLT-1 in the process are not well understood. Our previous study suggested that the upregulation of GLT-1 by cerebral ischemic preconditioning was partly due to the p38 MAPK pathway activation in rats. Astrocytes are in principle the main cell target for GLT-1 expression. Therefore, the present study was undertaken to investigate the role of p38 MAPK in the upregulation of GLT-1 during the neuronal protection against ischemic injury induced by sulbactam by neuron-astrocyte co-culture and single astrocyte culture of neonatal rats. The time of oxygen-glucose deprivation(OGD) of primary co-culture of hippocampal neurons and astrocytes and single astrocyte culture was 2h, and the profiles of neuronal death and survival were observed at 24 h after the OGD by hoechst/propidium iodide double fluorescence staining and MTT methods, respectively. The sulbactam incubation for the co-culture of neuron-astrocytes and single astrocyte culture pretreatment was persisted for48h, and then the medium was replaced by the basal medium which is free of sulbactam. In sulbactam+OGD group, the OGD was immediately induced after the sulbactam incubation for 48 h. The GLT-1 and p-p38 MAPK expressions were detected by western blot and immunocytochemistry assays in astrocyte culture in both normal and OGD states. Hoechst/propidium iodide double fluorescence staining showed that OGD induced significant neuronal death at

关键词： sulbactam neuron-astrocyte co-culture astrocyte culture oxygen glucose deprivation GLT-1 rat

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The red nucleus IL-6 in the development of neuropathic pain induced by spared nerve injury

The red nucleus IL-6 in the development of neuropathic pain ...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Cui-ping DING Jing YU Xiao-Yan ZENG Jun-Yang WANG Department of Immunology and Pathogenic Biology Xi'an Jiaotong University Health Science Center Department of Laboratory Medicine The First Affiliated HospitalXi'an Jiaotong University Health Science Center

Objective The red nucleus(RN) is an important nucleus of extracorticospinal tract, which comprises a critical subcortical relay station for a massive descending motor tract(the rubrospinal tract). Recent studies demonstrate that the RN is also involved in the modulation of neuropathic pain and plays both a facilitated role by pro-inflammatory cytokines, such as tumor necrosis factor-alpha(TNF-α) and interleukin-β(IL-β), and an inhibitory role through the anti-inflammatory cytokines including IL-10 and transforming growth factor-beta(TGF-β). Previous studies indicated that IL-6, a pro-inflammatory cytokine, plays an important role in the development of neuropathic pain induced by peripheral nerve injury. The current work was desired to investigate the expressions and roles of IL-6, as well as its receptor alpha(IL-6Rα) in the RN of rats with spared nerve injury(SNI). Methods The neuropathic pain was induced by tightly ligating the tibial and common peroneal branches of right sciatic nerve of rats. The expressions of IL-6 and IL-6R? were confirmed using immunohistochemistry, and the mean optical density and area ratio of immunoreactivity(IR) were analyzed with Image J software. To further study the roles of the RN IL-6 in the development of neuropathic allodynia induced by SNI, different doses of recombinant rat IL-6(10, 20, 40 ng) and anti-IL-6 antibody(100, 250, 500 ng) were respectively microinjected into the left RN of normal and SNI rats, and the mechanical paw withdrawal threshold(PWT) of right hind paw were dynamically assessed using von Frey filaments. Results(1) Immunohistochemistry demonstrated that both IL-6 and IL-6Rα were constitutively expressed in the RN of normal rats. At three weeks after SNI, IL-6 and IL-6Rαwere markedly up-regulated in the RN contralateral to the nerve injury side of rats. Quantitative analysis of IR showed that the mean optical density and area ratio of IL-6- and IL-6Rα-positive cells were significantly increased in the RN at three

关键词： Red nucleus Interleukin-6 Neuropathic pain Spared nerve injury

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Protective effects and mechanisms of rosmarinic acid against rotenone-induced Parkinson's disease models

Protective effects and mechanisms of rosmarinic acid against...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Wenting Jia Huamin Xu Hong Jiang JunXia Xie Department of Physiology Shandong Provincial Key Laboratory of Pathogenesis and Prevention of Neurological Disorders and State Key Disciplines: PhysiologyMedical College of Qingdao University

Parkinson's disease(PD) is a common neurodegenerative disease characterized by the loss of dopaminergic neurons in the substantia nigra(SN) and the appearance of fibrillar aggregates of α-synuclein called Lewy bodies(LBs). Rosmarinic acid(RA) is an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid, which is commonly found in various plant families. It has been demonstrated that RA has a number of interesting biological activities, e.g. antiviral, antibacterial, anti-inflammatory and antioxidant activities. However, the effect of RA on α-synuclein aggregation in PD models and the molecular mechanisms underlying this effect are still not elucidated. Therefore, in this study, we investigated the effects of RA against rotenone-induced α-synuclein aggregation in C57BL/6 mice and SH-SY5 Y dopaminergic cells and to explore the possible mechanisms involved in this process. Results showed that mice which had orally received rotenone for 56 d showed a significantly motor dysfunction;50 mg/kg RA pretreatment significantly improved the motor dysfunction. The number of TH-positive neurons and the expression of TH were both reduced in the SN of rotenone-treated mice. This could be significantly inhibited by RA pretreatment. In addition, RA pretreatment significantly inhibited rotenone-induced increase in α-synuclein expression. In vitro study showed that RA pretreatment partially restored cell loss induced by rotenone and inhibited rotenone-induced increase in α-synuclein m RNA expression in SH-SY5 Y cells. The mechanisms underlying this protection might be related to the antioxidation effect of RA by inhibiting ROS generation and increasing SODexpression, as well as promoting the degradation of alpha-synuclein by up-regulating the expression of HO-1 and HSP70. The up-regulation of the HO-1 may be connected with Nrf2 and HIF-1α. Intracellular signaling pathway MEK/ERK might participate in these processes. These results provide new findings and new strategies for the preven

关键词： Parkinson's disease α-synuclein rosmarinic acid

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Stronger within-“lazy eye” monocular inhibitory interactions in V1 of anisometropic amblyopes model contrast sensitivity losses and stronger tilt illusion

Stronger within-“lazy eye” monocular inhibitory interactio...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Tzvetomir Tzvetanov Jin-Feng Huang Cai-Yuan Liu Zhong-Jian Liu Yi-Feng Zhou CAS Key Laboratory of Brain Function and Disease & School of Life Sciences University of Science and Technology of China Research and Treatment Center of Amblyopia and Strabismus University of Science and Technology of China

Anisometropic amblyopia is a visual neural deficit that deteriorates early feature processing, object perception and binocular vision. Its neural origins begin at V1 area, hypothesized to explain contrast sensitivity function(CSF) losses of amblyopic eyes(AEs). In a companion experimental study, we found that the amount of monocular tilt illusion is strongly correlated to the monocular CSF amplitude in AEs while there was no effect in non-AEs. Since the tilt illusion is explained with V1 inhibitory interactions, we asked whether we can mechanistically explain both tilt illusion and CSF changes due to V1 neural network monocular interaction changes compared to normal eyes. First, based on physiological data of V1 relating spatial frequency and contrast sensitivity, we modelled the behavioural CSF by decoding V1 neural responses tuned to different spatial frequencies(SF, log-Gaussian function F(SF)) and contrast(c, Naka-Rushton function G(c), semi-saturation constant c50): r = r0+rmax*F(SF)*G(c)(r0, rmax are background and amplitude firing rates). Two model parameters, rmax and c50, have different effects onto the amplitude and spread of the CSF. By fitting the psychophysical data, we found that CSF amplitude of AEs is systematically explained by changes in c50 while there was no such relationship for non-AEs, that is, in AEs lower CSF amplitudes were explained through higher c50 and vice versa. Second, we investigated the network mechanism of inhibitory interactions effectsonto perceptual outcome. We found that stronger lateral inhibition between neurons increased the semi-saturation constant, i.e. decreased CSF, and also provided stronger tilt misperception effects. Overall, our computational modelling shows that stronger monocular inhibitory interactions in V1 are a sufficient explanation of the reported low-level visual deficits.

关键词： amblyopia V1 model inhibition tilt illusion contrast sensitivity

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Decoding from ECo G signals reveals the contents of color imagery in macaque inferior temporal and prefrontal cortices

Decoding from ECo G signals reveals the contents of color im...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Hisashi Tanigawa Ren Takei Kei Majima Keisuke Kawasaki Takafumi Suzuki Kiyoshi Nakahara Yukiyasu Kamitani Isao Hasegawa Cent for Transdiscipl Res Niigata Univ Grad Sch of Sci & Tech Niigata Univ ATR Grad Sch of Info Sci NAIST Niigata Univ Grad Sch of Med & Dent Sci Natl Inst of Info & Comm Tech Kochi Univ of Tech

Reading out mental content from brain activity is one of challenging goals in neuroscience. For this purpose, multichannel subdural electrocorticography(ECo G) is a promising tool because of its ability to record surface field potential with broad spatial coverage, high temporal resolution, and high signal fidelity. In this study, we are attempting to decode recalled color imagery using ECoG signals in macaques. We implanted 128- and 64-channel ECo G electrode arrays with 2.5-mm spaced grid configuration on the inferior temporal and prefrontal cortices, respectively, which are known to be important for memory recall. Monkeys performed color recall tasks. To distinguish between decoding reflecting cue information that was used for color recall and decoding reflecting recalled color imagery, we used two different cues for each color. In the task, a cue stimulus, either an achromatic scene image or a Fourier descriptor, was presented, and after a delay period, 1–2 colored shapes were presented sequentially as choice stimuli. To receive a reward, the monkey released a lever upon presentation of a correctly colored choice that was associated with the cue. The monkey was supposed to recall and maintain color imagery during the delay period. Using a power spectrum of ECo G signals during that period, we constructed support vector machine classifiers(decoders) to predict the monkey's color choice. To examine the generalization performance of the decoders across the tasks using different cues, we trained the decoders on the data set from the task using one type of cues(scene images) and tested them on the data set from the task using another type of cues(Fourier descriptors), and vice versa. Our results demonstrate that the decoders successfully predicted the monkey's color choice regardless of the cue types, providing support that recalled color imagery can be decoded by our methods. Interestingly, theta power in the prefrontal cortex, not in the inferior temporal cortex, c

关键词： Brain-machine interface,memory retrieval

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