17 results on '"Shujiang Shang"'
Search Results
2. Intracellular TRPA1 mediates Ca2+ release from lysosomes in dorsal root ganglion neurons
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Li Wang, Lei Sun, Zhuan Zhou, Bing Liu, Qihui Wu, Xiaoyu Zhang, Yuan Wang, Bin Liu, Shujiang Shang, Chen Zhang, Xi Wu, Michael X. Zhu, Xinghua Feng, Changhe Wang, Desheng Zhu, Sasa Teng, Rong Huang, Meiqin Hu, Liu Tao, Fukang Zhang, Lianghong Zheng, Zuying Chai, Huadong Xu, Yeshi Wang, and Feipeng Zhu
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0301 basic medicine ,Calcitonin gene-related peptide ,Biology ,Article ,Exocytosis ,03 medical and health sciences ,Transient receptor potential channel ,0302 clinical medicine ,Dorsal root ganglion ,medicine ,Extracellular ,Research Articles ,Calcium signaling ,Vesicle ,food and beverages ,Cell Biology ,3. Good health ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Biochemistry ,psychological phenomena and processes ,030217 neurology & neurosurgery ,Intracellular - Abstract
The temperature-sensitive TRP channel, TRPA1, is known to mediate Na+ and Ca2+ influx at the plasma membrane of sensory neurons. In this study, the authors show that TRPA1 is also present on the lysosomal membrane and mediates lysosome Ca2+ release in dorsal root ganglion neurons., Transient receptor potential A1 (TRPA1) is a nonselective cation channel implicated in thermosensation and inflammatory pain. In this study, we show that TRPA1 (activated by allyl isothiocyanate, acrolein, and 4-hydroxynonenal) elevates the intracellular Ca2+ concentration ([Ca2+]i) in dorsal root ganglion (DRG) neurons in the presence and absence of extracellular Ca2+. Pharmacological and immunocytochemical analyses revealed the presence of TRPA1 channels both on the plasma membrane and in endolysosomes. Confocal line-scan imaging demonstrated Ca2+ signals elicited from individual endolysosomes (“lysosome Ca2+ sparks”) by TRPA1 activation. In physiological solutions, the TRPA1-mediated endolysosomal Ca2+ release contributed to ∼40% of the overall [Ca2+]i rise and directly triggered vesicle exocytosis and calcitonin gene-related peptide release, which greatly enhanced the excitability of DRG neurons. Thus, in addition to working via Ca2+ influx, TRPA1 channels trigger vesicle release in sensory neurons by releasing Ca2+ from lysosome-like organelles.
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- 2016
3. An Excitatory Neural Assembly Encodes Working Memory in the Prefrontal Cortex
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Yong-Jie Wang, Yaxuan Cui, Liecheng Wang, Zilong Qiu, Yangzhen Wang, Desheng Zhu, Yunbo Liu, Min-Hua Luo, Feng Su, Xunli Wang, Peng Cao, Shiming Tang, Chen Zhang, Xiang-Yao Li, Chaojuan Yang, Yonglu Tian, Shujiang Shang, and Peijiang Yuan
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Elementary cognitive task ,medicine.anatomical_structure ,Schizophrenia ,Working memory ,Mechanism (biology) ,medicine ,Excitatory postsynaptic potential ,Sensory system ,Sensory cortex ,Biology ,Prefrontal cortex ,medicine.disease ,Neuroscience - Abstract
Working memory (WM) is crucial for animals to execute cognitive tasks that require short-term storage and manipulation of sensory information in the order of seconds to minutes. WM is strongly correlated with human intelligence and deficits in this process lead to many psychiatric disorders, including Alzheimer's disease (AD), schizophrenia and autism. The proper functioning of WM involves multiple brain areas, including the sensory cortex and prefrontal cortex. Persistent or recurrent neural activity within these brain areas, together with neural oscillations among these brain areas, is known to encode the WM. However, the coding mechanisms of WM at the microcircuitry level still remain elusive. In this study, we performed two-photon imaging on behaving mice to monitor the activity of neuronal microcircuitry. We also revealed a neuronal subpopulation in the medial prefrontal cortex (mPFC) that exhibited emergent properties in a context-dependent manner underlying a WM-like behavior paradigm. These neuronal subpopulations are exclusively composed of excitatory neurons and mainly represent a group of neurons with more functional connections within the mPFC microcircuitry. This type of microcircuitry plasticity was maintained in minutes and was absent in an animal model of AD. Thus, these results identify a novel functional coding mechanism that relies on the emergent behavior of a functional-defined neuronal assembly to encode WM.
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- 2018
4. Loss of FMRP Impaired Hippocampal Long-Term Plasticity and Spatial Learning in Rats
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Yonglu Tian, Chaojuan Yang, Shujiang Shang, Yijun Cai, Xiaofei Deng, Jian Zhang, Feng Shao, Desheng Zhu, Yunbo Liu, Guiquan Chen, Jing Liang, Qiang Sun, Zilong Qiu, and Chen Zhang
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0301 basic medicine ,congenital, hereditary, and neonatal diseases and abnormalities ,hippocampus ,Morris water navigation task ,AMPA receptor ,lcsh:RC321-571 ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,medicine ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,Molecular Biology ,Original Research ,Macroorchidism ,spatial learning ,Long-term potentiation ,medicine.disease ,FMR1 ,long-term plasticity ,Fragile X syndrome ,030104 developmental biology ,medicine.anatomical_structure ,intellectual disability ,Schaffer collateral ,Synaptic plasticity ,FXS ,Psychology ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Fragile X syndrome (FXS) is a neurodevelopmental disorder caused by mutations in the FMR1 gene that inactivate expression of the gene product, the fragile X mental retardation 1 protein (FMRP). In this study, we used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology to generate Fmr1 knockout (KO) rats by disruption of the fourth exon of the Fmr1 gene. Western blotting analysis confirmed that the FMRP was absent from the brains of the Fmr1 KO rats (Fmr1exon4-KO). Electrophysiological analysis revealed that the theta-burst stimulation (TBS)–induced long-term potentiation (LTP) and the low-frequency stimulus (LFS)–induced long-term depression (LTD) were decreased in the hippocampal Schaffer collateral pathway of the Fmr1exon4-KO rats. Short-term plasticity, measured as the paired-pulse ratio, remained normal in the KO rats. The synaptic strength mediated by the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) was also impaired. Consistent with previous reports, the Fmr1exon4-KO rats demonstrated an enhanced 3,5-dihydroxyphenylglycine (DHPG)–induced LTD in the present study, and this enhancement is insensitive to protein translation. In addition, the Fmr1exon4-KO rats showed deficits in the probe trial in the Morris water maze test. These results demonstrate that deletion of the Fmr1 gene in rats specifically impairs long-term synaptic plasticity and hippocampus-dependent learning in a manner resembling the key symptoms of FXS. Furthermore, the Fmr1exon4-KO rats displayed impaired social interaction and macroorchidism, the results consistent with those observed in patients with FXS. Thus, Fmr1exon4-KO rats constitute a novel rat model of FXS that complements existing mouse models.
- Published
- 2017
5. An Excitatory Neural Assembly Encodes Short-Term Memory in the Prefrontal Cortex
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Chen Zhang, Yonglu Tian, Shiming Tang, Fei Zhao, Yong-Jie Wang, Liecheng Wang, Desheng Zhu, Wen-Bo Zeng, Xiang-Yao Li, Zilong Qiu, Hao Li, Yunbo Liu, Peng Cao, Feng Su, Yaxuan Cui, Hai-Fei Jiang, Xunli Wang, Yangzhen Wang, Wei Xiong, Shujiang Shang, Jizong Zhao, Chaojuan Yang, Min-Hua Luo, and Peijiang Yuan
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0301 basic medicine ,Male ,short-term memory ,Short-term memory ,Pain ,Prefrontal Cortex ,Mice, Transgenic ,Biology ,Somatosensory system ,General Biochemistry, Genetics and Molecular Biology ,Extinction, Psychological ,03 medical and health sciences ,Neural activity ,0302 clinical medicine ,Animals ,Prefrontal cortex ,lcsh:QH301-705.5 ,Neurons ,Neuronal Plasticity ,Behavior, Animal ,Working memory ,Long-term potentiation ,Entorhinal cortex ,Mice, Inbred C57BL ,030104 developmental biology ,Memory, Short-Term ,Sound ,lcsh:Biology (General) ,nervous system ,Organ Specificity ,Excitatory postsynaptic potential ,Nerve Net ,Alzheimer’s disease ,Neuroscience ,microcircuitry ,030217 neurology & neurosurgery - Abstract
Summary: Short-term memory (STM) is crucial for animals to hold information for a small period of time. Persistent or recurrent neural activity, together with neural oscillations, is known to encode the STM at the cellular level. However, the coding mechanisms at the microcircuitry level remain a mystery. Here, we performed two-photon imaging on behaving mice to monitor the activity of neuronal microcircuitry. We discovered a neuronal subpopulation in the medial prefrontal cortex (mPFC) that exhibited emergent properties in a context-dependent manner underlying a STM-like behavior paradigm. These neuronal subpopulations exclusively comprise excitatory neurons and mainly represent a group of neurons with stronger functional connections. Microcircuitry plasticity was maintained for minutes and was absent in an animal model of Alzheimer’s disease (AD). Thus, these results point to a functional coding mechanism that relies on the emergent behavior of a functionally defined neuronal assembly to encode STM.
- Published
- 2017
6. Extracellular Ca2+ per se inhibits quantal size of catecholamine release in adrenal slice chromaffin cells
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Changhe Wang, Shirong Wang, Liu Tao, Xinjiang Kang, Hui Zheng, Shujiang Shang, Qihui Wu, Wei Liu, Wei Xiong, Yeshi Wang, Xiaoyu Zhang, Zhuan Zhou, Quanfeng Zhang, Huadong Xu, Bin Liu, and Lianghong Zheng
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Physiology ,Vesicle ,Kinetics ,chemistry.chemical_element ,Cell Biology ,Calcium ,Biology ,Amperometry ,Cell biology ,medicine.anatomical_structure ,chemistry ,Chromaffin cell ,Catecholamine ,medicine ,Extracellular ,Molecular Biology ,Intracellular ,medicine.drug - Abstract
Classic calcium hypothesis states that depolarization-induced increase in intracellular Ca(2+) concentration ([Ca(2+)]i) triggers vesicle exocytosis by increasing vesicle release probability in neurons and neuroendocrine cells. The extracellular Ca(2+), in this calcium hypothesis, serves as a reservoir of Ca(2+) source. Recently we find that extracellular Ca(2+)per se inhibits the [Ca(2+)]i dependent vesicle exocytosis, but it remains unclear whether quantal size is regulated by extracellular, or intracellular Ca(2+) or both. In this work we showed that, in physiological condition, extracellular Ca(2+) per se specifically inhibited the quantal size of single vesicle release in rat adrenal slice chromaffin cells. The extracellular Ca(2+) in physiological concentration (2.5 mM) directly regulated fusion pore kinetics of spontaneous quantal release of catecholamine. In addition, removal of extracellular Ca(2+) directly triggered vesicle exocytosis without eliciting intracellular Ca(2+). We propose that intracellular Ca(2+) and extracellular Ca(2+)per se cooperately regulate single vesicle exocytosis. The vesicle release probability was jointly modulated by both intracellular and extracellular Ca(2+), while the vesicle quantal size was mainly determined by extracellular Ca(2+) in chromaffin cells physiologically.
- Published
- 2014
7. Effect of Cocaine on Ion Channels and Glutamatergic EPSCs in Noradrenergic Locus Coeruleus Neurons
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H. Yang, Changhe Wang, Bing Liu, Shirong Wang, Xiaoyu Zhang, S. T. Kuo, Li Mingli, Huadong Xu, L. N. Liu, M. Y. Song, Feipeng Zhu, Shujiang Shang, Li Zhou, Ruiying Jiao, Bo Zhang, Sasa Teng, Zhuan Zhou, Wei Liu, Xinjiang Kang, and Zhi-Qing David Xu
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medicine.medical_specialty ,Epinephrine ,Glutamic Acid ,Galanin ,Neurotransmission ,Ion Channels ,Rats, Sprague-Dawley ,Cellular and Molecular Neuroscience ,Glutamatergic ,Cocaine ,Neuromodulation ,Internal medicine ,medicine ,Animals ,Neurons ,Voltage-gated ion channel ,Chemistry ,Sodium channel ,Sodium ,Excitatory Postsynaptic Potentials ,General Medicine ,Rats ,Endocrinology ,medicine.anatomical_structure ,nervous system ,Potassium ,Excitatory postsynaptic potential ,Locus coeruleus ,Calcium ,Locus Coeruleus ,Neuroscience - Abstract
The locus coeruleus (LC) is an important brainstem area involved in cocaine addiction. However, evidence to elucidate how cocaine modulates the activity of LC neurons remains incomplete. Here, we performed whole recordings in brain slices to evaluate the effects of cocaine on the sodium (Na(+)), potassium (K(+)), calcium (Ca(2+)) channels, and glutamatergic synaptic transmission in the locus coeruleus neurons. Local application of cocaine significantly and reversibly reduced the spontaneous firing rate but did not affect action potential amplitude, rising time, decay time, or half width of noradrenergic locus coeruleus neurons. Moreover, cocaine attenuated the sodium current but did not affect potassium and calcium currents. The N-methyl-D-aspartate receptor mediated excitatory postsynaptic currents were reduced by neuropeptide galanin but not cocaine. All those data demonstrate that cocaine has inhibitory effect on the spontaneous activities and sodium current in locus coeruleus neurons. Therefore, neuromodulation of sodium channel in locus coeruleus neurons may play an important role in drug addiction.
- Published
- 2013
8. Calcium Triggers Exocytosis from Two Types of Organelles in a Single Astrocyte
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Weiping Han, Sasa Teng, Li Wang, Claire Xi Zhang, Changhe Wang, Lianghong Zheng, Shujiang Shang, Ning Guo, Yeshi Wang, Liu Tao, Lei Sun, Bin Liu, Yingfei Xiong, and Zhuan Zhou
- Subjects
Vesicle-Associated Membrane Protein 3 ,Vesicle-Associated Membrane Protein 2 ,Green Fluorescent Proteins ,Neurotoxins ,Glutamic Acid ,Neurotransmission ,Transfection ,Hippocampus ,Exocytosis ,Mice ,Tetanus Toxin ,Lysosomal-Associated Membrane Protein 1 ,Lysosome ,Glial Fibrillary Acidic Protein ,medicine ,Animals ,Humans ,Calcium Signaling ,Transport Vesicles ,Cells, Cultured ,Cellular compartment ,Chemistry ,General Neuroscience ,Vesicle ,Glutamate receptor ,Articles ,Cell biology ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Receptors, Glutamate ,Membrane protein ,Astrocytes ,Mutation ,Vesicular Glutamate Transport Protein 1 ,Calcium ,Lysosomes ,Astrocyte - Abstract
Astrocytes release a variety of signaling molecules including glutamate, D-serine, and ATP in a regulated manner. Although the functions of these molecules, from regulating synaptic transmission to controlling specific behavior, are well documented, the identity of their cellular compartment(s) is still unclear. Here we set out to study vesicular exocytosis and glutamate release in mouse hippocampal astrocytes. We found that small vesicles and lysosomes coexisted in the same freshly isolated or cultured astrocytes. Both small vesicles and lysosome fused with the plasma membrane in the same astrocytes in a Ca(2+)-regulated manner, although small vesicles were exocytosed more efficiently than lysosomes. Blockade of the vesicle glutamate transporter or cleavage of synaptobrevin 2 and cellubrevin (both are vesicle-associated membrane proteins) with a clostridial toxin greatly inhibited glutamate release from astrocytes, while lysosome exocytosis remained intact. Thus, both small vesicles and lysosomes contribute to Ca(2+)-dependent vesicular exocytosis, and small vesicles support glutamate release from astrocytes.
- Published
- 2011
9. Two distinct vesicle pools for depolarization-induced exocytosis in somata of dorsal root ganglion neurons
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Yeshi Wang, Liu Tao, Changhe Wang, Zhuan Zhou, Claire Xi Zhang, Wei Xiong, Shujiang Shang, Bin Liu, and Lianghong Zheng
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Physiology ,Chemistry ,Vesicle ,Stimulation ,Depolarization ,Hormone release ,Kiss-and-run fusion ,Exocytosis ,Cell biology ,chemistry.chemical_compound ,medicine.anatomical_structure ,nervous system ,Dorsal root ganglion ,medicine ,Neurotransmitter - Abstract
Non-technical summary Ca2+-regulated exocytosis is essential for neurotransmitter and hormone release. As well as this type of exocytosis, the somata of dorsal root ganglion (DRG) neurons also show Ca2+-independent but voltage-dependent exocytosis. It is unclear whether these two types of exocytosis use the same or different vesicle pools in DRG neurons. Here, we found that they were separable in response to the same stimulation in low external Ca2+ solution. Depletion of the Ca2+-dependent vesicle pool did not affect the Ca2+-independent but voltage-dependent exocytosis. These results show that DRG neurons exhibit two distinct types of exocytosis that use different vesicle pools.
- Published
- 2011
10. Rhesus Macaques Develop Metabolic Syndrome With Reversible Vascular Dysfunction Responsive to Pioglitazone
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Lifang Xue, Kaitao Li, Xiuqin Zhang, Chao Han, Zhentao Song, Rongli Zhang, Huiliang Zhang, Shujiang Shang, Hui Wang, Chuan-Yun Li, Jiaming Mao, Jue Wang, Susanne Raab, Tiangang Zhu, Wen Zheng, Na Liu, Heping Cheng, Yan Zhang, Paul L. Huang, Ning Hou, Yuli Liu, Yi Ding, and Elena Sebokova
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medicine.medical_specialty ,Fasting hyperinsulinemia ,Physiology ,Article ,Pathogenesis ,Insulin resistance ,Hyperinsulinism ,Physiology (medical) ,Internal medicine ,medicine ,Animals ,Hypoglycemic Agents ,Dyslipidemias ,Metabolic Syndrome ,Pioglitazone ,business.industry ,Metabolic disorder ,medicine.disease ,Macaca mulatta ,Obesity ,Disease Models, Animal ,Endocrinology ,Regional Blood Flow ,Obesity, Abdominal ,Hypertension ,Disease Progression ,Blood Vessels ,Thiazolidinediones ,Insulin Resistance ,Metabolic syndrome ,Cardiology and Cardiovascular Medicine ,business ,medicine.drug - Abstract
Background— The metabolic syndrome (MetS) is a constellation of clinical features that include central obesity, hypertension, atherogenic dyslipidemia, and insulin resistance. However, the concept remains controversial; it has been debated whether MetS represents nothing more than simultaneous co-occurrence of individual risk factors or whether there are common shared pathophysiological mechanisms that link the individual components. Methods and Results— To investigate the emergence of metabolic and cardiovascular components during the development of MetS, we identified MetS-predisposed animals (n=35) in a large population of rhesus macaques (Macaca mulatta, 12.7±2.9 years old, n=408), acclimated them to standardized conditions, and monitored the progression of individual component features over 18 months. In 18 MetS animals with recently developed fasting hyperinsulinemia, central obesity, hypertension, and atherogenic dyslipidemia, we found that individual metabolic and cardiovascular components track together during the transition from pre-MetS to onset of MetS; MetS was associated with a 60% impairment of flow-mediated dilation, establishing the mechanistic link with vascular dysfunction. Pioglitazone treatment (3 mg/kg body weight/d for 6 weeks), a peroxisome proliferator–activated receptor γ agonist, reversibly improved atherogenic dyslipidemia and insulin resistance and fully restored flow-mediated dilation with persistent benefits. Conclusions— Coemergence of metabolic and cardiovascular components during MetS progression and complete normalization of vascular dysfunction with peroxisome proliferator-activated receptor γ agonists suggest shared underlying mechanisms rather than separate processes, arguing for the benefit of early intervention of MetS components. Predictive nonhuman primate (NHP) models of MetS should be highly valuable in mechanistic and translational studies on the pathogenesis of MetS in relation to cardiovascular disease and diabetes mellitus.
- Published
- 2011
11. Imaging superoxide flash and metabolism-coupled mitochondrial permeability transition in living animals
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Xiuqin Zhang, Xing Zhang, Feng Gao, Shujiang Shang, Huaqiang Fang, Heping Cheng, Jianjie Ma, Shi-Qiang Wang, Jiejia Xu, Kunfu Ouyang, Yi Ding, Wanrui Zhang, Ju Chen, Ming Zheng, Xianhua Wang, Yao Xiao, Jing Chao Li, Xiao-Li Tian, Noah Weisleder, Wei Shang, Jingsong Zhou, Min Chen, Kaitao Li, and Wang Wang
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Mitochondrial ROS ,Mice, Transgenic ,Mitochondrion ,Mitochondrial Membrane Transport Proteins ,Mice ,chemistry.chemical_compound ,Mitochondrial membrane transport protein ,Bacterial Proteins ,Superoxides ,medicine ,Animals ,Insulin ,Muscle, Skeletal ,Molecular Biology ,chemistry.chemical_classification ,Reactive oxygen species ,Microscopy, Confocal ,biology ,Mitochondrial Permeability Transition Pore ,Superoxide ,Skeletal muscle ,Cell Biology ,Mitochondria ,Cell biology ,Luminescent Proteins ,Glucose ,medicine.anatomical_structure ,chemistry ,Mitochondrial permeability transition pore ,biology.protein ,Original Article ,Signal transduction ,Energy Metabolism ,Reactive Oxygen Species ,Signal Transduction - Abstract
The mitochondrion is essential for energy metabolism and production of reactive oxygen species (ROS). In intact cells, respiratory mitochondria exhibit spontaneous "superoxide flashes", the quantal ROS-producing events consequential to transient mitochondrial permeability transition (tMPT). Here we perform the first in vivo imaging of mitochondrial superoxide flashes and tMPT activity in living mice expressing the superoxide biosensor mt-cpYFP, and demonstrate their coupling to whole-body glucose metabolism. Robust tMPT/superoxide flash activity occurred in skeletal muscle and sciatic nerve of anesthetized transgenic mice. In skeletal muscle, imaging tMPT/superoxide flashes revealed labyrinthine three-dimensional networks of mitochondria that operate synchronously. The tMPT/superoxide flash activity surged in response to systemic glucose challenge or insulin stimulation, in an apparently frequency-modulated manner and involving also a shift in the gating mode of tMPT. Thus, in vivo imaging of tMPT-dependent mitochondrial ROS signals and the discovery of the metabolism-tMPT-superoxide flash coupling mark important technological and conceptual advances for the study of mitochondrial function and ROS signaling in health and disease.
- Published
- 2011
12. Calcium influx activates adenylyl cyclase 8 for sustained insulin secretion in rat pancreatic beta cells
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Xi Wu, Li Wang, Sasa Teng, Yeshi Wang, Liangyi Chen, Bin Liu, Lijun Yao, Zhuan Zhou, Xiao Yu, Meiqin Hu, Jin Zhang, Qing Li, Shujiang Shang, Jochen Lang, Huadong Xu, Xiaoyu Zhang, Changhe Wang, Xinjiang Kang, Qihui Wu, Quanfeng Zhang, Lianghong Zheng, Jing Su, Haiqiang Dou, Matthieu Raoux, and Yi Wu
- Subjects
medicine.medical_specialty ,Vesicle fusion ,Patch-Clamp Techniques ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Biology ,Exocytosis ,ADCY10 ,Adenylyl cyclase ,chemistry.chemical_compound ,Internal medicine ,Insulin-Secreting Cells ,Internal Medicine ,medicine ,Cyclic AMP ,Animals ,Patch clamp ,Rats, Wistar ,Protein kinase A ,Cells, Cultured ,Insulin ,Cyclic AMP-Dependent Protein Kinases ,Cell biology ,Rats ,Disease Models, Animal ,Endocrinology ,chemistry ,Calcium ,Signal transduction ,Adenylyl Cyclases ,Signal Transduction - Abstract
Insulin is a key metabolic regulator in health and diabetes. In pancreatic beta cells, insulin release is regulated by the major second messengers Ca(2+) and cAMP: exocytosis is triggered by Ca(2+) and mediated by the cAMP/protein kinase A (PKA) signalling pathway. However, the causal link between these two processes in primary beta cells remains undefined.Time-resolved confocal imaging of fluorescence resonance energy transfer signals was performed to visualise PKA activity, and combined membrane capacitance recordings were used to monitor insulin secretion from patch-clamped rat beta cells.Membrane depolarisation-induced Ca(2+) influx caused an increase in cytosolic PKA activity via activating a Ca(2+)-sensitive adenylyl cyclase 8 (ADCY8) subpool. Glucose stimulation triggered coupled Ca(2+) oscillations and PKA activation. ADCY8 knockdown significantly reduced the level of depolarisation-evoked PKA activation and impaired replenishment of the readily releasable vesicle pool. Pharmacological inhibition of PKA by two inhibitors reduced depolarisation-induced PKA activation to a similar extent and reduced the capacity for sustained vesicle exocytosis and insulin release.Our findings suggest that depolarisation-induced Ca(2+) influx plays dual roles in regulating exocytosis in rat pancreatic beta cells by triggering vesicle fusion and replenishing the vesicle pool to support sustained insulin release. Therefore, Ca(2+) influx may be important for glucose-stimulated insulin secretion.
- Published
- 2014
13. Modulation of dopamine release in the striatum by physiologically relevant levels of nicotine
- Author
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Shirong Wang, Bin Liu, Li Zhou, Sasa Teng, Ruiying Jiao, Feipeng Zhu, Shujiang Shang, Haiqiang Dou, Wei Liu, Zuo Panli, Qihui Wu, Li Wang, Li Mingli, Zhuan Zhou, Xiaoyu Zhang, Lianghong Zheng, Changhe Wang, Xinjiang Kang, and Huadong Xu
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Nicotine ,Dopamine ,General Physics and Astronomy ,Mice, Transgenic ,Stimulation ,Striatum ,In Vitro Techniques ,Pharmacology ,Optogenetics ,General Biochemistry, Genetics and Molecular Biology ,Mice ,medicine ,Animals ,Axon ,Multidisciplinary ,Chemistry ,Dopaminergic ,General Chemistry ,Corpus Striatum ,Electric Stimulation ,medicine.anatomical_structure ,nervous system ,Cholinergic ,medicine.drug - Abstract
Striatal dopamine (DA) release can be independently triggered not only by action potentials (APs) in dopaminergic axons but also APs in cholinergic interneurons (ChIs). Nicotine causes addiction by modulating DA release, but with paradoxical findings. Here, we investigate how physiologically relevant levels of nicotine modulate striatal DA release. The optogenetic stimulation of ChIs elicits DA release, which is potently inhibited by nicotine with an IC50 of 28 nM in the dorsal striatum slice. This ChI-driven DA release is predominantly mediated by α6β2* nAChRs. Local electrical stimulus (Estim) activates both dopaminergic axons and ChIs. Nicotine does not affect the AP(DA)-dependent DA release (AP(DA), AP of dopaminergic axon). During burst Estim, nicotine permits the facilitation of DA release by prevention of DA depletion. Our work indicates that cholinergic stimulation-induced DA release is profoundly modulated by physiologically relevant levels of nicotine and resolves the paradoxical observation of nicotine's effects on striatal DA release.
- Published
- 2014
14. Extracellular Ca²⁺ per se inhibits quantal size of catecholamine release in adrenal slice chromaffin cells
- Author
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Shujiang, Shang, Changhe, Wang, Bin, Liu, Qihui, Wu, Quanfeng, Zhang, Wei, Liu, Lianghong, Zheng, Huadong, Xu, Xinjiang, Kang, Xiaoyu, Zhang, Yeshi, Wang, Hui, Zheng, Shirong, Wang, Wei, Xiong, Tao, Liu, and Zhuan, Zhou
- Subjects
Catecholamines ,Chromaffin Cells ,Secretory Vesicles ,Adrenal Glands ,Animals ,Calcium ,Rats, Wistar ,Cells, Cultured ,Exocytosis ,Rats - Abstract
Classic calcium hypothesis states that depolarization-induced increase in intracellular Ca(2+) concentration ([Ca(2+)]i) triggers vesicle exocytosis by increasing vesicle release probability in neurons and neuroendocrine cells. The extracellular Ca(2+), in this calcium hypothesis, serves as a reservoir of Ca(2+) source. Recently we find that extracellular Ca(2+)per se inhibits the [Ca(2+)]i dependent vesicle exocytosis, but it remains unclear whether quantal size is regulated by extracellular, or intracellular Ca(2+) or both. In this work we showed that, in physiological condition, extracellular Ca(2+) per se specifically inhibited the quantal size of single vesicle release in rat adrenal slice chromaffin cells. The extracellular Ca(2+) in physiological concentration (2.5 mM) directly regulated fusion pore kinetics of spontaneous quantal release of catecholamine. In addition, removal of extracellular Ca(2+) directly triggered vesicle exocytosis without eliciting intracellular Ca(2+). We propose that intracellular Ca(2+) and extracellular Ca(2+)per se cooperately regulate single vesicle exocytosis. The vesicle release probability was jointly modulated by both intracellular and extracellular Ca(2+), while the vesicle quantal size was mainly determined by extracellular Ca(2+) in chromaffin cells physiologically.
- Published
- 2014
15. Importing, caring, breeding, genotyping, and phenotyping a genetic mouse in a Chinese university
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Xiaoyu Zhang, D. S. Zhu, Wei Liu, Shirong Wang, Xi Wu, Yeshi Wang, Meiqin Hu, Li Zhou, Q. H. Wu, Feipeng Zhu, Sasa Teng, Zhuan Zhou, Li Wang, S. T. Kuo, Quanfeng Zhang, Huadong Xu, Haiqiang Dou, Lianghong Zheng, Zuying Chai, Shujiang Shang, Zuo Panli, Xinjiang Kang, N. Guo, Changhe Wang, Z. L. Xie, Chen Zhang, Bing Liu, and L. N. Liu
- Subjects
Genetics ,Genetically modified mouse ,Mice, Knockout ,China ,Animal Welfare (journal) ,Animal Care Committees ,Genotyping Techniques ,Universities ,Laboratory mouse ,General Medicine ,Biology ,Breeding ,Cellular and Molecular Neuroscience ,Mice ,Laboratory Animal Science ,Knockout mouse ,Animals ,BioHazard ,Genotyping ,Dynamin I - Abstract
The genetic manipulation of the laboratory mouse has been well developed and generated more and more mouse lines for biomedical research. To advance our science exploration, it is necessary to share genetically modified mouse lines with collaborators between institutions, even in different countries. The transfer process is complicated. Significant paperwork and coordination are required, concerning animal welfare, intellectual property rights, colony health status, and biohazard. Here, we provide a practical example of importing a transgenic mice line, Dynamin 1 knockout mice, from Yale University in the USA to Perking University in China for studying cell secretion. This example including the length of time that required for paper work, mice quarantine at the receiving institution, and expansion of the mouse line for experiments. The procedure described in this paper for delivery live transgenic mice from USA to China may serve a simple reference for transferring mouse lines between other countries too.
- Published
- 2013
16. Thiopental-induced insulin secretion via activation of IP3-sensitive calcium stores in rat pancreatic β-cells
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Yunfei Xu, Hai-Qiang Dou, Xiao Yu, Chao-chao Chen, Shujiang Shang, Jin-Peng Sun, Lianghong Zheng, Shu Guo, Iain C. Bruce, and Zhuan Zhou
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Boron Compounds ,medicine.medical_specialty ,Patch-Clamp Techniques ,Potassium Channels ,Physiology ,chemistry.chemical_element ,Inositol 1,4,5-Trisphosphate ,Calcium ,Carbohydrate metabolism ,Biology ,Membrane Potentials ,Cell membrane ,Internal medicine ,Insulin-Secreting Cells ,Insulin Secretion ,medicine ,Animals ,Inositol 1,4,5-Trisphosphate Receptors ,Insulin ,Patch clamp ,Estrenes ,Rats, Wistar ,Thiopental ,Insulin secretion ,Calcium metabolism ,Heparin ,Ryanodine ,Cell Biology ,Pyrrolidinones ,Rats ,medicine.anatomical_structure ,Endocrinology ,Glucose ,chemistry ,Thapsigargin ,Anesthetics, Intravenous ,Insulin metabolism - Abstract
While glucose-stimulated insulin secretion depends on Ca2+ influx through voltage-gated Ca2+ channels in the cell membrane of the pancreatic β-cell, there is also ample evidence for an important role of intracellular Ca2+ stores in insulin secretion, particularly in relation to drug stimuli. We report here that thiopental, a common anesthetic agent, triggers insulin secretion from the intact pancreas and primary cultured rat pancreatic β-cells. We investigated the underlying mechanisms by measurements of whole cell K+ and Ca2+ currents, membrane potential, cytoplasmic Ca2+ concentration ([Ca2+]i), and membrane capacitance. Thiopental-induced insulin secretion was first detected by enzyme-linked immunoassay, then further assessed by membrane capacitance measurement, which revealed kinetics distinct from glucose-induced insulin secretion. The thiopental-induced secretion was independent of cell membrane depolarization and closure of ATP-sensitive potassium (KATP) channels. However, accompanied by the insulin secretion stimulated by thiopental, we recorded a significant intracellular [Ca2+] increase that was not from Ca2+ influx across the cell membrane, but from intracellular Ca2+ stores. The thiopental-induced [Ca2+]i rise in β-cells was sensitive to thapsigargin, a blocker of the endoplasmic reticulum Ca2+ pump, as well as to heparin (0.1 mg/ml) and 2-aminoethoxydiphenyl borate (2-APB; 100 μM), drugs that inhibit inositol 1,4,5-trisphosphate (IP3) binding to the IP3 receptor, and to U-73122, a phospholipase C inhibitor, but insensitive to ryanodine. Thapsigargin also diminished thiopental-induced insulin secretion. Thus, we conclude that thiopental-induced insulin secretion is mediated by activation of the intracellular IP3-sensitive Ca2+ store.
- Published
- 2011
17. In vivo Imaging of Superoxide Flashes in Skeletal Muscle
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Noah Weisleder, Wang Wang, Xiao-Li Tian, Ming Zheng, Heping Cheng, Jianjie Ma, Wanrui Zhang, Shujiang Shang, Xianhua Wang, Yanru Wang, Xiuqin Zhang, Jingsong Zhou, Min Chen, Kaitao Li, and Huaqiang Fang
- Subjects
chemistry.chemical_classification ,Yellow fluorescent protein ,Cell type ,Reactive oxygen species ,Cell signaling ,biology ,Superoxide ,Cell ,Biophysics ,Skeletal muscle ,Mitochondrion ,Cell biology ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Biochemistry ,medicine ,biology.protein - Abstract
Emerging evidence suggests that reactive oxygen species (ROS) constitute a class of signaling molecules that regulate diverse cell functions including metabolism, muscle contractility and apoptosis. Recently we have developed and characterized a highly sensitive and reversible superoxide-selective probe, a circularly permuted yellow fluorescent protein (cpYFP), and demonstrated quantal and transient superoxide-producing events (superoxide flashes) within single mitochondria across multiple cell types (Wang et al, Cell, 132, 279).To further understand the physiological significance of flash events, we generated the pan-tissue mt-cpYFP transgenic mice expressing cpYFP in the mitochondria of multiple tissues. In vivo imaging of superoxide signals in gastrocnemius of transgenic mouse under anesthesia revealed mitochondrial superoxide flashes with similar properties (Fig). Further, superoxide flashes were also visualized in isolated skeletal muscle fibers transfected in vivo by electroporation with mt-cpYFP. Our findings support that mitochondrial superoxide flash activity is a physiologically relevant phenomenon that may participate in diverse aspects of cell function and signaling.View Large Image | View Hi-Res Image | Download PowerPoint Slide
- Published
- 2009
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