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whole-cell recording of the robust nucleus of the arcopallium neurons in the adult zebra finch

Neural Regeneration Research 2009年第8期4卷 623-628页

作者： Suqun Liao Xiaolin Liu Dongfeng Li College of Life Sciences South China Normal University Guangzhou 510631 Guangdong Province China

BACKGROUND： Electrophysiological properties of the song nucleus have been revealed using conventional techniques, such as intracellular and extracellular recording. Research concerning the neuronal activation properties and regulations of the song system at the cellular and ion channel level may help reveal the neural mechanism of song learning. OBJECTIVE： To perform whole-cell recording of robust nucleus of the arcopallium （RA） neurons in brain slices from adult zebra finches （Taeniopygia guttata） and observe the action potential, sodium/potassium current and the spontaneous postsynaptic current of RA neurons. DESIGN, TIME AND SETTING： Self-controlled, neuroelectrophysiological experiment. The study was performed at the Neurophysiology Laboratory of South China Normal University from April to September 2008. MATERIALS： Flaming/Brown puller P-97 was purchased from Sutter Ins, USA; Axopatch 700B amplifier and Digidata 1332A converter were purchased from Axon Instrument, USA; pClamp software was provided by Axon Instrument, USA. METHODS： RA neurons were acutely isolated from 24 healthy male zebra finches. The action potential, voltage-gate sodium/potassium current and spontaneous postsynaptic current were recorded by whole-cell recording technology. Data were analyzed by pClamp software. MAIN OUTCOME MEASURES： The amplitude and frequency of the action potential, and the amplitude of the voltage-dependent and spontaneous postsynaptic currents, were measured. RESULTS：（1） Testing of action potential： cells exhibited a stable current-voltage relationship following a series of hyperpolarization stepped currents, and an action potential was triggered by the spike threshold. All the recorded cells displayed repetitive firing following depolarizing current injection, with a frequency beyond 100 Hz. （2） Testing of voltage-gate currents： The inward and outward whole-cell currents were observed after a series of depolarizing voltage steps. The inward current disappeared following the application of tetrodotoxin and the outwa

关键词： adult zebra finch robust nucleus of the arcopallium whole-cell recording

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Binocular form deprivation influences the visual cortex

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Neural Regeneration Research 2012年第34期7卷 2713-2718页

作者： Mingming Liu Chuanhuang Weng Hanping Xie Wei Qin Southwest Hospital/Southwest Eye Hospital Third Military Medical University of Chinese PLA

a-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors are considered to play a crucial role in synaptic plasticity in the developing visual cortex. In this study, we established a rat model of binocular form deprivation by suturing the rat binocular eyelids before eye-opening at postnatal day 14. During development, the decay time of excitatory postsynaptic currents mediated by a-amino-3- hydroxy-5-methyl-4-isoxazolepropionic acid receptors of normal rats became longer after eye- opening; however, the decay time did not change significantly in binocular form deprivation rats. The peak value in the normal group became gradually larger with age, but there was no significant change in the binocular form deprivation group. These findings indicate that binocular form deprivation influences the properties of excitatory postsynaptic currents mediated by a-amino-3- hydroxy-5-methyl-4-isoxazolepropionic acid receptors in the rat visual cortex around the end of the critical period, indicating that form stimulation is associated with the experience-dependent modification of neuronal synapses in the visual cortex.

关键词： a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors excitatory postsynaptic currents whole-cell recording visual cortex binocular form deprivation visual development neuraldevelopment regeneration neural regeneration

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Growth Differentiation Factor-15 Produces Analgesia by Inhibiting Tetrodotoxin-Resistant Nav1.8 Sodium Channel Activity in Rat Primary Sensory Neurons

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Neuroscience Bulletin 2021年第9期37卷 1289-1302页

作者： Wei Lin Wen-Wen Zhang Ning Lyu Hong Cao Wen-Dong Xu Yu-Qiu Zhang Department of Translational Neuroscience Jing’an District Centre Hospital of ShanghaiState Key Laboratory of Medical Neurobiology and Institutes of Brain ScienceFudan UniversityShanghai 200032China Department of Hand Surgery Huashan HospitalFudan UniversityShanghai 200040China Department of Neurobiology School of Basic Medical SciencesShanghai Medical CollegeFudan UniversityShanghai 200032China

Growth differentiation factor 15(GDF-15)is a member of the transforming growth factor-βsuperfamily.It is widely distributed in the central and peripheral nervous systems.Whether and how GDF-15 modulates nociceptive signaling remains unclear.Behaviorally,we found that peripheral GDF-15 significantly elevated nociceptive response thresholds to mechanical and thermal stimuli in naïve and arthritic rats.Electrophysiologically,we demonstrated that GDF-15 decreased the excitability of small-diameter dorsal root ganglia(DRG)neurons.Furthermore,GDF-15 concentration-dependently suppressed tetrodotoxin-resistant sodium channel Nav1.8 currents,and shifted the steady-state inactivation curves of Nav1.8 in a hyperpolarizing direction.GDF-15 also reduced window currents and slowed down the recovery rate of Nav1.8 channels,suggesting that GDF-15 accelerated inactivation and slowed recovery of the channel.Immunohistochemistry results showed that activin receptor-like kinase-2(ALK2)was widely expressed in DRG medium-and small-diameter neurons,and some of them were Nav1.8-positive.Blockade of ALK2 prevented the GDF-15-induced inhibition of Nav1.8 currents and nociceptive behaviors.Inhibition of PKA and ERK,but not PKC,blocked the inhibitory effect of GDF-15 on Nav1.8 currents.These results suggest a functional link between GDF-15 and Nav1.8 in DRG neurons via ALK2 receptors and PKA associated with MEK/ERK,which mediate the peripheral analgesia of GDF-15.

关键词： Growth differentiation factor-15 Tetrodotoxin-resistant sodium channel Nav1.8 Dorsal root ganglion whole-cell recording Activin receptor-like kinase-2 Pain

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Acetylcholine modulates transient outward potassium channel in acutely isolated cerebral cortical neurons of rats

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Neural Regeneration Research 2009年第6期4卷 436-442页

作者： Lanwei Cui Tao Sun Lihui Qu Yurong Li Haixia Wen Department of Paediatrics First Affiliated Hospital of Harbin Medical University Harbin 150001 Heilongjiang Province China Harbin Institute of Technology Harbin 150001 Heilongjiang Province China

BACKGROUND： The neuronal transient outward potassium channel has been shown to be highly associated with acetylcholine. However, the influence of acetylcholine on the transient outward potassium current in cerebral cortical neurons remains poorly understood. OBJECTIVE： To investigate acetylcholine modulation on transient outward potassium current in rat parietal cortical neurons using the whole-cell patch-clamp technique. DESIGN, TIME AND SETTING： A neuroelectrophysiology study was performed at the Department of Physiology, Harbin Medical University between January 2005 and January 2006. MATERIALS： Wistar rats were provided by the Animal Research Center, the Second Hospital of Harbin Medical University; PC-IIC patch-clamp amplifier and IBBClamp data collection analysis system were provided by Huazhong University for Science and Technology, Wuhan, China; PP-83 microelectrode puller was purchased from Narrishage, Japan. METHODS： The parietal somatosensory cortical neurons were acutely dissociated, and the modulation of acetylcholine （0.1, 1, 10, 100 μmol/L） on transient outward potassium channel was recorded using the whole-cell patch-clamp technique. MAIN OUTCOME MEASURES： Influence of acetylcholine on transient outward potassium current, potassium channel activation, and inactivation. RESULTS： The inhibitory effect of acetylcholine on transient outward potassium current was dose- and voltage-dependent （P 〈 0.01）. Acetylcholine was found to significantly affect the activation process of transient outward potassium current, i.e., the activation curve of transient outward potassium current was left-shifted, while the inactivation curve was shifted to hyperpolarization. Acetylcholine significantly prolonged the time constant of recovery from inactivation of transient outward potassium current （P 〈 0.01）. CONCLUSION： These results suggest that acetylcholine inhibits transient outward potassium current by regulating activation and inactivation processes of the transient outward potassium channel.

关键词： transient outward potassium channel whole-cell recording acetylcholine cortex

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Spontaneous firing properties of rat medial vestibular nucleus neurons in brain slices by infrared visual patch clamp technique

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Frontiers of Medicine 2008年第3期2卷 264-268页

作者： Jiao XIA Weijia KONG Yun ZHU Yan ZHOU Yu ZHANG Changkai GUO Department of Otorhinolaryngology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

Domestic application of infrared patch clamp techniques on brain slices is limited.The key of the tech-nique is to prepare high-quality brain slices.The present paper describes the preparation procedure of brainstem slices and the spontaneous firing properties of rat medial vestibular nucleus(MVN)neurons.By infrared differ-ential interference contrast technique,neurons of rat MVN were visualized directly at the depth of 50–100 mm underneath the surface of slices.Firing activities of MVN neurons were recorded by the whole-cell patch clamp technique in artificial cerebrospinal fluid(ACSF)and low Ca^(2+)-high Mg^(2+) fluid.The firing mode was more irregular and depressive in low Ca^(2+)-high Mg2+fluid than in ACSF.According to the averaged waveform of action potentials,cells were classified as the neurons with mono-phasic after-hyperpolarization potential(AHP),and the neurons with biphasic AHP.The resting membrane potential(RMP),input resistance(Rin)and membrane capacitance(Cm)of neurons were recorded and com-pared between groups.With infrared videomicroscopy,patch clamp recordings could be made under direct obser-vation in freshly prepared brainstem slices.The discharge activities of MVN neurons were spontaneous and the fir-ing mode was modulated by extracellular calcium concen-tration.The basic membrane properties of two types of neurons were not significantly different,while the differ-ences in waveform might play a role in the segregation between tonic and kinetic cells.

关键词： vestibular nucleus,medial brainstem slices infrared videomicroscopy whole-cell recording patch-clamp techniques spontaneous firing

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Glial Control of Depressive-like Behaviors

Glial Control of Depressive-like Behaviors

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中国神经科学学会第十二届全国学术会议

作者： Tian-Ming Gao Institutes of Mental Health Southern Medical University

Objective Major Depression is a common psychiatric disorder that affects over 120 million people worldwide, with a lifetime prevalence of 10% to 15%. Our previous study indicates that deficiency of astrocyte-derived ATP in the medial prefrontal cortex（m PFC） modulates depressive-like behaviors. However, the molecular mechanisms are not fully understood. Astrocytes release gliotransmitters to modulate synaptic activity, and alterations in excitation/inhibition balance may give rise to psychiatric disorders. Thus, the present study is to explore whether astrocytes modulate depressive-like behaviors through regulating synaptic activity. Methods Chronic social defeat stress was used to establish mouse model of depression. whole-cell recordings of inhibitory and excitatory synaptic currents were performed in pyramidal neurons in the m PFC. Pharmacological and genetic methods were used to verify the involvement of astrocytederived ATP and receptor subtypes. Retrograde labeling was applied to identify potential neural circuits involved. Results（1） Inhibitory, but not excitatory synaptic activities were impaired in depressed mice due to a presynaptic mechanism.（2） Perfusion of ATP could reverse the alterations of inhibitory synaptic activities in susceptible mice, without affecting excitatory synaptic activities.（3） IP3 R2 knockout mice, in which ATP release in astrocytes was specifically reduced, displayed similar impairments in synaptic transmission as in depressed mice and the reduced GABAergic synaptic activities were also rescued by application of ATP.（4） P2 X2 receptors mediated ATP modulation of GABAergic synaptic transmission.（5） Inhibitory synaptic activities on LHb-projecting pyramidal neurons in the m PFC were significantly decreased. Conclusion Astrocytederived ATP may modulate depressive-like behaviors through GABAergic inhibition in the mPFC.

关键词： Astrocytes medial prefrontal cortex major depression whole-cell recording synaptic transmission

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Astrocyte-mediated enhancement of neuronal excitability during spreading depolarization

Astrocyte-mediated enhancement of neuronal excitability duri...

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中国神经科学学会第十二届全国学术会议

作者： Dong Chuan WU Rita Yu-Tzu CHEN Ting-Chun CHENG Hong-Sha XUE Yu-Ying MEI Yao-Chang CHIANG Ning ZHOU Graduate Institute of Biomedical Sciences China Medical University Translational Medicine Research Center China Medical University Hospital Center for Drug Abuse and Addiction China Medical University Hospital China Medical University

Objective: Spreading depolarization(SD) is a pathophysiological phenomenon characterized by propagating waves of profound neuronal and glial depolarization in central nervous system grey matter. Although SD is primarily mediated by neurons with a subsequent astrocytic response, it remains unclear how astrocytic activity is modulated after SD. The present study aims to investigate whether altered astrocyte signalling contributes to neuronal excitability during the recovery of SD. Methods: Astrocyte somatic Ca changes were monitored with Rhod-2 by two-photon laser scanning microscopy. Slow inward currents(SICs) were recorded by whole-cell patch clamp recordings. Results: We found that after the concurrent Ca wave, SD enhanced astrocytic activity by promoting a secondary period of Ca oscillations. Different from the Ca wave that occurred concurrently with SD, the Ca oscillations are mediated by activation of GABA receptors and 1,4,5-trisphosphate(IP) receptors. Furthermore, SD increased the number of NMDA receptor-mediated SICs in CA1 pyramidal neurons. The astrocytic Ca oscillations and neuronal SICs had a similar time course. The frequency of SD-induced SICs was reduced by blockade of GABAB receptors or by limiting Ca efflux from the ER. Selective inhibition of astrocytic Ca signals by dialysis of BAPTA into astrocytes or by knocking out the astrocytic type of IP3 receptors suppressed SICs after SD. Injection of the SICs in pyramidal neurons evoked bursting of action potentials resembling to the SD-induced epileptiform activity. Therefore, we conclude that SD enhances the astrocyte Ca signals and further promotes gliotransmission and neuronal excitability.

关键词： Spreading depolarization astrocytes Ca2+ oscillations slow inward currents whole-cell recording

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Requirement for activation of glycine site of NMDA receptors in the rostral anterior cingulate cortex in cat pain-related negative affect

Requirement for activation of glycine site of NMDA receptors...

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中国神经科学学会第六届学术会议暨学会成立十周年庆祝大会

作者： Ren Wen-hua, Guo Ji-dong, Cao Hong, Ru-Rong ji, Zhao Zhi-qi, Zhang Yu-qiu Institute of Neurobiology, Fudan University, Shanghai 200433, China Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China: Pain Research Center, Dept. of Anesthesiology, Brigham and Women''s Hospital, Harvard Medical School, Boston, MA 02115, USA

＜正＞Emotional distress is the most disruptive and undesirable feature of painful experiences. More and more clinical observations indicate the patients with chronic pain suffer from as much emotional disturbance as pain sensation per se. Physiological arousal and hypersensitivity to pain cause varying severities and qualities of negative affect, such as fear, anxiety, anger, aversion, and depression, even tendency of suicide, and these negative affective states in turn enhance pain perception. Previous study suggests that the activation of NMDA receptors in the rostrol cingulated cortex （rACC） may involve in pain related aversive emotion acquisition. Combination of a rat formalin-induced conditioned place

关键词： pain-related negative emotion conditioned place avoidance （CPA） glycine site of NMDA receptors whole-cell recording rostral anterior cingulate cortex （ACC） rat

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Synaptic mechanisms of direction selectivity in primary auditory cortex

Synaptic mechanisms of direction selectivity in primary audi...

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中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Mu-Ming POO Institute of Neuroscience and State Key Laboratory of Neuroscience Shanghai Institutes for Biological SciencesChinese Academy of Sciences Division of Neurobiology Department of Molecular and Cell Biology ＆ Helen Wills Neuroscience InstituteUniversity of California

＜正＞Objective Frequency modulation（FM） is a prominent feature in animal vocalization and human speech. Although many neurons in the auditory cortex are known to be selective for FM direction,the synaptic mechanisms underlying this selectivity are not well understood.Previous studies of both visual and auditory neurons have suggested two general mechanisms for direction selectivity:（1） differential delays of excitatory inputs across the spatial/spectral receptive field and（2） spatial/spectral offset between excitatory and inhibitory inputs.In this study,we have examined the contributions of both mechanisms to FM direction selectivity in rat primary auditory cortex（A1）.Methods The excitatory and inhibitory synaptic inputs to each cortical neuron were measured by in vivo whole-cell recording.The spectrotemporal receptive field（STRF） of each type of inputs was mapped with random tone pips and compared with direction selectivity of the neuron measured with FM stimuli.Results We found that（1） Direction selectivity measured from the spiking output is significantly stronger than that based on the subthreshold membrane potential changes.（2） Both the differential delays of excitatory inputs and the spectral offset between excitation and inhibition are positively correlated with direction selectivity of the neuron.Conclusion Our results suggest that both synaptic mechanisms are likely to contribute to FM direction selectivity at different extents in Al,and the selectivity is further sharpened by the nonlinear transformation mechanism from subthreshold membrane potentials to the spike.

关键词： direction selectivity synaptic mechanism spectrotemporal receptive field primary auditory cortex whole-cell recording rat

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Synthesis and characterization of huwentoxin-Ⅳ,a neurotoxin inhibiting central neuronal sodium channels

Synthesis and characterization of huwentoxin-Ⅳ,a neurotoxin...

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第八届中国生物毒素学术研讨会

作者： Xuan Luo# Yucheng Xiao# Fang Kuang Meichun Deng Meichi Wang Songping Liang~* Life Sciences College,Hunan Normal University,Changsha,Hunan 410081,P.R.China

Our previous work demonstrated that huwentoxin-Ⅳ was an inhibitor cystine knot peptide from Chinese tarantula Ornithoctonus huwena venom that blocked tetrodotoxin-sensitive voltage-gated sodium channels from mammalian sensory neurons.However,the actions of the neurotoxin on central neuronal sodium channels remain unknown.In this study,we chemically synthesized native huwentoxin-Ⅳ and found that sodium channel isoforms from rat hippocampus neurons were also sensitive to native and synthetic toxins,but the toxin binding affinity (IC50 .4 μM) was 12-fold lower than to peripheral isoforms.The blockade by huwentoxin-Ⅳ could be reversed by strong depolarization due to the dissociation of toxin-channel complex as observed for receptor site 3 toxins.Moreover,small unilamellar vesicles binding assays showed that in contrast to ProTx-Ⅱ from the tarantula Thrixopelma pruriens,huwentoxin-Ⅳ lacked the ability to partition into the negatively charged and neutral phospholipid bilayer of artificial membranes.These findings indicated that huwentoxin-Ⅳ was a sodium channel antagonist preferentially targeting peripheral isoforms via a mechanism quite different from ProTx-Ⅱ.

关键词： spider toxin sodium channel whole-cell recording hippocampus

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