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Prediction of cell-cell communication patierns of dorsal root ganglion cells:single-cell RNA sequencing data analysis
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作者 Yanna Lian Cheng Wu +1 位作者 Li Liu Xiangyao Li 《Neural Regeneration Research》 SCIE CAS CSCD 2024年第6期1367-1374,共8页
Dorsal root ganglion neurons transmit peripheral somatic information to the central nervous system,and dorsal root ganglion neuron excitability affects pain perception.Dorsal root ganglion stimulation is a new approac... Dorsal root ganglion neurons transmit peripheral somatic information to the central nervous system,and dorsal root ganglion neuron excitability affects pain perception.Dorsal root ganglion stimulation is a new approach for managing pain sensation.Knowledge of the cell-cell communication among dorsal root ganglion cells may help in the development of new pain and itch management strategies.Here,we used the single-cell RNA-sequencing(scRNA-seq)database to investigate intercellular communication networks among dorsal root ganglion cells.We collected scRNA-seq data from six samples from three studies,yielding data on a total of 17,766 cells.Based on genetic profiles,we identified satellite glial cells,Schwann cells,neurons,vascular endothelial cells,immune cells,fibroblasts,and vascular smooth muscle cells.Further analysis revealed that eight types of dorsal root ganglion neurons mediated proprioceptive,itch,touch,mechanical,heat,and cold sensations.Moreover,we predicted several distinct forms of intercellular communication among dorsal root ganglion cells,including cell-cell contact,secreted signals,extracellular matrix,and neurotransmitter-mediated signals.The data mining predicted that Mrgpra3-positive neurons robustly express the genes encoding the adenosine Adora2b(A2B)receptor and glial cell line-derived neurotrophic factor family receptor alpha 1(GFRα-1).Our immunohistochemistry results confirmed the coexpression of the A2B receptor and GFRα-1.Intrathecal injection of the A2B receptor antagonist PSB-603 effectively prevented histamine-induced scratching behaviour in a dose-dependent manner.Our results demonstrate the involvement of the A2B receptor in the modulation of itch sensation.Furthermore,our findings provide insight into dorsal root ganglion cell-cell communication patterns and mechanisms.Our results should contribute to the development of new strategies for the regulation of dorsal root ganglion excitability. 展开更多
关键词 cell-cell communications CellChat dorsal root ganglion scRNA-Seq database
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Transcriptional regulatory network during axonal regeneration of dorsal root ganglion neurons:laser-capture microdissection and deep sequencing 被引量:1
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作者 Li-Li Zhao Tao Zhang +2 位作者 Wei-Xiao Huang Ting-Ting Guo Xiao-Song Gu 《Neural Regeneration Research》 SCIE CAS CSCD 2023年第9期2056-2066,共11页
The key regulators and regeneration-associated genes involved in axonal regeneration of neurons after injury have not been clarified.In high-throughput sequencing,various factors influence the final sequencing results... The key regulators and regeneration-associated genes involved in axonal regeneration of neurons after injury have not been clarified.In high-throughput sequencing,various factors influence the final sequencing results,including the number and size of cells,the depth of sequencing,and the method of cell separation.There is still a lack of research on the detailed molecular expression profile during the regeneration of dorsal root ganglion neuron axon.In this study,we performed lase r-capture microdissection coupled with RNA sequencing on dorsal root ganglion neurons at 0,3,6,and 12 hours and 1,3,and 7 days after sciatic nerve crush in rats.We identified three stages after dorsal root ganglion injury:early(3-12 hours),pre-regeneration(1 day),and regeneration(3-7 days).Gene expression patterns and related function enrichment res ults showed that one module of genes was highly related to axonal regeneration.We verified the up-regulation of activating transcription factor 3(Atf3),Kruppel like factor 6(Klf6),AT-rich inte raction domain 5A(Arid5α),CAMP responsive element modulator(Crem),and FOS like 1,AP-1 transcription factor Subunit(Fosl1) in dorsal root ganglion neurons after injury.Suppressing these transcription factors(Crem,Arid5o,Fosl1 and Klf6) reduced axonal regrowth in vitro.As the hub transcription factor,Atf3 showed higher expression and activity at the preregeneration and regeneration stages.G protein-coupled estrogen receptor 1(Gper1),inte rleukin 12a(Il12α),estrogen receptor 1(ESR1),and interleukin 6(IL6) may be upstream factors that trigger the activation of Atf3 during the repair of axon injury in the early stage.Our study presents the detailed molecular expression profile during axonal regeneration of dorsal root ganglion neurons after peripheral nerve injury.These findings may provide reference for the clinical screening of molecular targets for the treatment of peripheral nerve injury. 展开更多
关键词 Arid5a ATF3 Crem dorsal root ganglion Fosl1 KLF6 laser-capture microdissection NEURON smart-seq2 gene expression profile transcription factor
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Regulatory effect of nerve growth factor on release of substance P in cultured dorsal root ganglion neurons of rat
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作者 杨向东 刘真 +3 位作者 刘花香 王丽红 马春红 李振中 《Neuroscience Bulletin》 SCIE CAS CSCD 2007年第4期215-220,共6页
Objective To investigate the regulatory effects of nerve growth factor (NGF) on basal and capsaicin-induced release of neuropeptide substance P (SP) in primary cultured embryonic rat dorsal root ganglion (DRG) n... Objective To investigate the regulatory effects of nerve growth factor (NGF) on basal and capsaicin-induced release of neuropeptide substance P (SP) in primary cultured embryonic rat dorsal root ganglion (DRG) neurons. Methods DRGs were dissected from 15-day-old embryonic Wistar rats. DRG neurons were dissociated and cultured, and then exposed to different concentrations of NGF (10 ng/mL, 30 ng/mL, or 100 ng/mL) for 72 h. The neurons cultured in media without NGF served as control. RT-PCR were used for detecting the mRNAs of SP and vanilloid receptor 1 (VR1) in the DRG neurons. The SP basal and capsaicin (100 nmol/L)-induced release in the culture were measured by radioimmunoassay (RIA). Results SP mRNA and VR1 mRNA expression increased in primary cultured DRG neurons in a dose-dependent manner of NGF. Both basal release and capsaicin-evoked release of SP increased in NGF-treated DRG neurons compared with in control group. The capsaicin-evoked release of SP also increased in a dose-dependent manner of NGF. Conclusion NGF may promote both basal release and capsaicin-evoked release of SP. NGF might increase the sensitivity of nociceptors by increasing the SP mRNA or VR1 mRNA. 展开更多
关键词 nerve growth factor dorsal root ganglion CAPSAICIN vanilloid receptor 1 substance P
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Identification of differentially expressed genes in dorsal root ganglion in early diabetic rats
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作者 朱清 顾锦华 +1 位作者 朱红艳 徐济良 《Neuroscience Bulletin》 SCIE CAS CSCD 2008年第4期219-224,共6页
Objective To screen and identify differentially expressed genes in the dorsal root ganglion (DRG) in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of ... Objective To screen and identify differentially expressed genes in the dorsal root ganglion (DRG) in early experimental diabetic rats. Methods Diabetic model rats were induced by single intraperitoneal injection of streptozotocin (STZ). At the second week after STZ injection, the sensory nerve conduction velocities (SNCV) of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction (DD-PCR) was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot. Results The SNCV in the diabetic model group [n = 9, (45.25±10.38) m/s] reduced obviously compared with the control group [n = 8, (60.10± 11.92) m/s] (P 〈 0.05). Seven distinct cDNA clones, one was up-regulated gene and the others were downregulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA (accession No., BC059140), which had not been reported to relate to diabetic neuropathy. Conclusion These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy. 展开更多
关键词 differential display polymerase chain reaction silver staining MRNA dorsal root ganglion DIABETES RAT
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Estrogen affects neuropathic pain through upregulating N-methyl-D-aspartate acid receptor 1 expression in the dorsal root ganglion of rats 被引量:8
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作者 Chao Deng Ya-juan Gu +1 位作者 Hong Zhang Jun Zhang 《Neural Regeneration Research》 SCIE CAS CSCD 2017年第3期464-469,共6页
Estrogen affects the generation and transmission of neuropathic pain,but the specific regulatory mechanism is still unclear.Activation of the N-methyl-D-aspartate acid receptor 1(NMDAR1) plays an important role in t... Estrogen affects the generation and transmission of neuropathic pain,but the specific regulatory mechanism is still unclear.Activation of the N-methyl-D-aspartate acid receptor 1(NMDAR1) plays an important role in the production and maintenance of hyperalgesia and allodynia.The present study was conducted to determine whether a relationship exists between estrogen and NMDAR1 in peripheral nerve pain.A chronic sciatic nerve constriction injury model of chronic neuropathic pain was established in rats.These rats were then subcutaneously injected with 17β-estradiol,the NMDAR1 antagonist D(-)-2-amino-5-phosphonopentanoic acid(AP-5),or both once daily for 15 days.Compared with injured drug na?ve rats,rats with chronic sciatic nerve injury that were administered estradiol showed a lower paw withdrawal mechanical threshold and a shorter paw withdrawal thermal latency,indicating increased sensitivity to mechanical and thermal pain.Estrogen administration was also associated with increased expression of NMDAR1 immunoreactivity(as assessed by immunohistochemistry) and protein(as determined by western blot assay) in spinal dorsal root ganglia.This 17β-estradiol-induced increase in NMDAR1 expression was blocked by co-administration with AP-5,whereas AP-5 alone did not affect NMDAR1 expression.These results suggest that 17β-estradiol administration significantly reduced mechanical and thermal pain thresholds in rats with chronic constriction of the sciatic nerve,and that the mechanism for this increased sensitivity may be related to the upregulation of NMDAR1 expression in dorsal root ganglia. 展开更多
关键词 nerve regeneration peripheral nerve injury ESTROGEN 17Β-ESTRADIOL N-rnethyl-D-aspartic acid receptor 1 pain sciatic nerve chronic constriction injury neuropathic pain D(-)-2-amino-5-phosphonopentanoic acid dorsal root ganglion spinal cord IMMUNOREACTIVITY western blot assay neural regeneration
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Optimization of micropatterned poly(lactic-coglycolic acid) films for enhancing dorsal root ganglion cell orientation and extension 被引量:5
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作者 Ching-Wen Li Brett Davis +3 位作者 Jill Shea Himanshu Sant Bruce Kent Gale Jayant Agarwal 《Neural Regeneration Research》 SCIE CAS CSCD 2018年第1期105-111,共7页
Nerve conduits have been a viable alternative to the ‘gold standard’ autograft for treating small peripheral nerve gap injuries. However, they often produce inadequate functional recovery outcomes and are ineffectiv... Nerve conduits have been a viable alternative to the ‘gold standard’ autograft for treating small peripheral nerve gap injuries. However, they often produce inadequate functional recovery outcomes and are ineffective in large gap injuries. Ridge/groove surface micropatterning has been shown to promote neural cell orientation and guide growth. However, optimization of the ratio of ridge/groove parameters to promote orientation and extension for dorsal root ganglion (DRG) cells on poly(lactic-co-glycolic acid) (PLGA) films has not been previously conducted. Photolithography and micro-molding were used to define various combinations of ridge/groove dimensions on PLGA films. The DRG cells obtained from chicken embryos were cultured on micropatterned PLGA films for cell orientation and migration evaluation.Biodegradation of the films occurred during the test period, however, this did not cause deformation or distortion of the micropatterns. Results from the DRG cell orientation test suggest that when the ridge/groove ratio equals 1 (ridge/groove width parameters are equal, i.e., 10 μm/10 μm (even)), the degree of alignment depends on the size of the ridges and grooves, when the ratio is smaller than 1 (groove controlled) the alignment increases as the ridge size decreases, and when the ratio is larger than 1 (ridge controlled), the alignment is reduced as the width of the grooves decreases. The migration rate and neurite extension of DRG neurons were greatest on 10 μm/10 μm and 30 μm/30 μm micropatterned PLGA films. Based on the data, the 10 μm/10 μm and 30 μm/30 μm micropatterned PLGA films are the optimized ridge/groove surface patterns for the construction of nerve repair devices. 展开更多
关键词 nerve regeneration nerve repair neural cell migration neural cell alignment MICROPATTERN dorsal root ganglion topological cues neural regeneration
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PKCε Mediates Substance P Inhibition of GABA_A Receptors-Mediated Current in Rat Dorsal Root Ganglion 被引量:4
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作者 李丽 赵磊 +4 位作者 王洋 马克涛 石文艳 王英姿 司军强 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2015年第1期1-9,共9页
The mechanism underlying the modulatory effect of substance P(SP) on GABA-activated response in rat dorsal root ganglion(DRG) neurons was investigated. In freshly dissociated rat DRG neurons, whole-cell patch-clam... The mechanism underlying the modulatory effect of substance P(SP) on GABA-activated response in rat dorsal root ganglion(DRG) neurons was investigated. In freshly dissociated rat DRG neurons, whole-cell patch-clamp technique was used to record GABA-activated current and sharp electrode intracellular recording technique was used to record GABA-induced membrane depolarization. Application of GABA(1–1000 μmol/L) induced an inward current in a concentration-dependent manner in 114 out of 127 DRG neurons(89.8 %) examined with whole-cell patch-clamp recordings. Bath application of GABA(1–1000 μmol/L) evoked a depolarizing response in 236 out of 257(91.8%) DRG neurons examined with intracellular recordings. Application of SP(0.001–1 μmol/L) suppressed the GABA-activated inward current and membrane depolarization. The inhibitory effects were concentration-dependent and could be blocked by the selective neurokinin 1(NK1) receptors antagonist spantide but not by L659187 and SR142801(1 μmol/L, n=7), selective antagonists of NK2 and NK3. The inhibitory effect of SP was significantly reduced by the calcium chelator BAPTA-AM, phospholipase C(PLC) inhibitor U73122, and PKC inhibitor chelerythrine, respectively. The PKA inhibitor H-89 did not affect the SP effect. Remarkably, the inhibitory effect of SP on GABA-activated current was nearly completely removed by a selective PKCε inhibitor epilon-V1-2 but not by safingol and LY333531, selective inhibitors of PKCα and PKCβ. Our results suggest that NK1 receptor mediates SP-induced inhibition of GABA-activated current and membrane depolarization by activating intracellular PLC-Ca2+-PKCε cascade. SP might regulate the excitability of peripheral nociceptors through inhibition of the "pre-synaptic inhibition" evoked by GABA, which may explain its role in pain and neurogenic inflammation. 展开更多
关键词 peripheral nervous system substance P GABAA receptor protein kinase C dorsal root ganglion
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Differential Expression of Alpha-Adrenoceptor Subtypes in Rat Dorsal Root Ganglion after Chronic Constriction Injury 被引量:3
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作者 成洪聚 马克涛 +3 位作者 李丽 赵磊 王洋 司军强 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2014年第3期322-329,共8页
Summary: mRNAs of alpha-adrenoceptor (α-AR) subtypes are found in neurons in dorsal root ganglion (DRG) and change after peripheral nerve injury. In this study, the distribution of α-AR subtype proteins was stu... Summary: mRNAs of alpha-adrenoceptor (α-AR) subtypes are found in neurons in dorsal root ganglion (DRG) and change after peripheral nerve injury. In this study, the distribution of α-AR subtype proteins was studied in L5 DRG of normal rats and rats with chronic constriction injury of sciatic nerve (CCI). Using immunofluorescence technique, it was found that α1A-, α1B-, and α2A-AR proteins were expressed in large, medium, and small size neurons in normal DRG, and significantly increased in all size neurons 14 days after CCI. α1D- and α2C-AR was also expressed in all size neurons in normal DRG. However, α1D-AR was significantly increased and α2C-AR was decreased in small size neurons 14 days post CCI. α2B-AR neurons were not detectable in normal and CCI DRG. Co-expression of α1A- and α2A-AR in the same neuron was observed in normal DRG and increased post CCI. Collectively, these results indicated that there is distinct distribution of α-AR subtypes in DRG neurons, and the distribution and levels of expression of α-AR subtypes change differently after CCI. The up-regulation of α-AR subtypes in DRG neurons may play an important role in the process of generating and transmitting neuropathic pain. 展开更多
关键词 alpha-adrenoceptor chronic constriction injury dorsal root ganglion neuropathic pain NORADRENALINE
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Silencing the enhancer of zeste homologue 2,Ezh2,represses axon regeneration of dorsal root ganglion neurons 被引量:3
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作者 Ting-Ting Guo Ying Zhao +4 位作者 Wei-Xiao Huang Tao Zhang Li-Li Zhao Xiao-Song Gu Song-Lin Zhou 《Neural Regeneration Research》 SCIE CAS CSCD 2022年第7期1518-1525,共8页
Recovery from injury to the peripheral nervous system is different from that of the central nervous system in that it can lead to gene reprogramming that can induce the expression of a series of regeneration-associate... Recovery from injury to the peripheral nervous system is different from that of the central nervous system in that it can lead to gene reprogramming that can induce the expression of a series of regeneration-associated genes.This eventually leads to axonal regeneration of injured neurons.Although some regeneration-related genes have been identified,the regulatory network underlying axon regeneration remains largely unknown.To explore the regulator of axon regeneration,we performed RNA sequencing of lumbar L4 and L5 dorsal root ganglion(DRG)neurons at different time points(0,3,6,12 hours,1,3 and 7 days)after rat sciatic nerve crush.The isolation of neurons was carried out by laser capture microscopy combined with NeuN immunofluorescence staining.We found 1228 differentially expressed genes in the injured sciatic nerve tissue.The hub genes within these differentially expressed genes include Atf3,Jun,Myc,Ngf,Fgf2,Ezh2,Gfap and Il6.We verified that the expression of the enhancer of zeste homologue 2 gene(Ezh2)was up-regulated in DRG neurons after injury,and this up-regulation differed between large-and small-sized dorsal root ganglion neurons.To investigate whether the up-regulation of Ezh2 impacts axonal regeneration,we silenced Ezh2 with siRNA in cultured DRG neurons and found that the growth of the newborn axons was repressed.In our investigation into the regulatory network of Ezh2 by interpretive phenomenal analysis,we found some regulators of Ezh2(including Erk,Il6 and Hif1a)and targets(including Atf3,Cdkn1a and Smad1).Our findings suggest that Ezh2,as a nerve regeneration-related gene,participates in the repair of the injured DRG neurons,and knocking down the Ezh2 in vitro inhibits the axonal growth of DRG neurons.All the experimental procedures approved by the Administration Committee of Experimental Animals of Jiangsu Province of China(approval No.S20191201-201)on March 21,2019. 展开更多
关键词 axon regeneration dorsal root ganglion neurons EZH2 IB4 laser capture microscopy NF160/200 quantitative reverse transcription-polymerase chain reaction sciatic nerve crush scRNA-seq siRNA
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Brain-derived neurotrophic factor expression in dorsal root ganglion neurons in response to reanastomosis of the distal stoma after nerve grafting 被引量:2
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作者 Wei Yu Jian Wang +2 位作者 Mingzhu Xu Hanjiao Qin Shusen Cui 《Neural Regeneration Research》 SCIE CAS CSCD 2012年第26期2012-2017,共6页
Studies have shown that retreatment of the distal stoma after nerve grafting can stimulate nerve regeneration. The present study attempted to verify the effects of reanastomosis of the distal stoma, after nerve grafti... Studies have shown that retreatment of the distal stoma after nerve grafting can stimulate nerve regeneration. The present study attempted to verify the effects of reanastomosis of the distal stoma, after nerve grafting, on nerve regeneration by assessing brain-derived neurotrophic factor expression in 2-month-old rats. Results showed that brain-derived neurotrophic factor expression in L2-4 dorsal root ganglia began to increase 3 days after autologous nerve grafting post sciatic nerve injury, peaked at 14 days, decreased at 28 days, and reached similar levels to the sham-surgery group at 56 days. Brain-derived neurotrophic factor expression in L2-4 dorsal root ganglia began to increase 3 days after reanastomosis of the distal stoma, 59 days after autologous nerve grafting post sciatic nerve injury, significantly increased at 63 days, peaked at 70 days, and gradually decreased thereafter, but remained higher compared with the sham-surgery group up to 112 days. The results of this study indicate that reanastomosis of the distal stoma after orthotopic nerve grafting stimulated brain-derived neurotrophic factor expression in L2.4 dorsal root ganglia. 展开更多
关键词 sciatic nerve orthotopic nerve grafting brain-derived neurotrophic factor dorsal root ganglion distalstoma reanastomosis peripheral nerve injury neural regeneration
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Characteristics of neural growth and cryopreservation of the dorsal root ganglion using three-dimensional collagen hydrogel culture versus conventional culture 被引量:2
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作者 Ze-Kai Cui Shen-Yang Li +6 位作者 Kai Liao Zhi-Jie Wang Yong-Long Guo Luo-Sheng Tang Shi-Bo Tang Jacey Hongjie Ma Jian-Su Chen 《Neural Regeneration Research》 SCIE CAS CSCD 2021年第9期1856-1864,共9页
In vertebrates, most somatosensory pathways begin with the activation of dorsal root ganglion(DRG) neurons. The development of an appropriate DRG culture method is a prerequisite for establishing in vitro peripheral n... In vertebrates, most somatosensory pathways begin with the activation of dorsal root ganglion(DRG) neurons. The development of an appropriate DRG culture method is a prerequisite for establishing in vitro peripheral nerve disease models and for screening therapeutic drugs. In this study, we compared the changes in morphology, molecular biology, and transcriptomics of chicken embryo DRG cultured on tissue culture plates(T-DRG) versus three-dimensional collagen hydrogels(C-DRG). Our results showed that after 7 days of culture, the transcriptomics of T-DRG and C-DRG were quite different. The upregulated genes in C-DRG were mainly related to neurogenesis, axon guidance, and synaptic plasticity, whereas the downregulated genes in C-DRG were mainly related to cell proliferation and cell division. In addition, the genes related to cycles/pathways such as the synaptic vesicle cycle, cyclic adenosine monophosphate signaling pathway, and calcium signaling pathway were activated, while those related to cell-cycle pathways were downregulated. Furthermore, neurogenesis-and myelination-related genes were highly expressed in C-DRG, while epithelial–mesenchymal transition-, apoptosis-, and cell division-related genes were suppressed. Morphological results indicated that the numbers of branches, junctions, and end-point voxels per C-DRG were significantly greater than those per T-DRG. Furthermore, cells were scattered in T-DRG and more concentrated in C-DRG, with a higher ratio of 5-ethynyl-2′-deoxyuridine(EdU)-positive cells in T-DRG compared with C-DRG. C-DRG also had higher S100 calcium-binding protein B(S100 B) and lower α-smooth muscle actin(α-SMA) expression than T-DRG, and contained fewer terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)-positive cells after 48 hours of serum starvation. After cryopreservation, C-DRG maintained more intact morphological characteristics, and had higher viability and less TUNEL-positive cells than T-DRG. Furthermore, newly formed nerve bundles were able to grow along the existing Schwann cells in C-DRG. These results suggest that C-DRG may be a promising in vitro culture model, with better nerve growth and anti-apoptotic ability, quiescent Schwann cells, and higher viability. Results from this study provide a reference for the construction, storage, and transportation of tissue-engineered nerves. The study was approved by the Ethics Committee of Aier School of Ophthalmology, Central South University, China(approval No. 2020-IRB16), on March 15, 2020. 展开更多
关键词 ANTI-APOPTOSIS collagen hydrogel CRYOPRESERVATION dorsal root ganglion NEUROGENESIS RNA-seq Schwann cell tissue engineering
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Effects of TRPA1 activation and inhibition on TRPA1 and CGRP expression in dorsal root ganglion neurons 被引量:2
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作者 Xiao-Lei Wang Li-Wei Cui +5 位作者 Zhen Liu Yue-Ming Gao Sheng Wang Hao Li Hu-Xiang Liu Ling-Jia Yu 《Neural Regeneration Research》 SCIE CAS CSCD 2019年第1期140-148,共9页
Transient receptor potential ankyrin 1 (TRPA1) is a key player in pain and neurogenic inflammation, and is localized in nociceptive primary sensory dorsal root ganglion (DRG) neurons. TRPA1 plays a major role in t... Transient receptor potential ankyrin 1 (TRPA1) is a key player in pain and neurogenic inflammation, and is localized in nociceptive primary sensory dorsal root ganglion (DRG) neurons. TRPA1 plays a major role in the transmission of nociceptive sensory signals. The generation of neurogenic inflammation appears to involve TRPAl-evoked release of calcitonin gene-related peptide (CGRP). However, it remains unknown whether TRPA1 or CGRP expression is affected by TRPA 1 activation. Thus, in this study, we examined TRPA 1 and CGRP expression in DRG neurons in vitro after treatment with the TRPA1 activator tbrmaldehyde or the TRPA1 blocker menthol. In addition, we examined the role of extracellular signal-regulated protein kinase 1/2 (ERK1/2) in this process. DRG neurons in culture were exposed to formaldehyde, menthol, the ERK1/2 inhibitor PD98059 + formaldehyde, or PD98059 + menthol. After treatment, real-time polymerase chain reaction, western blot assay and double immunofluorescence labeling were performed to evaluate TRPA1 and CGRP expression in DRG neurons. Formaldehyde elevated mRNA and protein levels of TRPA 1 and CGRP, as well as the proportion of TRPA1- and CGRP-positive neurons. In contrast, menthol reduced TRPA1 and CGRP expression. Furthermore, the effects of lbrmaldehyde, but not menthol, on CGRP expression were blocked by pretreatment with PD98059. PD98059 pretreatment did not affect TRPA1 expression in the presence of formaldehyde or menthol. 展开更多
关键词 nerve regeneration TRPA1 TRPV1 FORMALDEHYDE MENTHOL CGRP dorsal root ganglion NEURON neurogenic inflammation nociceptive signal ERK1/2 neural regeneration
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Effects of Intrathecally Administerd NaV1.8 Antisense Oligonucleotide on the Expression of Sodium Channel mRNA in Dorsal Root Ganglion 被引量:2
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作者 刘甬民 姚尚龙 +3 位作者 宋文阁 王月兰 刘东 曾涟 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2005年第6期696-699,共4页
Neuropathic pain has been hypothesized to be the result of aberrant expression and function of sodium channels at the site of injury. To investigate the effects of NaV1.8 antisense oligonucleotide on the expression of... Neuropathic pain has been hypothesized to be the result of aberrant expression and function of sodium channels at the site of injury. To investigate the effects of NaV1.8 antisense oligonucleotide on the expression of sodium channel mRNA in dorsal root ganglion (DRG) neurons in chronic neuropathic pain. 24 Sprague-Dawley rats weighing 200--260 g were anesthetized with the intraperitoneal injection of 300 mg· kg^-1 choral hydrate. The CCI model was made by loose ligation of sciatic nerve trunk by 4--0 chromic gut. The mechanical and thermal pain threshold were measured before operation and 1, 3, 5, 7, 9, 11, 13 days after operation. A PE-10 catheter was implanted in subarachnoid space at lumbar region. On the 7th postoperative day the animals were randomly divided into 4 groups. The drugs were injected intrathecally twice a day for 5 consecutive days in group 2--4. The animals were decapitated 14 days after the surgery. The L4--L6 DRG of the operated side was removed and crushed, and total RNA was extracted with Trizol reagent. The contralateral side was used as control. The change of NaV1.8 sodium channel transcripts was determined by RT-PCR. Pain threshold was significantly lowered after CCI as compared with that in control group and was elevated 3 days after antisense oligonucleotide injection. Sensory neuron specific TTX-R sodium channel NaV1.8 transcript was down-regulated after antisense oligonucleotide injection at the dosage of 45 μg as compared with that in CCI group (P〈0.01), and it was even greater at the dosage of 90 μg. The intrathecally injected NaV1.8 antisense oligonucleotide can reduce the mechanical allodynia and thermal hyperalgesia partially by downregulating the SNS transcript expression. 展开更多
关键词 tetrodotoxin-resistant sodium channel current neuropathic pain ANTISENSE dorsal root ganglion sensory neurons voltage sensing sodium channel type 1.8 (NaV1.8)
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Expression of three distinct families of calcium-activated chloride channel genes in the mouse dorsal root ganglion 被引量:2
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作者 Mohammed AL-JUMAILY Alexei KOZLENKOV +5 位作者 Ilana MECHALY Agnes FICHARD Valerie MATHA Frederique SCAMPS Jean VALMIER Patrick CARROLL 《Neuroscience Bulletin》 SCIE CAS CSCD 2007年第5期293-299,共7页
Objective A calcium-activated chloride current (ICl(Ca)) has been observed in medium-sized sensory neurons of the dorsal root ganglion (DRG). Axotomy of the sciatic nerve induces a similar current in the majorit... Objective A calcium-activated chloride current (ICl(Ca)) has been observed in medium-sized sensory neurons of the dorsal root ganglion (DRG). Axotomy of the sciatic nerve induces a similar current in the majority of medium and large diameter neurons. Our aim is to identify the molecule(s) underlying this current. Methods Using conventional and quantitative RT-PCR, we examined the expression in DRG of members of three families of genes, which have been shown to have latch) current inducing properties. Results We showed the detection of transcripts representing several members of these families, i.e. chloride channel calciumactivated (CLCA), Bestrophin and Tweety gene families in adult DRG, in the normal state and 3 d after sciatic nerve section, a model for peripheral nerve injury. Conclusion Our analysis revealed that that mBestl and Tweety2 appear as the best candidates to play a role in the injury-induced Icl(Ca) in DRG neurons. 展开更多
关键词 chloride channel calcium activated BESTROPHIN tweety chloride channel dorsal root ganglion
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srGAP3 promotes neurite outgrowth of dorsal root ganglion neurons by inactivating RAC1 被引量:2
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作者 Quan-Peng Zhang Hai-Ying Zhang +4 位作者 Xian-Fang Zhang Jiu-Hong Zhao Zhi-Jian Ma Dan Zhao Xi-Nan Yi 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2014年第8期630-638,共9页
Objective:To explore effect of srGAP3 promotes neurite outgrowth of dorsal root ganglion neurons.Methods:In this study,expression of Slit1 was observed predominantly in the glia.while expression of Robo2 and srCAP3 wa... Objective:To explore effect of srGAP3 promotes neurite outgrowth of dorsal root ganglion neurons.Methods:In this study,expression of Slit1 was observed predominantly in the glia.while expression of Robo2 and srCAP3 was detected in sensory neurons of postnatal rat cultured dorsal root ganglion(DRG).Furthermore,upregulation of srGAP3 following sciatic nerve transection was detected by immunohistochemistry and Western blotting.Results:It was observed that inhibition of nenrite outgrowth in cultured adult DRG neurons following treatment with anti-srGAP3 or anti-Robo2 was more effectively(1.5-fold higher) than that following treatment with an anti-BDNF positive control antibody.It demonstrated that srGAP3 interacted with Robo2 and Slit1 protein to decrease Rac1-CTP activity in cultured adult rat DRG neurons and the opposite effect on Rac1-GTP activity was detected by co-immunoprecipitation and immunoblotting analyses following treatment with anti-Robo2 or anti-srGAP3.These data demonstrated a role for srGAP3 in nenrite outgrowth ol DRG sensory neurons.Conclusions:Our observations suggest that srGAP3 promotes neurile outgrowth and filopodial growth cones by interacting with Robo2 to inactivate Rac1 in mammalian DRG neurons. 展开更多
关键词 RATS dorsal root ganglion Slit-Robo GTPase activating protein 3
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Low-frequency electrical stimulation improves neurite outgrowth of dorsal root ganglion neurons in vitro via upregulating Ca^(2+)-mediated brain-derived neurotrophic factor expression 被引量:1
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作者 Lidan Wan Rong Xia Wenlong Ding 《Neural Regeneration Research》 SCIE CAS CSCD 2010年第16期1256-1260,共5页
Short-term, low-frequency electrical stimulation of neural tissues significantly enhances axonal regeneration of peripheral nerves following injury. However, little is known about the mechanisms of electrical stimulat... Short-term, low-frequency electrical stimulation of neural tissues significantly enhances axonal regeneration of peripheral nerves following injury. However, little is known about the mechanisms of electrical stimulation to induce neurite outgrowth. In the present study, short-term, low-frequency electrical stimulation, using identical stimulation parameters of in vivo experiments, was administered to in vitro dorsal root ganglion (DRG) neurons. Enhanced neurite outgrowth, as well as synthesis and release of brain-derived neurotrophic factor (BDNF), were examined in electrical stimulation-treated DRG neuronal cultures. Because the effects of electrical stimulation on neuronal intracellular signaling molecules are less reported, classic calcium intracellular signals are directly or indirectly involved in electrical stimulation effects on neurons. Cultured DRG neurons were pretreated with the calcium channel blocker nifedipine, followed by electrical stimulation. Results suggested that electrical stimulation not only promoted in vitro neurite outgrowth, but also enhanced BDNF expression. However, nifedipine reduced electrical stimulation-enhanced neurite outgrowth and BDNF biosynthesis. These results suggest that the promoting effects of electrical stimulation on DRG neurite outgrowth could be associated with altered calcium influx, which is involved induction of neuronal BDNF expression and secretion. 展开更多
关键词 electrical stimulation dorsal root ganglion neurons neurite outgrowth brain-derived neurotrophic factor Ca2+ neural regeneration
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Expression of gamma-aminobutyric acid type A receptor α_2 subunit in the dorsal root ganglion of rats with sciatic nerve injury 被引量:1
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作者 Yu Lian Yang Wang +5 位作者 Ketao Ma Lei Zhao Zhongshuang Zhang Yuanyuan Shang Junqiang Si Li Li 《Neural Regeneration Research》 SCIE CAS CSCD 2012年第32期2492-2499,共8页
The γ-aminobutyric acid neurotransmitter in the spinal cord dorsal horn plays an important role in pain modulation through primary afferent-mediated presynaptic inhibition. The weakening of γ-aminobutyric acid-media... The γ-aminobutyric acid neurotransmitter in the spinal cord dorsal horn plays an important role in pain modulation through primary afferent-mediated presynaptic inhibition. The weakening of γ-aminobutyric acid-mediated presynaptic inhibition may be an important cause of neuropathic pain. γ-aminobutyric acid-mediated presynaptic inhibition is related to the current strength of γ-aminobutyric acid A receptor activation. In view of this, the whole-cell patch-clamp technique was used here to record the change in muscimol activated current of dorsal root ganglion neurons in a chronic constriction injury model. Results found that damage in rat dorsal root ganglion neurons following application of muscimol caused concentration-dependent activation of current, and compared with the sham group, its current strength and γ-aminobutyric acid A receptor protein expression decreased. Immunofluorescence revealed that γ-aminobutyric acid type A receptor α2 subunit protein expression decreased and was most obvious at 12 and 15 days after modeling. Our experimental findings confirmed that the y-aminobutyric acid type A receptor α2 subunit in the chronic constriction injury model rat dorsal root ganglion was downregulated, which may be one of the reasons for the reduction of injury in dorsal root ganglion neurons following muscimol-activated currents. 展开更多
关键词 γ-aminobutyric acid γ-aminobutyric acid type A receptor α2 subunit neuropathic pain dorsal root ganglion whole-cell patch clamp IMMUNOFLUORESCENCE primary afferent depolarization paw withdrawal latency MUSCIMOL
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Preemptive analgesic effects of low-dose ketamine on growth-associated protein expression in dorsal root ganglion of chronic constriction injury model rats 被引量:1
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作者 Shuyong Lin Chen Wang 《Neural Regeneration Research》 SCIE CAS CSCD 2008年第4期354-357,共4页
BACKGROUND: Ketamine is a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonists and plays an important role in the treatment of pain. OBJECTIVE: To analyze the preemptive analgesic effects of different d... BACKGROUND: Ketamine is a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonists and plays an important role in the treatment of pain. OBJECTIVE: To analyze the preemptive analgesic effects of different doses of ketamine on growth-associated protein-43 (GAP43) expression in dorsal root ganglion in a rat model of chronic sciatic nerve constricted injury, and to study the differences between high-dose and low-dose ketamine DESIGN: Randomized controlled animal study. SETTING: Medical College of Shantou University. MATERIALS: Thirty-five adult male Sprague Dawley rats were provided by the Experimental Animal Center of Guangzhou University of Traditional Chinese Medicine. Ketamine hydrochloride injection was provided by Hengrui Pharmaceutical Co., Ltd., Jiangsu. METHODS: This study was performed at the Immunological Laboratory, Medical College of Shantou University from September to December 2006. Model of chronic sciatic nerve constricted injury: after anesthesia, the right sciatic nerve was exposed and ligated l-cm distal to the ischiadic tuberosity with a No. 3-0 cat gut suture. Grouping and intervention: 35 rats were randomly divided into 4 groups: normal control group (n = 5), chronic constriction injury (CCI) group (n = 10), low-dose ketamine group (n = 10), and high-dose ketamine group (n = 10). Rats in the normal control group did not undergo any surgery or drug intervention. Rats in the CCI group received intraperitoneal injection of saline (1 mL), and their sciatic nerves were ligated after 10 minutes. Rats in the low-dose ketamine group underwent intraperitoneal injection of ketamine (25 mg/kg) 10 minutes prior to ligation of sciatic nerve; while, rats in the high-dose ketamine group were given intraperitoneal injection of ketamine (50 mg/kg) 10 minutes prior to ligation of sciatic nerve. On the third and the seventh days after surgery, dorsal root ganglion were resected from the sciatic nerve and cut into sections. MAIN OUTCOME MEASURES: GAP-43 expression in dorsal root ganglion was detected by immunohistochemistry and image analysis system, as well as semi-quantitative analysis. RESULTS: Thirty-five Sprague Dawley rats were included in the final analysis. Qualitative analysis: GAP-43 expression in the CCI group was higher than in the normal control group. Quantitative analysis: after three post-operative days, GAP-43 expression in the CCI group was significantly higher than in the normal control group (t = 22.919, 7.319, P 〈 0.05). GAP-43 expression in the low-dose and high-dose ketamine group was significantly lower than in the CCI group (t = 11.166, 26.474, P 〈 0.05). After seven postoperative days, GAP-43 expression in the low-dose and high-dose ketamine groups was significantly lower than in the CCI group (t = 2.382, 5.016, P 〈 0.05). CONCLUSION: Preoperative administration of ketamine inhibited the increased GAP-43 expression in dorsal root ganglion during neuropathic pain. 展开更多
关键词 growth-associated protein-43 neuropathic pain ketamlne sciatic nerve dorsal root ganglion
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In vitro neuroprotective effects of ciliary neurotrophic factor on dorsal root ganglion neurons with glutamate-induced neurotoxicity 被引量:6
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作者 Shu-yun Wen Ai-min Li +4 位作者 Kuan-qing Mi Rui-zheng Wang Hao Li Hua-xiang Liu Yi Xing 《Neural Regeneration Research》 SCIE CAS CSCD 2017年第10期1716-1723,共8页
Ciliary neurotrophic factor has neuroprotective effects mediated through signal transducer and Janus kinase(JAK) 2/activator of transcription 3(STAT3) and phosphatidylinositol 3-kinase(PI3 K)/Akt signaling pathw... Ciliary neurotrophic factor has neuroprotective effects mediated through signal transducer and Janus kinase(JAK) 2/activator of transcription 3(STAT3) and phosphatidylinositol 3-kinase(PI3 K)/Akt signaling pathways.Whether ciliary neurotrophic factor is neuroprotective for glutamate-induced excitotoxicity of dorsal root ganglion neurons is poorly understood.In the present study,the in vitro neuroprotective effects of ciliary neurotrophic factor against glutamate-induced excitotoxicity were determined in a primary culture of dorsal root ganglion neurons from Wistar rat embryos at embryonic day 15.Whether the JAK2/STAT3 and PI3 K/Akt signaling pathways were related to the protective effects of ciliary neurotrophic factor was also determined.Glutamate exposure inhibited neurite outgrowth,cell viability,and growth-associated protein 43 expression and promoted apoptotic neuronal cell death,all of which were reversed by the administration of exogenous ciliary neurotrophic factor.Additionally,preincubation with either JAK2 inhibitor AG490 or PI3 K inhibitor LY294002 blocked the neuroprotective effect of ciliary neurotrophic factor.These data indicate that the two pathways JAK2/STAT3 and PI3 K/Akt play major roles in mediating the in vitro neuroprotective effects of ciliary neurotrophic factor on dorsal root ganglion neurons with glutamate-induced neurotoxicity. 展开更多
关键词 nerve regeneration ciliary neurotrophic factor JAK2/STAT3 PI3K/Akt glutamate neuron excitotoxicity neuroprotection growth-associated protein 43 neurite outgrowth dorsal root ganglion neural regeneration
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The sensitivity of neurons with non-periodic activity to sympathetic stimulation in rat injured dorsal root ganglion 被引量:1
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作者 Hong-Jun YANG San-Jue HU +1 位作者 Pu-Lin GONG Jian-Hong DUAN 《Neuroscience Bulletin》 SCIE CAS CSCD 2006年第1期14-20,共7页
Objective The relationship between compressed dorsal root ganglion (DRG) neurons and firing pattern and sensitivity of neurons was studied in chronically the Hindmarsh-Rose (HR) neuronal model. Methods Spontane- o... Objective The relationship between compressed dorsal root ganglion (DRG) neurons and firing pattern and sensitivity of neurons was studied in chronically the Hindmarsh-Rose (HR) neuronal model. Methods Spontane- ous activities from single fibers of chronically compressed DRG neurons in rats were recorded, and divided into periodic and non-periodic firing patterns. The sensitivity of the two kinds of firing pattern neuron to sympathetic stimulation (SS) was compared. Result It was found that 27.3% of periodic firing neurons and 93.2% of non-periodic firing neurons responded to SS respectively ( periodic vs non-periodic, P 〈 0.01 ). The responses to SS with different stimulation time were greater non-periodic firing neurons than periodic firing neurons (P 〈 0.01 ). The non-periodic firing neurons obviously responded to SS. After the firing pattern of these neurons transformed to periodic firing pattern, their responses to SS disappeared or decreased obviously. The HR neuronal model exhibited a significantly greater response to perturbation in non-periodic (chaotic) firing pattern than in periodic firing pattern. Conelusion The non-periodic firing neurons with deterministic chaos are more sensitive to external stimuli than the periodic firing neurons. 展开更多
关键词 dorsal root ganglion Hindmarsh-Rose neuronal model spontaneous activity sympathetic stimulation sensitivity CHAOS
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