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Expression and effect of sodium-potassium-chloride cotransporter on dorsal root ganglion neurons in a rat model of chronic constriction injury

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Neural Regeneration Research 2020年第5期15卷 912-921页

作者： Chao-Yang Tan Yan-Ping Wang Yuan-Yuan Han Bi-Han Lu Wei Ji Li-Cang Zhu Yang Wang Wen-Yan Shi Li-Ya Shan Liang Zhang Ke-Tao Ma Li Li Jun-Qiang Si Department of Physiology College of MedicineShihezi UniversityShiheziXinjiang Uygur Autonomous RegionChina Department of Physiology Medical College of Jiaxing UniversityJiaxingZhejiang ProvinceChina Department of Health Karamay Army DivisionChinese People’s Liberation ArmyKaramayXinjiang Uygur Autonomous RegionChina Department of Nursing Medical College of Jiaxing UniversityJiaxingZhejiang ProvinceChina Department of Clinical Medicine Karamay College of Xinjiang Medical UniversityKaramayXinjiang Uygur Autonomous RegionChina Department of Neurosurgery First Affiliated HospitalCollege of MedicineShihezi UniversityShiheziXinjiang Uygur Autonomous RegionChina The key Laboratory of Xinjiang Endemic and Ethnic Diseases College of MedicineShihezi UniversityShiheziXinjiang Uygur Autonomous RegionChina Department of Physiology School of Basic Medical SciencesWuhan UniversityWuhanHubei ProvinceChina Department of Physiology School of Basic Medical SciencesHuazhong University of Science and TechnologyWuhanHubei ProvinceChina

Sodium-potassium-chloride cotransporter 1 (NKCC1) and potassium-chloride cotransporter 2 (KCC2) are associated with the transmission of peripheral pain.We investigated whether the increase of NKCC1 and KCC2 is associated with peripheral pain transmission in dorsal root ganglion neurons.To this aim,rats with persistent hyperalgesia were randomly divided into four groups.Rats in the control group received no treatment,and the rat sciatic nerve was only exposed in the sham group.Rats in the chronic constriction injury group were established into chronic constriction injury models by ligating sciatic nerve and rats were given bumetanide,an inhibitor of NKCC1,based on chronic constriction injury modeling in the chronic constriction injury + bumetanide group.In the experiment measuring thermal withdrawal latency,bumetanide (15 mg/kg) was intravenously administered.In the patch clamp experiment,bumetanide (10 μg/μL) and acutely isolated dorsal root ganglion neurons (on day 14) were incubated for 1 hour,or bumetanide (5 μg/μL) was intrathecally injected.The Hargreaves test was conducted to detect changes in thermal hyperalgesia in rats.We found that the thermal withdrawal latency of rats was significantly decreased on days 7,14,and 21 after model establishment.After intravenous injection of bumetanide,the reduction in thermal retraction latency caused by model establishment was significantly inhibited.Immunohistochemistry and western blot assay results revealed that the immune response and protein expression of NKCC1 in dorsal root ganglion neurons of the chronic constriction injury group increased significantly on days 7,14,and 21 after model establishment.No immune response or protein expression of KCC2 was observed in dorsal root ganglion neurons before and after model establishment.The Cl^– (chloride ion) fluorescent probe technique was used to evaluate the change of Cl^– concentration in dorsal root ganglion neurons of chronic constriction injury model rats.We found that the relative optical density of N-(ethoxycarb

关键词： bumetanide chronic constriction injury dorsal root ganglion dorsal root reflex hyperalgesia KCC2 nerve regeneration neuropathic pain NKCC1 primary afferent depolarization whole-cell patch clamp

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Ecological significance of dorsal polymorphism in a Batesian mimic snake

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Current Zoology 2018年第6期64卷 745-753页

作者： Xavier SANTOS Jairo S.AZOR Sergio CORTES Elisa RODRIGUEZ Jose LARIOS Juan M.PLEGUEZUELOS CIBIO/InBIO Centro de Investigacao em Biodiversidade e Recursos Geneticos da Universidade do PortoRua Padre Armando QuintasVairao 4485-661Portugal Departamento de Zoologia Facultad de CienciasUniversidad de GranadaGranada E-18071Spain Ayuntamiento de Motril Plaza de Espana 1Motril E-18600Spain

Batesian mimicry is the process in which harmless species adopt the appearance of a dangerous, aposematic species.In some prey species,both Batesian mimetic and non-Batesian morphs coexist,presupposing that both morphs have to be evolutionarily advantageous.The viperine snake, Natrix maura,exhibits a zigzag dorsal pattern and antipredatory behavior that mimics European vipers.This snake also has a striped dorsal pattern that coexists with the zigzag pattern.We have examined whether individuals belonging to different geographically structured clades were more likely to exhibit a certain dorsal pattern,and whether the zigzag pattern has a protective function by exposing artificial snakes to predation in natural environments,in addition to comparing antipredatory behavior between zigzag and striped snakes also in natural environments.Our results indicate that the striped pattern was not geographically structured,but habitat-dependent.Aerial predators less frequently attacked zigzag plasticine models than striped or unpatterned models.We detected a shift in antipredator behavior between the 2 morphs,as Batesian mimicking N.maura responded to an approaching potential predator by remaining immobile or fleeing at shorter distances than did striped ones.We conclude that Batesian mimics maintain the cryptic and aposematic value by resembling vipers,whereas in open habitats the non-Batesian mimic has altered its antipredator behavior to maintain its fitness.

关键词： antipredatory strategies aposematism Batesian mimicry crypsis dorsal pattern Natrix maura

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Hypoxia-ischemia in the immature rodent brain impairs serotonergic neuronal function in certain dorsal raphé nuclei

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Neural Regeneration Research 2020年第3期15卷 457-463页

作者： Hanna E.Reinebrant Julie A.Wixey Kathryn M.Buller UQ Centre for Clinical Research Faculty of Medicine The University of Queensland

Neonatal hypoxia-ischemia(HI)results in losses of serotonergic neurons in specific dorsal raphe nuclei.However,not all serotonergic raphe neurons are lost and it is therefore important to assess the function of remaining neurons in order to understand their potential to contribute to neurological disorders in the HI-affected neonate.The main objective of this study was to determine how serotonergic neurons,remaining in the dorsal raphe nuclei after neonatal HI,respond to an external stimulus(restraint stress).On postnatal day 3(P3),male rat pups were randomly allocated to one of the following groups:(i)control+no restraint(n=5),(ii)control+restraint(n=6),(iii)P3 HI+no restraint(n=5)or(iv)P3 HI+restraint(n=7).In the two HI groups,rat pups underwent surgery to ligate the common carotid artery and were then exposed to 6%O2 for 30 minutes.Six weeks after P3 HI,on P45,rats were subjected to restraint stress for 30 minutes.Using dual immunolabeling for Fos protein,a marker for neuronal activity,and serotonin(5-hydroxytrypamine;5-HT),numbers of Fos-positive 5-HT neurons were determined in five dorsal raphe nuclei.We found that restraint stress alone increased numbers of Fos-positive 5-HT neurons in all five dorsal raphe nuclei compared to control animals.However,following P3 HI,the number of stress-induced Fos-positive 5-HT neurons was decreased significantly in the dorsal raphe ventrolateral,interfascicular and ventral nuclei compared with control animals exposed to restraint stress.In contrast,numbers of stress-induced Fos-positive 5-HT neurons in the dorsal raphe dorsal and caudal nuclei were not affected by P3 HI.These data indicate that not only are dorsal raphe serotonergic neurons lost after neonatal HI,but also remaining dorsal raphe serotonergic neurons have reduced differential functional viability in response to an external stimulus.Procedures were approved by the University of Queensland Animal Ethics Committee(UQCCR958/08/NHMRC)on February 27,2009.

关键词： dorsal raphe nuclei Fos hypoxia-ischemia neonate newborn brain injury preterm restraint stress serotonin

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Gene expression changes in dorsal root ganglia following peripheral nerve injury: roles in inflammation,cell death and nociception

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Neural Regeneration Research 2019年第6期14卷 939-947页

作者： Sarah L.Martin Adam J.Reid Alexei Verkhratsky Valerio Magnaghi Alessandro Faroni Blond McIndoe Laboratories School of Biological SciencesUniversity of Manchester University Hospital of South Manchester Department of Plastic and BurnsManchester Faculty of Biology Medicine and HealthUniversity of Manchester Department of Pharmacological and Biomolecular Sciences Università degli Studi di Milano

Subsequent to a peripheral nerve injury, there are changes in gene expression within the dorsal root ganglia in response to the damage. This review selects factors which are well-known to be vital for inflammation, cell death and nociception, and highlights how alterations in their gene expression within the dorsal root ganglia can affect functional recovery. The majority of studies used polymerase chain reaction within animal models to analyse the dynamic changes following peripheral nerve injuries. This review aims to highlight the factors at the gene expression level that impede functional recovery and are hence are potential targets for therapeutic approaches. Where possible the experimental model, specific time-points and cellular location of expression levels are reported.

关键词： Gene expression polymerase chain reaction dorsal root ganglia inflammation nociception cell death peripheral nerve injury Schwann cells satellite glial cells nerve regeneration

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Altered Local Field Potential Relationship Between the Parafascicular Thalamic Nucleus and dorsal Striatum in Hemiparkinsonian Rats

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Neuroscience Bulletin 2019年第2期35卷 315-324页

作者： Haiyan Zhang Jing Yang Xuenan Wang Xiaomeng Yao Hongyu Han Yunfeng Gao Hongli Chang Tianyu Xiang Shuang Sun Yanan Wang Xiusong Wang Min Wang Key Laboratory of Animal Resistance Biology of Shandong Province College of Life Science Shandong Normal University School of Physics and Electronics Shandong Normal University

The thalamostriatal pathway is implicated in Parkinson's disease(PD); however, PD-related changes in the relationship between oscillatory activity in the centromedian-parafascicular complex(CM/Pf, or the Pf in rodents) and the dorsal striatum(DS) remain unclear.Therefore, we simultaneously recorded local field potentials(LFPs) in both the Pf and DS of hemiparkinsonian and control rats during epochs of rest or treadmill walking. The dopamine-lesioned rats showed increased LFP power in the beta band(12 Hz–35 Hz) in the Pf and DS during both epochs, but decreased LFP power in the delta(0.5 Hz–3 Hz) band in the Pf during rest epochs and in the DS during both epochs, compared to control rats. In addition,exaggerated low gamma(35 Hz–70 Hz) oscillations after dopamine loss were restricted to the Pf regardless of the behavioral state. Furthermore, enhanced synchronization of LFP oscillations was found between the Pf and DS after the dopamine lesion. Significant increases occurred in the mean coherence in both theta(3 Hz–7 Hz) and beta bands,and a significant increase was also noted in the phase coherence in the beta band between the Pf and DS during rest epochs. During the treadmill walking epochs, significant increases were found in both the alpha(7 Hz–12 Hz)and beta bands for two coherence measures. Collectively,dramatic changes in the relative LFP power and coherence in the thalamostriatal pathway may underlie the dysfunction of the basal ganglia-thalamocortical network circuits in PD, contributing to some of the motor and non-motor symptoms of the disease.

关键词： Parkinson’s disease Parafascicular thalamic nucleus dorsal striatum Local field potential Synchronization

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Decreased miR-325-5p Contributes to Visceral Hypersensitivity Through Post-transcriptional Upregulation of CCL2 in Rat dorsal Root Ganglia

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Neuroscience Bulletin 2019年第5期35卷 791-801页

作者： Rui Wu Ping-An Zhang Xuelian Liu Yuan Zhou Meijie Xu Xinghong Jiang Jun Yan Guang-Yin Xu Center for Translational Medicine Affiliated Zhangjiagang Hospital of Soochow University Zhangjiagang 215600 China Department of Physiology and Neurobiology Institute of Neuroscience Soochow University Suzhou 215123 China The Second Affiliated of Hospital Soochow University Suzhou 215004 China

Chronic visceral hypersensitivity is an important type of chronic pain with unknown etiology and pathophysiology. Recent studies have shown that epigenetic regulation plays an important role in the development of chronic pain conditions. However, the role of mi RNA-325-5 p in chronic visceral pain remains unknown. The present study was designed to determine the roles and mechanism of mi RNA-325-5 p in a rat model of chronic visceral pain.This model was induced by neonatal colonic inflammation(NCI). In adulthood, NCI led to a significant reduction in the expression of mi RNA-325-5 p in colon-related dorsal root ganglia(DRGs), starting to decrease at the age of4 weeks and being maintained to 8 weeks. Intrathecal administration of mi RNA-325-5 p agomir significantly enhanced the colorectal distention(CRD) threshold in a time-dependent manner. NCI also markedly increased the expression of CCL2(C-C motif chemokine ligand 2) in colon-related DRGs at the m RNA and protein levels relative to age-matched control rats. The expression of CXCL12, IL33, SFRS7, and LGI1 was not significantlyaltered in NCI rats. CCL2 was co-expressed in Neu Npositive DRG neurons but not in glutamine synthetasepositive glial cells. Furthermore, CCL2 was mainly expressed in isolectin B4-binding-and calcitonin generelated peptide-positive DRG neurons but in few NF-200-positive cells. More importantly, CCL2 was expressed in mi R-325-5 p-positive DRG neurons. Intrathecal injection of mi RNA-325-5 p agomir remarkably reduced the upregulation of CCL2 in NCI rats. Administration of Bindarit, an inhibitor of CCL2, markedly raised the CRD threshold in NCI rats in a dose-and time-dependent manner. These data suggest that NCI suppresses mi RNA-325-5 p expression and enhances CCL2 expression, thus contributing to visceral hypersensitivity in adult rats.

关键词： Visceral pain dorsal root ganglia miRNA-325-5p CCL2 Epigenetic regulation

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A novel primary culture method for high-purity satellite glial cells derived from rat dorsal root ganglion

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Neural Regeneration Research 2019年第2期14卷 339-345页

作者： Xian-Bin Wang Wei Ma Tao Luo Jin-Wei Yang Xiang-Peng Wang Yun-Fei Dai Jian-Hui Guo Li-Yan Li Institute of Neuroscience Kunming Medical University Medical Faculty Panzhihua University Second Department of General Surgery First People's Hospital of Yunnan Province

Satellite glial cells surround neurons within dorsal root ganglia. Previous studies have focused on single-cell suspensions of cultured neurons derived from rat dorsal root ganglia. At present, the primary culture method for satellite glial cells derived from rat dorsal root ganglia requires no digestion skill. Hence, the aim of the present study was to establish a novel primary culture method for satellite glial cells derived from dorsal root ganglia. Neonatal rat spine was collected and an incision made to expose the transverse protrusion and remove dorsal root ganglia. dorsal root ganglia were freed from nerve fibers, connective tissue, and capsule membranes, then rinsed and transferred to 6-well plates, and cultured in a humidified 5% CO_2 incubator at 37°C. After 3 days in culture, some cells had migrated from dorsal root ganglia. After subculture, cells were identified by immunofluorescence labeling for three satellite glial cell-specific markers: glutamine synthetase, glial fibrillary acidic protein, and S100β. Cultured cells expressed glutamine synthetase, glial fibrillary acidic protein, and S100β, suggesting they are satellite glial cells with a purity of > 95%. Thus, we have successfully established a novel primary culture method for obtaining high-purity satellite glial cells from rat dorsal root ganglia without digestion.

关键词： nerve regeneration cell culture dorsal root ganglia immunofluorescence identification satellite glial cells neural regeneration

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The Functional Linkage Among the "ZSL"-Spinal dorsal Horn-SN

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Science China Chemistry 1993年第10期36卷 1198-1206页

作者：孟卓吕国蔚 Department of Neurobiology Capital Institute of Medicine Beijing 100054 PRC Institute of Acupuncture and MoxibustionChina Academy of Traditional Chinese MedicineBeijing 100700PRC Department of Neurobiology Capital Institute of Medicine Beijing 100054 PRC

Electrical stimulation of the Zusanli point (ZSL) and solitary tract nucleus (SN) as well as microelectrode recording from the laminae Ⅲ-Ⅴ of the lumbar spinal dorsal horn have been used on the pentobarbital anesthetized rats, finding and identifying 57 spinal neurons responding to the stimulations of both ZSL and SN. Among them, 34 responded antidromically to SN; the others responded orthodromically to SN. Among them, the lowthreshold mechano-receptive(LTM) neurons and wide-dynamic-range (WDR) neurons were 50 percent respectively. The results indicate that (ⅰ) a single spinal dorsal horn neuron receives somatic afferent input and then conveys it to the visceral sensory nucleus-SN; (ⅱ) some spinal dorsal horn neurons receive, in turn, innervation from the SN; (ⅲ) the convergence and integration between somatic and visceral sensory inputs might occur in the spinal dorsal horn neurons and/or SN.

关键词： spinal dorsal horn Zusanli point solitary tract nucleus antidromic responses or thodromic responses.

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THE PROJECTION LINKAGE BETWEEN THE SPINAL dorsal HORN NEURONS AND BOTH THE SOLITARY TRACT AND dorsal COLUMN NUCLEI

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Science China Chemistry 1991年第2期34卷 171-183页

作者：吕国蔚孟卓罗蕾山上芳雄 Department of Neurobiology Capital Institute of Medicine Visiting fellow from the Chinese Academy of Tradiational Medicine Visiting fellow from Showa University

Electrical stimulation of the solitary tract nucleus (SN) and dorsal column nuclei (DCN)as well as microelectrode recording from the lumbal spinal dorsal horn have been used tofind and identify the axonal projection of and the afferent innervation on the spinal neuronsof pentobarbital- anesthetized rats. A total of 92 neurons was recorded and identified mainly in laminae Ⅲ-Ⅴ of the lumbarspinal dorsal horn. Of them, 38 neurons were activated antidromically from stimulation ofboth the SN and DCN. The other 54 neurons responded synaptically to both the SN andDCN stimulations. The initial antidromic responses of 8 neurons in the first group werefollowed by one or more responses synaptically driven from the SN and/or DCN stimulation.Conduction velocities were in the range of A delta fibers, but faster in the antidromicresponses and slower in the synaptic responses. These results indicate that (i) some spinal neurons issue branched axons of larger-sizedA delta fibers and double project to both the SN and

关键词： spinal dorsal horn solitary tract nucleus dorsal column nuclei antidromic responses synaptic responses

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Selective dorsal rhizotomy in cerebral palsy spasticity-a newly established operative technique in Slovenia: A case report and review of literature

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World Journal of Clinical Cases 2019年第10期7卷 1133-1141页

作者： Tomaz Velnar Peter Spazzapan Zoran Rodi Natasa Kos Roman Bosnjak Department of Neurosurgery University Medical Centre LjubljanaLjubljana 1000Slovenia AMEU-ECM Maribor Ljubljana 1000Slovenia Department of Neurophysiology University Medical Centre LjubljanaLjubljana 1000Slovenia Medical Rehabilitation Unit University Medical Centre LjubljanaLjubljana 1000Slovenia

BACKGROUND Spasticity affects a large number of children,mainly in the setting of cerebral palsy,however,only a few paediatric neurosurgeons deal with this problem.This is mainly due to the fact that until 1979,when Fasano has published the first series of selective dorsal rhizotomy(SDR),neurosurgeons were able to provide such children only a modest help.The therapy of spasticity has made a great progress since then.Today,peroral drugs,intramuscular and intrathecal medicines are available,that may limit the effects of the disease.In addition,surgical treatment is gaining importance,appearing in the form of deep brain stimulation,peripheral nerve procedures and SDR.All these options offer the affected children good opportunities of improving the quality of life.CASE SUMMARY A 15-year old boy is presented that was surgically treated for spasticity as a result of cerebral palsy.Laminotomy at L1 level was performed and L1 to S1 nerve roots were isolated and divided in smaller fascicles.Then,the SDR was made.CONCLUSION We describe a patient report and surgical technique of SDR that was performed in Slovenia for the first time.

关键词： Cerebral palsy Spasticity Selective dorsal rhizotomy Operation Case report

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