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Retinal microglia behavior in mouse oxygen-induced retinopathy model:quantitation and distribution

Retinal microglia behavior in mouse oxygen-induced retinopat...

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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： Jin Liu Sookja Kim Chung Amy Cheuk Yin Lo Department of Ophthalmology Li Ka Shing Faculty of MedicineThe University of Hong Kong Department of Anatomy Li Ka Shing Faculty of MedicineThe University of Hong Kong Research Centre of Heart BrainHormone & Healthy AgingLi Ka Shing Faculty of MedicineThe University of Hong Kong State Key Laboratory of Pharmaceutical Biotechnology The University of Hong Kong

Retinopathy of Prematurity(ROP) is a principle cause of childhood blindness worldwide. Retinal microglia are immune cells in the retina, which maintains the homeostasis of local microenvironment. The association between retinal microglia and ROP has been noted but yet unclear. We performed a more detailed analysis on retinal microglia's quantitative and distributive patterns in a mouse oxygen-induced retinopathy(OIR) model, a well-established rodent model, which mimics ROP. Postnatal day(P) 7 C57BL/6J mouse pups with their nursing mother were exposed to 75% oxygen for 5 days to induce OIR. Room air(RA)-treated pups served as controls. On P12, P17, and P21, retinal microglia and vessels were visualized by immunostaining in whole-mounted retinae under a confocal microscope. Activated(amoeboid) and resting(ramified) microglia in different retinal areas(superficial central/mid-peripheral, deep central/mid-peripheral) were identified, categorized and counted. On P12, OIR retinae showed more amoeboid microglia in superficial central and mid-peripheral areas. On P17, OIR retinae showed increased amoeboid and total, but reduced ramified microglia in superficial central areas;in superficial mid-peripheral retina, OIR led to an increase in amoeboid and total microglia numbers, but a decrease in the number of ramified microglia in tufts areas;in mid-peripheral non-tufts areas, OIR led to increased amoeboid, ramified and total microglia;more ramified microglia were observed in deep OIR retinae. On P21, more microglia were observed in central and mid-peripheral superficial retina with more amoeboid form in OIR retinae. Our observations suggest that(1) retinal microglial activation continues throughout OIR;(2) activated microglia are associated with vascular abnormalities;(3) the activation of microglia persists after full regrowth of retinal vessels in OIR-challenged retina.

关键词： retinopathy of prematurity oxygen-induced retinopathy microglia

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Laminar processing of global visual contours in V1 and V2

Laminar processing of global visual contours in V1 and V2

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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： Ru-Jia Chen Feng Wang Hua-Lou Liang Wu Li State Key Laboratory of Cognitive Neuroscience and Learning and IDG/McGovern Institute for Brain ResearchBeijing Normal University School of Biomedical Engineering Drexel University

Increasing evidence indicates that both feedforward and feedback connections play important roles in contour integration, but the underlying intra- and inter-cortical circuitry is still unclear. Using a pair of laminar electrodes（U-probes, Plexon Inc）, we simultaneously recorded multiunit activities from different layers in monkey V1 and V2 at retinotopically matched locations. The monkeys were trained to detect visual contours formed by different number of collinear bars embedded in a background of randomly oriented bars. Referring to the known anatomical structures and using current source density analysis, we separated each cortical area into three laminae: supra-granular（SG）, granular（G） and infra-granular（IG） layers. Cells showing responses significantly correlated with contour length were defined as the contour cells. The distribution of contour cells across cortical layers were different. V1 contour cells were mainly located within the SG and IG layers, scarcely seen in the G layer, while in V2 the majority of contour cells were confined within the SG, G and upper IG layers. The time course and the strength of contour-related signals also varied across different layers. Contour-related signals did not emerge until 00 ms after stimulus onset for almost all V1 contour cells, and the contour signals became progressively earlier and stronger towards the upper SG. In V2 a small proportion（0%）of contour cells in the SG and G layers started to signal the global contour since their visual response onset（0 ms after stimulus onset）;the contour-related signals in the remaining cells were much delayed, similar to V1 contour cells, but the shortest latencies and maximal strengths of contour signals were seen in the lower SG layer of V2. The overall contour signals in V2 were much stronger than V1. The global contour induced a Mexican-hatresponse profile in both V1 and V2, facilitating neurons with receptive fields on the contour and suppressing those on the background. Th

关键词： laminar V1 V2 contour integration

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Presenilin1 mutants alter the cleavage pathway of longer forms of Amyloidβ protein

Presenilin1 mutants alter the cleavage pathway of longer for...

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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： Nuomin Li Kefu Liu Yunjie Qiu Zhenzhen Sun Fan Zeng Hong Qing School of Life Science Beijing Institute of Technology

Objective Alzheimer disease is characterized by progressive memory loss and damaging of cognitive functions. One of the hallmarks of AD is excessive production of Amyloid β-peptide(Aβ). Aβ is a 38～43 amino acid peptide which is derived from the β-amyloid precursor protein(APP) through sequential cleavage of β- secretases and γ-secretases. The aspartyl protease β-site APP cleaving enzyme1(BACE1) cleaves APP mainly at a unique site, whereas the γ-secretase complex cleaves the carboxy-terminal fragment at several sites, with preference for positions 40 and 42, forming the amyloid-β1-40 and Aβ1-42 peptides. The soluble forms of Aβ42 are implied to be the toxic species that causes synaptic and neuronal damage as well as impaired memory. Large numbers of pathogenic PS1 mutations have been identified which throughout the coding sequence. More that 150 Presenilin mutations are associated with familial early-onset Alzheimer disease. PS1 mutations can alter the activity of γ-secretases on cleavage of β-amyloid precursor protein, causing the elevation of Aβ production. Previous study showed that through γ-secretases, βAPP-C-terminal fragment is cleaved at the ε-site generated primarily long fragments, Aβ48 and Aβ49, and then is followed by stepwise cleavage of every three amino acid residues at the C terminus, which include Aβ48- Aβ45- Aβ42 line and Aβ49- Aβ46- Aβ43- Aβ40 line. In this report, we have performed extensive study to examine 16 different PS1 FAD mutations and their effects on the production of Aβ through APP processing. Methods Here we use Western Blot to analyze the level of CTFs, especially C99 in different PS1 mutations. And we use ELISA to analyze the Aβ42 and Aβ40 level through the different PS1 mutation cleavage. Meanwhile,we also quantitively analyzed the triple and tetrapeptide produced by two distinct lines of long-amyloid β cleavage processing with mass spectrometry method. Results(1) the western blot results show that most of most of PS1 mutations could

关键词： Familial Alzheimer's disease γ-secretases Presenilin1 Amyloid β-peptide

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The effects of carbachol on the electrophysiological properties of auditory thalamic neurons

The effects of carbachol on the electrophysiological propert...

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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： Xiao-Feng Du Ling Liu Xin Fu Hui Li Hui-Juan Jia Xin Wang Feng Luo Hubei Key Laboratory of Genetic Regulation and Integrative Biology School of Life SciencesCentral China Normal University

The medial geniculate body(MGB) including ventral(MGBv), dorsal(MGBd) and medial divisions(MGBm), is not only a main auditory center of the thalamus but also the last stage of subcortical auditory processing in the ascending pathway. Auditory learning and experience can induce plasticity in MGB neurons, and the plastic changes are proved to depend on activation of cholinergic muscarinic receptors. However, the cellular mechanism is still unclear. Using in vitro whole-cell patch clamp recordings, we studied the effects of artificially activation of muscarinic acetylcholine receptors(m ACh R) with carbachol(CCh, 10 μM) on electrophysiological properties of MGB neurons. In total, 68 MGB neurons(MGBv, 23;MGBd, 35;MGBm, 10) were obtained. According to the membrane responses upon positive current injection, bursting(26%, n = 18) and tonic(74%, n = 50) firing patterns were identified. Although most MGB neurons(MGBv, 21/23, 91%;MGBd, 23/35, 66%;MGBm, 9/10, 90%) showed tonic firing pattern, they had significant differences in resting membrane potential(RMP), latency and time constant(p<0.01, t-test). CCh(10 μM) was applied in 43 neurons to investigate the effects of artificial activation of m ACh R on electrophysiological properties. Activation of m ACh R led to excitation(13%, n = 10), inhibition(61%, n = 26) or no effect(16%, n = 7). The excitatory neurons were depolarized and showed increase in the input resistance and the time constant(p<0.05,t-test). On the contrary, the inhibitory neurons were hyperpolarized and showed decrease in the input resistance and the time constant(p<0.05, t-test). Majority of the MGBv neurons(45%, 5/11) showed excitation, while most of the MGBd(72%, 18/25) and MGBm(57%, 4/7) neurons showed inhibition. The results indicate that activation of mA Ch R tends to facilitate the MGBv, but not the MGBd and MGBm neurons. Considering the fact that MGBv belongs to lemniscal pathway and MGBm/MGBd belongs to non-lemniscal pathway, our data indicate that ch

关键词： medial geniculate body whole-cell patch clamp recordings carbachol muscarinic acetylcholine receptors

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The induction of neuronal death by up-regulated microglial cathepsin H in LPS-induced neuroinflammation

The induction of neuronal death by up-regulated microglial c...

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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： Kai Fan Daobo Li Yanli Zhang Chao Han Junjie Liang Changyi Hou Hongliang Xiao Kazuhiro Ikenaka Jianmei Ma Department of Anatomy of Dalian Medical University Clinical Medicine of Seven-year Education Dalian Medical University Regenerative Medicine Center the First Affiliated Hospital of Dalian Medical University Graduate School of Dalian Medical University Division of Neurobiology and Bioinformatics National Institute for Physiological Sciences

Background Neuroinflammation is a hallmark that leads to selective neuronal loss and/or dysfunction in neurodegenerative disorders. Microglia-derived lysosomal cathepsins are increasingly recognized as important inflammatory mediators to trigger signaling pathways in a pathological cascade that aggravate neuroinflammation in neurodegeneration. However, cathepsin H(Cat H), a cysteine protease, has been far less studied in neuroinflammation, compared to cathepsin B, D, L and S. The expression patterns and functional roles of Cat H in the brain in neuroinflammation remains unknown. Methods C57BL/6J mice were intraperitoneally injected with either 0.9% saline or lipopolysaccharide(LPS, 5 mg/kg). Immunohistochemistry(IHC) and in situ hybridization(ISH) were used to analyze regional expression and cellular localization of Cat H in the brain at different time points after LPS injection. Nitrite assay was used to examine microglial activation in vitro;ELISA was used to determine the expression and release of Cat H and proinflammatory cytokines(TNF-α, IL-1β, IL-6, IFN-γ). Cat H activity was analyzed by cellular Cat H assay kit. Flow cytometry and In situ cell death detection were used to investigate neuronal death. Results Cat H m RNA was only present in perivascular microglia and non- parenchymal sites under normal conditions. After LPS injection, Cat H m RNA expression in activated microglia in different brain regions was increased. Twenty-four hours after LPS injection, Cat H m RNA expression was maximal in SN;seventy-two hours later, it peaked in cerebral cortex and hippocampus, then decreased and maintained at a low level. The expression of Cat H protein exhibited the similar alterations at different time-points after LPS injection. In vitro, inflammatory stimulation( LPS, TNF-α, IL-1β, IL-6 and IFN-γ) increased release and activity of Cat H in microglia. Conversely, addition of Cat H to microglia promoted production and release of NO, IL-1β and IFN-γ which could be pre

关键词： cathepsin H microglia cytokines inflammation apoptosis

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Building an elevated platform for simultaneously assessing rodent locomotor activity and anxiety

Building an elevated platform for simultaneously assessing r...

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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-Feng Ding Lu Qian Hai-Hong Yang Shu-Fang Feng Yan Wu Yu-Jie Zang Ming Fan Haitao Wu Department of Neurobiology Institute of Basic Medical SciencesAcademy of Military Medical Sciences Department of Cognitive Sciences Institute of Basic Medical SciencesAcademy of Military Medical Sciences Key Laboratory of Neuroregeneration Co-innovation Center of NeuroregenerationNantong University

Elevated plus maze(EPM) task is the common behavioral assay to study rodent anxiety. However, the traditional EPM task shows some limited sensitivity. To further improve the sensitivity of EPM assay for better assessing animal anxiety, here we reported a novel elevated platform(EP) we have designed and created for better assessment of mouse anxiety. It was designed based on the principle of EPM task. Similar to EPM, the EP task utilizes the psychiatric conflict between intrinsic exploring desire and fearful challenge produced by 100 cm height to analyze the rodent anxiety preference between different group of mice. The novel elevated platform was divided into three consecutive concentric ring zones including center zone, middle zone and outer zone. The fearful challenge elevated gradually from the center zone towards the outer zone. Based on the difference of anxiety preference between female and male mice. To test our newly designed EP task, we firstly measured the spontaneous activity with open field(OF) and EP task respectively. Thereafter, the anxiety differences between female and male C57/BL6 J mice were measure by EPM and EP task analysis respectively. Our results demonstrated that EP shows significantly improved sensitivity in assessing rodent anxiety levels compared to EPM, while equivalent sensitivity to OF in assessing rodent locomotor activity. Furthermore, our newly designed EP could combine OF and EPM tasks into one for simultaneously assessing locomotion and anxiety, which will greatly simplify future rodent behavioral assay procedures. Moreover, compared to EPM task, it will be much easier to handle mouse in EP task and thus will greatly reduce animal stress duirng experiment. Together, our newly designed tool has potential powerful applications in behavioral assay of anxiety disorders and related therapeutic studies.

关键词： elevated plus maze elevated platform locomotion anxiety

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A novel full-length Htt mouse model comprehensively recapitulates Huntington's disease pathology

A novel full-length Htt mouse model comprehensively recapitu...

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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： Sushuang Zheng Sushila A Shenoy Wencheng Liu Christopher A Ross Wenzhen Duan Chenjian Li School of Life Sciences Peking University Department of Neuroscience Weill Cornell Graduate School of Medical Sciences Department of Neuroscience Johns Hopkins University

Many genetic mouse models of Huntington's disease(HD) have been created but none fully recapitulate the cardinal phenotypes of HD. Transgenic mice expressing exon 1 of huntingtin show a severe phenotype but do not replicate the specific pattern of neuropathology characteristic of HD. Models which express full-length huntingtin exhibit mild phenotypes which only represent early-stages of disease and in several cases, have non-HD phenotypes such as obesity. A full-length model with an early, robust phenotype is important for in vivo investigations into HD pathogenesis. We have created a full-length model, BAC226 Q, which has a robust HD-like phenotype including reduced life span, weight loss, motor and non-motor neurologic phenotypes, neuropathology including robust striatal neuron cell death loss. The HD motor disorder is biphasic, with early hyperkinesia and a late hypokinetic stage. BAC226 Q have normal motor function at 2 months and severe hyperkinesia including involuntary choreiform movement by 4 months. At 10 months, BAC226 Q mice are no longer hyperkinetic and show reduced activity in the open fieldcompared to non-transgenic siblings. BAC226 Q mice exhibit mood and cognitive phenotypes at 2 months, prior to motor onset. At 11 months BAC226 Q mice exhibit significant striatal cell death, the degeneration of specific interneuron's populations such as parvalbumin(PV), calbindin-D28k(CB) and calretinin(CR) positive interneurons in the striatum and cortex, widespread huntingtin inclusion pathology and reactive gliosis. At 12 months, we measure specific atrophy of the striatum and cortex, but not the cerebellum. BAC226 Q mice exhibit HD-like peripheral abnormalities such as significant body weight loss, and skeletal-muscle and myocardium atrophy. To conclude, the novel BAC226 Q mouse accurately recapitulates the full spectrum of HD phenotypes, and will be a valuable tool for studying the underlying pathogenic mechanisms of HD and for the development of HD therapies.

关键词： Huntington's disease(HD) mouse model BAC226Q

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AQP1 and EAAT2 involved in glutamate induced nuclear swelling in astrocytes

AQP1 and EAAT2 involved in glutamate induced nuclear swellin...

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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： Feng Jiang Rui Chao Chal Chun Zhang Yang Xi Hua Jia Kai GaO Albert Cheung Hoi Yu Neuroscience Research Institute Peking University Department of Neurobiology School of Basic Medical SciencesPeking University Key Laboratory for Neuroscience of the Ministry of Education and Key Laboratory for Neuroscience of the National Health and Family Planning Commission Peking University National Institute of NeurologicalDisorders and Stroke National Institutes of Health (NINDS)

Glutamate(Glu), the most important excitatory neurotransmitter in the central nervous system(CNS), mediates synaptic plasticity, learning and memory. Astrocytes remove Glu from extracellular fluid to protect neurons from toxicity. Glu uptake by astrocytes is involved in glutamine synthesis and metabolism of the TCA cycle. But lots of intracellular metabolisms of Glu and Glu-mediated neuron and astrocyte communication are unknown. In this study, we performed 3H-Glu and immunostaining experiments to demonstrate that internalized Glu in astrocytes could enter the nuclei, and directly trigger nuclear swelling and gene expression. This swelling was found to require aquaporin 1(AQP1, a water channel protein) and excitatory amino-acid transporter 2(EAAT2), and was Na+/K+ dependent. AQP1 and EAAT2 were detected on the astrocytic nuclear membrane. Over-expression of AQP1 in astrocytes enhanced Glu-induced nuclear swelling, while its inhibition or knock-down by sh RNA reduced Glu-induced nuclear swelling. Inhibition of EAAT2 by DHK, a selective non-transportable inhibitor of EAAT2, also reduced Glu-induced nuclear swelling of astrocytes. An RNA-F.I.S.H(RNA Fluorescence in situ Hybridization) experiment indicated that Glu-induced nuclear swelling modulated gene expression in astrocytes. These results suggest that AQP1 and EAAT2 localization on astrocytes nuclear membrane might uncover more potential functions of Glu in astrocytes. And these specializations we found in astrocytes might reveal a new function of neuron-astrocyte interaction mediated by Glu.

关键词： AQP1 EAAT2 glutamate astrocyte nuclear swelling

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Activation of mGluR4 reduces proliferation and promotes neural differentiation in neural stem cells derived from rat subventricular zone

Activation of mGluR4 reduces proliferation and promotes neur...

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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： Zhichao Zhang Qian Jiao Li Wang Lingyu Zhao Yumei Tian Jianshui Zhang Jianxin Liu Haixia Lu Xinlin Chen Yong Liu Institute of Neurobiology Xi’an Jiaotong University Health Science Center The Affiliated Hospital of Xi'an Medical college Xi’an Mental Health Center

Objective The subventricular zone(SVZ) functions as the largest neurogenesis site in the central nervous system(CNS) following postnatal development and into adulthood. In the neurogenic niche, these multipotent neural stem cells(NSCs) can be activated in response to the physiological and pathophysiological stimuli, whereby might participate in CNS repair and functional recovery. Recent evidence indicates that glutamate,with high extracellular concentration during developing and postnatal neurogenic niches after CNS injury, can regulate the survival, proliferation and differentiation of NSCs. These findings arouse concerns on glutamate and its receptors in regulation of NSC biology. The recent studies show that several subtypes of m Glu Rs express in the undifferentiated NSCs in neurogenic regions, where m Glu Rs may play roles in control of their survival, proliferation and differentiation. In cerebellar and neocortical neuroprecursors, m Glu R4(group III m Glu Rs) is linked to inhibiting proliferation and promoting neuronal differentiation, but little is known on how m Glu R4 affect the NSC behaviors in postnatal SVZ. In the present study, we focus on m Glu R4 in regulation of proliferation and neuronal lineage commitment of NSCs derived from rat SVZ. Furthermore, the involvement of possible cell signaling molecules will be explored. Methods(1) Brd U labeling: To study the SVZ cells proliferation, 250 g male SD rats were given intraperitoneal injections of 50 mg/kg Brd U, then rats were victimized after 4h. To investigate the SVZ cells differentiation, rats were given intraperitoneal injection of 50 mg/kg Brd U 4 times in one day before operation.(2) Stereotaxic injection: VU0155041 sodium salt dissolved in 0.9% normal saline(NS) at 2 m M of concentration;MSOP dissolved in NS at 5 m M;vehicle control rats were injected NS only. Then the drugs were stereotaxically injected into the right lateral ventricle(LV) using the following coordinate(Bregma: anteroposterior,

关键词： metabotropic glutamate receptor 4 neural stem cells proliferation differentiation subventricular zone

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Autophagy impeded deficits of spatial cognition induced by melamine

Autophagy impeded deficits of spatial cognition induced by m...

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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： Jingxuan Fu Tao Zhang College of Life Science Nankai University

Objective: It was reported that melamine was deliberately added to infant formula in order to increase protein content. Recently, several studies reported that melamine could cause neurotoxicity. Moreover, one of our previous studies suggested that autophagy could protect PC12 cells from melamine-induced cell death via inhibiting the excessive generation of ROS. Accordingly, the aim of the study is to investigate if autophagy could also impede the cognitive impairment in anaesthetized rats induced by melamine. Methods: Twenty-four three-week-old male Wistar rats were randomly assignedinto three groups, which were control group(CON, n=8), melamine group(MEL, n=8) and melamine-rapamycin group(MEL-RAP, n=8). The MEL rats were gavaged with melamine solution(30 mg/m L, dissolved in 1% carboxymethylcellulose –CMC) at a dose of 300 mg/kg·day for 28 consecutive days, while the animals in CON group were received the same dose of 1% CMC. The rats were intraperitoneally injected rapamycin 2mg/kg·d from the beginning day of gavaging melamine in the MEL-RAP group. Morris water maze(MWM) test, including training phase and spatial probe phase, was employed to evaluate the learning and memory. The long term potentiation(LTP) from Schaffer collaterals to CA1 region in the hippocampus was recorded. Results:During Morris water maze, the average path length were decreased with training in the all rats of three groups. However, it reduced more greatly in the CON group compared to the melamine group(day1, P<0.005;day2～5, P<0.001), while there was no significant difference of decreased average path length between CON group and MEL-RAP group. Both platform crossings and the quadrant dwell time were smaller in the MEL group( P < 0.001 & P < 0.01) than in the CON group, however, they were significantly increased in the MEL-RAP group( P < 0.01 & P < 0.05) compared to the MEL group. LTP test showed that field excitatory postsynaptic potentials(f EPSPs) slopes were significantly lower in the ME

关键词： Autophagy,melamine,Morris water maze,LTP,learning and memory

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