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Downregulation of G protein coupled receptor kinase 6 mediates chronic visceral hypersensitivity through activation of adrenergic signaling pathway in rats with neonatal colonic inflammation

Downregulation of G protein coupled receptor kinase 6 mediat...

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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： Zhou Yuan Hu Shu-Fen Xu Guang-Yin Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases Institute of NeuroscienceSoochow University Department of Anesthesiology Affiliated Hospital of Nantong University

Objective The pathogenesis of irritable bowel syndrome(IBS)remains poorly understood and treatment options are very limited. The aim of present study was to explore the roles of G protein-coupled receptor kinase 6(GRK6) and β2-adrenergic receptor(β2-AR) signaling pathways in chronic visceral pain in a rat model of IBS induced by neonatal colonic inflammation(NCI). Methods Abdominal withdrawal reflex(AWR) scores to grade colorectal distention in adult male rats were recorded as a measure of visceral hypersensitivity. Western blot analysis and immunofluorescence were performed to examine protein expression. Over-expression GRK6 via lentivirus intrathecal injection was used to neutralize the effect of GRK6 in NCI model. Co-immunoprecipition assay was employed to identify the interaction between GRK6 and β2-AR. Results NCI significantly increased AWR scores and down-regulated expression of GRK6 in colonic dorsal root ganglion(DRGs) when compared with age-matched controls. NCI produced a significant elevation of norepinephrine concentration and β2-AR expression in DRGs. Administration of β2-AR antagonist butoxamine reversed NCI-induced visceral hypersensitivity. Importantly, over-expression GRK6 by intrathecal lentivirus injection markedly attenuated NCI-induced visceral hypersensitivity and reversed the up-regulation of β2-AR expression. Furthermore, co-immunoprecipitation showed the interaction between GRK6 and β2-AR. Conclusion Our data suggests that decreased expression of GRK6 mediates NCI-induced chronic visceral pain through activation of β2-AR signaling pathway. This finding may identify a potential target for treating chronic visceral pain caused by IBS.

关键词： Irritable bowel syndrome visceral hypersensitivity neonatal colonic inflammation G protein coupled receptor kinase 6 β2-adrenergic receptors dorsal root ganglion

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Neuronal representation of computation of saccadic errors in macaque lateral intraparietal cortex(LIP)

Neuronal representation of computation of saccadic errors in...

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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： Yang Zhou Yining Liu Si Wu Haidong Lu Mingsha Zhang State Key Laboratory of Cognitive Neuroscience and Learning Beijing Normal University Institute of Neuroscience Shanghai Institutes for Biological SciencesChinese Academy of Sciences University of Chinese Academy of Sciences The First Affiliated Hospital of Zhengzhou University

Objective Two types of signals in the brain code the position of motor effectors, i.e., endogenous signals(e.g., corollary discharge) and exogenous signals(e.g., proprioception). During self-oriented movements, the internally generated signals predict the required future position of motor effectors, whereas the externally generated signals inform their real positions. It has been argued that the comparison/integration of these two types of signals play a crucial role in motor control, motor learning, self-recognition and spatial perception. However, despite knowing that the frontoparietal lobes are involved in such signal integration, the underlying neural mechanisms are largely unknown. Methods: To address this question, we studied the signal integration of eye position during saccadic eye movements. We recorded single neuron's activity from the macaque lateral intraparietal cortex(LIP) while the animals were performing oculomotor tasks. LIP is a node that is used to link visual and oculomotor systems, which plays crucial roles in visual attention, saccadic planning and related cognitive processing. Results: Here, we report the co-existence of the predictive and real eye position signals in LIP: neurons with pre- and postsaccadic response predict the required future eye position;neurons with late postsaccadic response represent the extraocular proprioceptive signals. Moreover, the persistent pre- and postsaccadic response neurons behave like a "subtraction operator" to measure the discrepancy between the two types of signals. Their post-operated activity level is highly correlated with the animal's behavioral performance, e.g., the higher the post-operated activity(reflecting the greater incongruence between internal and external signals), the larger the saccadic error(distance between saccade end points and saccadic target) and the higher the probability of making corrective saccades. Conclusion: The correlation between neural activity and the animals' behavioral

关键词： Motor error Internal signal External signal Signal comparison Lateral Intraparietal Area(LIP)

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Effects of creatine on BKCa channel in primary cultured rat cortical neurons deprivated of glucose

Effects of creatine on BKCa channel in primary cultured rat ...

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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： Chen Yao Tang Guangxu Lyu Yuanyuan Cai Jiajia Zhao Li Beijing Sport University Department of Physical Education Hexi University

Objective: Large-conductance Ca2+- and voltage-activated K+(BKCa) channels are expressed in almost every tissue in central nervous system and well known for their roles in regulation of membrane excitability. The importance of BKCa channels in determination of action potential duration and frequency, and neurotransmitter release has been demonstrated partly owing to their large conductance and to the synergic activation mechanism encompassing membrane depolarization and intracellular Ca2+/Mg2+. Glycogen could be a critical energy source for neurons activity including membrane excitability. When the glucose supply is inadequate(hypoglycaemia), the resultant hypoglycaemia may cause a decrease in neuronal membrane excitability. This study observed the effects of glucose deprivation and creatine pretreatment on BKCa in primary cultured rat cortical neurons. Method: The cultured neurons were randomized into control group(C), glucose deprivation group(GD) and CGD group which was deprived of glucose after 24-hour creatine pretreatment. Whole-cell patch-clamp techniques were used to record BKCa current. The changes of intracellular Ca2+concentration([Ca2+]i) were measured by using the fluorescent imaging via laser confocal scanning microscope. Results: *** current density of BKCa significantly increased after glucose deprivation for 2h but decreased after glucose deprivation for 4h(P<0.01) compared to those of C group. While pretreated with creatine, the current density of BKCa was much lower than those in the GD group after glucose deprivation for 2-3h(P<0.01). At the same time CGD group had no significantly changes ofcurrent density with the glucose deprivation time lasting. The comparison of half activation voltage and activation rate in all groups did not show any significant differences. 2. Based on fluorescence intensity measurement, GD group exhibited significantly increased in [Ca2+]i after glucose deprivation for 1-5h(P<0.01). Creatine pretreatment significantly

关键词： primary cultured rat cortical neurons glucose deprivation creatine BKCa channel

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p38 MAPK participates in the up-regulation of GLT-1 during the induction of brain ischemic tolerance by cerebral ischemic preconditioning

p38 MAPK participates in the up-regulation of GLT-1 during t...

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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： Min Zhang Jian-Xue Gong Jia-Lei Wang Meng-Yang Jiang Li Li Hang Zhao Yu-Yan Hu Wen-Bin Li Department of Pathophysiology Hebei Medical University

Our previous study has proved that the up-regulation of glial glutamate transporter 1(GLT-1) induced by cerebral ischemic preconditioning(CIP) played an important role in the acquisition of brain ischemic tolerance in rats. However, little is known about the mechanism involved in the up-regulation of GLT-1 induced by CIP. The present study investigates whether p38 MAPK, ERK1/2, and/or JNK participates in the up-regulation of GLT-1 during the induction of brain ischemic tolerance by CIP. We found that CIP significantly enhanced the expression of p-p38 MAPK without altering p-ERK1/2 and p-JNK expression in the CA1 hippocampus. Importantly, inhibiting p38 MAPK function by SB203580, a selective inhibitor of p38 MAPK, or decreasing p38 MAPK expression by its antisense oligodeoxynucleotides(AS-ODNs) inhibited the induction of brain ischemic tolerance by CIP. The above indicated that p38 MAPK participates in the induction of brain ischemic tolerance by CIP. Furthermore, p38 MAPK was activated earlier than the up-regulation of GLT-1 in the CA1 hippocampus in rats after CIP. At the same time, the expression of p-p38 MAPK by astrocytes was increased using double immunofluoresence for p-p38 MAPK and glial fibrillary acidic protein(GFAP), a specific protein expressed by astrocytes, and p38 MAPK AS-ODNs dose dependently inhibited the up-regulation of GLT-1 in the CA1 hippocampus after CIP. Taken together, it could be concluded that p38 MAPK participates in mediating the up-regulation of GLT-1 during the induction of brain ischemic tolerance by CIP.

关键词： p38 MAPK glial glutamate transporter-1 brain ischemic tolerance cerebral ischemic preconditioning hippocampus rat

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Persistent hyperactivity of hippocampal dentate interneurons after a silent period in the rat pilocarpine model of epilepsy

Persistent hyperactivity of hippocampal dentate interneurons...

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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： Xiaochen Wang Dandan Guo Lin Wu Ren-Zhi Zhan Department of Physiology Shandong University School of Medicine

Purpose: To profile activity of GABAergic interneurons in the hippocampal dentate gyrus after the silent period following pilocarpine-induced status epilepticus(SE) in the rat. Methods: SE for more than 2 hours was induced in male Sprague-Dawley rats by intraperitoneal injection of pilocarpine. Animals were sacrificed at 1 hour, 1 week, 2 weeks or more than 10 weeks after SE to determine activity of GABAergic interneurons in the hippocampal dentate gyrus by observing immediate early gene expression and spontaneous firing at near resting membrane potential. For animals to be examined at 1 hour after SE, SE lasted for 2 hours was terminated by an intraperitoneal injection of diazepam. Key findings: c-fos and activity-regulated cytoskeletal associated protein(Arc) in hilar GABAergic interneurons were up-regulated after SE in a biphasic manner;they were increased at 1 hour and more than 10 weeks, but not 1 week after SE. Ten weeks after SE, more than 65% of hilar GABAergic cells expressed c-fos or Arc;most of them expressed c-fos and Arc more intensely than the neighboring granule cells. The percentages of calretinin(CR)-, neuronal nitric oxide synthase(n NOS)-, parvalbumin(PV)-, neuropeptide Y(NPY), and somatostatin(SOM)-positive cells expressed c-fos(compared to the control) were 19%, 27%, 28%, 49%, 51% respectively. Without the resting membrane potential to be more depolarizing in SE-induced rats, cell-attached recording mode revealed that more than 80% of hilar interneurons fired spontaneously at near resting membrane potentials(while less than 20% of the same cell population did so in controls). Significances: Pilocarpine-induced status epilepticus eventually leads to a state in whichsurviving dentate GABAergic interneurons become hyperactive with a subtype-dependent manner;this implicates a fragile balance between excitation and inhibition in the dentate gyrus and the need to strengthen GABAergic inhibition to prevent spontaneous seizures to occur. Also, for treat

关键词： GABAergic immediate early genes neuronal nitric oxide synthase neuropeptides parvalbumin spontaneous firing

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Dioxygenases Ten-eleven translocation 1(TET1) regulates oligodendrocyte differentiation

Dioxygenases Ten-eleven translocation 1(TET1) regulates olig...

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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： Jian Wang Kaixiang Zhang Ke Fang Bo Zhao Wenting Wang Shengxi Wu Xianghui Zhao Department of Neuroscience The Fourth Military Medical University

The ten-eleven translocation(TET) family of methylcytosine dioxygenases catalyze oxidation of 5-methylcytosine(5m C) to 5-hydroxymethylcytosine(5hm C) and promote DNA demethylation. Despite the abundance of 5hm C and TET proteins in the brain, little is known about their role in oligodendrocytes(OLs). We have previously shown that three TET family members possess unique subcellular and temporal expression patterns during OL differentiation;the level of 5hm C exhibits dynamic changes. Furthermore, RNAi mediated silencing of TET family proteins in OLs demonstrated that each of the TET proteins is required for OL differentiation(Glia 2014;62:914-926). Among these members, TET1 is highly expressed in oligodendrocyte precursor cells(OPCs) and early differentiated immature OL. To understand the function of TET1 in OL development, we deleted TET1 in OL lineage cells, by crossing TET1loxp/loxp with Olig1Cre+/- mice, and observed significant deficits in OL lineage formation and differentiation. In vitro cell culture indicated that loss of TET1 in OLs represses the differentiation process. In vivo myelination deficits were confirmed by electron microscopy in the optic nerve of young TET1 mutants;and the stimulated compound action potentials recorded from optic nerve were attenuated in TET1 mutants. By m RNA profiling, we observed a cohort of reduced mR NAs in the mutants. Among them, several known regulators for OL development or myelination genes were identified, such as Notch1, Opalin, Nkx2.2, Olig1, Nkx6.2, Omgp, and Tcf7l2, et al. Our study provides new insight into the regulation of OL differentiation by dioxygenases and further suggests that activation of OL differentiation regulators by TET1 is a novel mechanism for the generation of OL lineage.

关键词： Oligodendrocyte,Differentiation,Dioxygenases,TET1,Epigenetic regulation

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The coupling interface and pore domain codetermine the single-channel activity of the α7 nicotinic receptor

The coupling interface and pore domain codetermine the singl...

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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： Jun-jun Zhao Jing Qu Yuan Zheng Hui Yang Jian-liang Zhang Department of Neurobiology Beijing Institute of Brain DisordersCapital Medical UniversityKey Laboratory for Neurodegenerative Disease of the Ministry of EducationBeijing Center of Neural Regeneration and RepairBeijing Key Laboratory of Brain Major Disorders-State Key Lab Incubation BaseBeijing Neuroscience Disciplines

Objective Ligand-gated ion channels play a role in mediating fast synaptic transmission for communication between neurons. However, the structural basis for the functional coupling of the binding and pore domains, resulting in channel opening, remains a topic of intense investigation. Methods Here, a series of α7 nicotinic receptor mutants were constructed for expression in cultured mammalian cells, and their single-channel properties were examined using the patch-clamp technique combined with radio ligand binding and the fluorescence staining technique. Results We demonstrated that the replacement of the four pore-lining residues in the channel domain of the α7 nicotinic receptor with the hydrophilic residue serine prolongs the open-channel lifetime, although the conductance of these mutants decreases. At the coupling interface between the extracellular and transmembrane domains, when the VRW residues in the Cys-loop were substituted with the corresponding residues(i.e., IYN) in the 5-HT3 A receptor, thesingle-channel activity elicited by acetylcholine is impaired. This effect occurred despite the expression of the mutant receptors on the cell surface and despite the fact that the apparent Kd values were much lower than those of the wild-type α7 receptor. When we further lowered the channel-gating barrier of this chimera to enhance the open-channel probability, the loss of function was rescued. Conclusion Overall, we explored the microscopic mechanisms underlying the interplay between the channel domains and the coupling interface that affect the channel activity, and we generated an allosteric gating model for the α7 receptor. This model shows that the gating machinery and coupling assembly codetermine the single-channel gating kinetics. These results likely apply to all channels in the Cys-loop receptor family.

关键词： The coupling interface and pore domain codetermine the single-channel activity of the nicotinic receptor domain

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Abstract rule learning on rats:electrophysiological recording in A1 and ACC

Abstract rule learning on rats:electrophysiological recordin...

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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： Yan-Tian Zhang Zhi-Mei Qiao Wen Fang Li-Ping Wang Institute of Cognitive Neuroscience School of Psychology and Cognitive ScienceEast China Normal University

The ability to extract generalization rules from specific experiences is a fundamental attribute of human higher cognitive function. Humans and macaques are able to detect abstract rule changes in a sound stream. Nonprimates, it has been argued, lack such abstract rule learning ability. Previous studies have found that the primary sensory cortex is highly correlated with spatiotemporal sequence recognition and prediction. ACC is attributed to high-level cognitive ability such as anticipation of tasks, error detection and problem-solving. Here, we tested whether rats have a comparable ability by using an electrophysiological measure that has been shown to reflect higher order auditory cognition even before it becomes manifested in behavioral level. Frequently presented standard sequences had the same structure(termed XXX or XXY, where each letter represents a specific sound), and they were occasionally replaced by deviant sequences of three different types: pattern-obeying, pattern-violating and last omission sequences. On each of four training days, rats were shown 360 standard presentations of a single auditory sequence with four frequencies. On the fifth day, rats were tested by presenting 80% of the standard sequence and 20% of a kind of the deviant sequence in each block. During training and test phase, local field potentials were recorded in A1 and ACC. Each rat participated in two XXX and two XXY training-test phases. The expected results are: first, LFP in A1 and ACC shows no significant difference between standard sequences and pattern-obeying sequences;second, sequence XXY evokes MMN-like pattern in A1 and ACC;third, regardless of task(XXX and XXY), LFP patterns in both pattern-violating and last omission conditions reveal significant increase in positivity around 300 ms at ACC;finally, the last omission response in XXY task is larger than XXX task in ACC, which reveals the predictive coding on rats.

关键词： abstract rule learning A1 ACC

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LTP of EPSP evoked by ipsilateral pericentral canal stimulation in neonatal rat spinal cord motoneurons in vitro

LTP of EPSP evoked by ipsilateral pericentral canal stimulat...

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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： Lei ZHANG Huan SONG Wen QIN Yu-Han SHI Meng-Ya WANG Cell Electrophysiology Laboratory Wannan Medical College

Objective To explore the possible synaptic plasticity of excitatory synaptic transmission from ipsilateral pericentral canal(iP CC), a presumed site of "central pattern generator" of motor control, to motoneurons(MNs) in spinal cord. Methods The intracellular recordings were performed in MNs of spinal cord slices isolated from neonatal rats(8-14 days old) and the excitatory postsynaptic potentials(EPSPs) were mainly evoked by electrical stimulation of i PCC, iP CC-EPSPs. Long-term potentiation(LTP) was induced by the tetanic electrical stimulation(100 Hz, 50 pulses/train, duration 0.3～1.0 ms, 6 trains, main interval 10 seconds, 20～100 V) on i PCC. The analysis method of apparent receptor kinetics of EPSP(Acta Physiol Sin 2014, 66: 129) was used to monitor the apparent receptor kinetic properties of i PCC-EPSPs. Results In 27 MNs with i PCC-EPSPs, the tetanic electrical stimulation of i PCC enhanced the amplitude of i PCC-EPSPs to more than 120% of the baseline and the enlargement lasted longer than 30 min in 9 cells, which could be referred to as LTP(iP CC-LTP). During i PCC-LTP, apparent association rate constant(K1) increased, apparent dissociation rate constant(K2) and apparent equilibrium dissociation constant(KT) decreased only in 4 MNs. It was also observed that the induction of i PCC-LTP was facilitated by pretreatment of bicuculline(30 μM) and strychnine(1 μM), but the maintenance of i PCC-LTP was nullified by APV(30 μM and DNQX(1 μM). Conclusion These preliminary data show that the possible long-term synaptic plasticity is induced in i PCC intraspinal pathways onto MNs, which may be considered as a candidate of neural basis underlying the "central pattern generator" of motor control.

关键词： motoneuron excitatory postsynaptic potential long-term potentiation

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Sphingosine kinase 1 mediates neuroinflammation following cerebral ischemia

Sphingosine kinase 1 mediates neuroinflammation following ce...

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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： Jian Cheng Jia Jia Shuli Zheng Shanwen Wei Xiaoyu Wang Yingxiu Xu Yunqi Xiao Hui Liu Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience Soochow University College of Pharmaceutical Sciences Soochow UniversitySuzhouChina Department of Skin & Cosmetic Research Shanghai Skin Disease Hospital

Sphingosine kinases(Sphks) are the rate-limiting kinases in the generation of sphingosine-1-phosphate, which is a well-established intracellular pro-survival lipid mediator. Sphk2 has been reported to be protective following experimental stroke. We investigated the role of Sphk1 in cerebral ischemia using a mouse middle cerebral artery occlusion(MCAO) model and an in vitro glucose-oxygen deprivation(OGD) model. Sphk expression and activity were assessed in the ischemic brain with quantitative PCR(qP CR), western blot, immunohistochemistry and enzyme-linked immunosorbent assay(ELISA). Pharmacological and gene knockdown approaches were utilized to investigate the effects of Sphk1 on stroke outcomes. The expression of Sphk1 but not that of Sphk2 was rapidly induced in the cortical penumbra over 96 hours after MCAO, and the microglia were one of the major cellular sources of Sphk1 induction. Consistently, Sphk activity was enhanced in the cortical penumbra. In contrast to the protective role of Sphk2, pharmacological inhibition and cortical knockdown of Sphk1 reduced infarction at 24 and 96 hours after reperfusion. Additionally, the Sphk1 inhibitor improved the neurological deficits at 96 hours after reperfusion. Mechanistically, Sphk1 inhibition and knockdown significantly attenuated MCAO-induced expression of inflammatory mediators in the cortical penumbra. Moreover, using a conditioned medium transfer approach, we demonstrated that OGD-treated neurons induced the expression of Sphk1 and pro-inflammatory mediators in primary microglia, and the microglial induction of pro-inflammatory mediators by ischemic neurons was blunted by Sphk1 inhibition. Taken together, our results indicate that Sphk1 plays an essential role in mediating post-stroke neuroinflammation.

关键词： cerebral ischemia sphingosine kinase 1 neuroinflammation microglia

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