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Science China(Life Sciences) 2013年第8期56卷 818-822页

作者： ZHANG JiQian LI Xiang LIANG LiSi HUANG ShouFang ZHANG HengGui College of Physics and Electronic Information Anhui Normal University Biological Physics Group School of Physics & Astronomy The University of Manchester

Loss of function and gain of function mutations of the sodium channel were investigated using an intact two-dimensional rabbit sinoatrial node (SAN) and atrial cell model. The effects of three external stimuli (acetylcholine secretion by the vagal nerve, acid-base concentration, and tissue temperature) on cardiac pacemaker function and conduction were studied. Our results show that these two groups of mutations have different effects on pacemaker function and conduction. Furthermore, we found that the negative effects of these mutations could be altered by external stimuli. The bradycardic effects of mutations were magnified by an increase in acetylcholine level. Changes in acid-base concentration and tissue temperature increased the ability of the SAN to recover its pacemaker function. The results of this study increase our understanding of sodium channel disorders, and help to advance research on the treatment of these conditions.

关键词： gene mutation computer simulation sodium channel abnormal pacemaker 2D rabbit sinoatrial node cell model

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Two novel sodium channel mutations associated with resistance to indoxacarb and metaflumizone in the diamondback moth, Plutella xylostella

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Insect Science 2016年第1期23卷 50-58页

作者： Xing-Liang Wang Wen Su Jian-Heng Zhang Yi-Hua Yang Ke Dong Yi-Dong Wu College ofPlantProtection NanjingAgricultural University Nanjing China Department of Entomology Genetics and Neuroscience Programs Michigan State University East Lansing MI USA

Indoxacarb and metaflumizone belong to a relatively new class of sodium channel blocker insecticides （SCBIs）. Due to intensive use of indoxacarb, field-evolved indoxacarb resistance has been reported in several lepidopteran pests, including the dia- mondback moth Plutella xylostella, a serious pest of cruciferous crops. In particular, the BY 12 population of P. xylostella, collected from Baiyun, Guangdong Province of China in 2012, was 750-fold more resistant to indoxacarb and 70-fold more resistant to metaflu- mizone compared with the susceptible Roth strain. Comparison of complementary DNA sequences encoding the sodium channel genes of Roth and BY12 revealed two point mu- tations （F1845Y and V1848I） in the sixth segment of domain IV of the PxNav protein in the BY population. Both mutations are located within a highly conserved sequence region that is predicted to be involved in the binding sites of local anesthetics and SCBIs based on mammalian sodium channels. A significant correlation was observed among 10 field-collected populations between the mutant allele （Y 1845 or I 1848） frequencies （1.7% to 52.5%） and resistance levels to both indoxacarb （34- to 870-fold） and metaflumizone （1- to 70-fold）. The two mutations were never found to co-exist in the same allele of PxNav, suggesting that they arose independently. This is the first time that sodium channel muta- tions have been associated with high levels of resistance to SCBIs. F 1845Y and V 1848I are molecular markers for resistance monitoring in the diamondback moth and possibly other insect pest species.

关键词： indoxacarb metaflumizone mutation Plutellaxylostella resistance sodium channel

Equivalent intensity but differential dominance of sodium channel blocker insecticide resistance conferred by F1845Y and V1848l mutations of the voltage-gated sodium channel in Plutella xylostella

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Insect Science 2023年第1期30卷 125-134页

作者： Xingliang Wang Jianheng Zhang Yihua Yang Yidong Wu Key Laboratory of Integrated Pest Management on Crops in East China(MARA) College of Plant ProtectionNanjing Agricultural UniversityNanjingChina

Two point mutations(F1845Y and V1848I)in the voltage-gated sodium channel gene of Plutella xylostella are involved in the target-site resistance to sodium channel blocker insecticides(SCBIs).The contribution of the individual mutations to the SCBI resistance and the associated inheritance modes is as yet *** 2 rounds of single-pair crossing and marker-assisted selection,2 P xylostella strains(1845Y and 18481)bearing homozygous F1845Y or V18481 mutant alleles were successfully established from a field-collected population,and the contribution of each mutation to SCBI resistance,as well as associated inheritance patterns,was *** compared with the susceptible SZPS strain,each of the mutations individually conferred equally high-level resistance to indoxacarb(378 and 313 fold)and metaflumizone(734 and 674 fold),***,dominance levels of resistance to SCBIs were significantly different between the 2 resistant *** of the 1845Y strain to indoxacarb and metaflumizone was inherited as an autosomal and incompletely dominant trait(D values ranged from 0.43 to 0.76).In contrast,that of the 1848I strain followed an autosomal but incompletely recessive to semidominant mode(D values:-0.24 to 0.09).Our findings enriched the current understanding of inheritance and mechanisms of SCBI resistance in ***,and will help develop resistance management programs for P:xylostella and othereconomic pests.

关键词： diamondback moth indoxacarb inheritance metaflumizone resistance sodium channel

Effect of orphanin FQ and morphine on sodium channel current in somatosensory area of rat cerebral cortex

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Neural Regeneration Research 2007年第6期2卷 321-325页

作者： Lei Yang Yurong Li Shuwei Jia Yunhong Zhang Lanwei Cui Lihui Qu Department of Physiology Harbin Medical University Harbin 150086 Heilongjiang Province China

BACKGROUND： Some experiments have demonstrated that injecting orphanin FQ （OFQ） into lateral ventricle, which can obviously decrease the pain threshold. It is indicated that OFQ is an anti-opiate substance. However, whether OFQ has effects on sensory neuron ion channel in cerebral cortex needs to be further studied. OBJECTIVE： To investigate the effects of OFQ, morphine or their combination on sodium channel current of somatosensory neurons in rat cerebral cortex. DESIGN： Repeated measurement trial. SETTING： Department of Physiology, Harbin Medical University. MATERIALS： Fifty healthy Wistar rats, aged 12 - 16 days, of either gender, were provided by the Experimental Animal Center, Second Hospital Affiliated to Harbin Medical University. OFQ was purchased from Sigma-Aldrich Company, and morphine was provided by the Shenyang First Pharmaceutical Factory. PC2C patch clamp amplifier and LabmasterTLlwere purchased from Yibo Life Science Instrument Co.,Ltd. of Huazhong University of Science and Techgnology. METHODS： This experiment was carried out in the Department of Physiology （provincial laboratory）, Harbin Medical University between January 2005 and May 2006. Cortical neurons were acutely isolated from rats, and prepared into cell suspension following culture. ①sodium channel current of somatosensory neurons in rat cerebral cortex was recorded before and after administration by whole-cell Patch clamp technique after 50 nmol/L OFQ being added to extracellular fluid. ②The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex was recorded before and after administration by the same method after 20 μmol/L morphine being added to extracellular fluid, and then the change of sodium channel current was recorded after 50 nmol/L OFQ being added. MAIN OUTCOME MEASURES： The amplitude of sodium channel current of somatosensory neurons in rat cerebral cortex following the administration of OFQ, morphine separately or their combination.. RESULTS： ①The

关键词： orphnin morphine patch clamp sodium channel

Cloning of Partial sodium channel Gene From Strains of Fenvalerate-Resistant and Susceptible Cotton Aphid(Aphis gossypii Glover)

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Agricultural Sciences in China 2003年第8期2卷 881-885页

作者： SUN Lu-juan, GAO Xi-wu and ZHENG Bing-zong(Department of Entomology , China Agricultural University, Beijing 100094 , P.R.China) Department of Entomology China Agricultural University Beijing 100094 P.R.China

The strain of fenvalerate-resistant cotton aphids was selected using fenvalerate insecticide in the laboratory, the resistance factor of the strain was 199.54. Three degenerate primers were designed and used to perform PCR amplification. A cDNA encoding partial sodium channel gene was cloned from the fenvalerate-resistant and -susceptible strains. There were two nucleotide acid differences between fenvalerate-resistant strain and -susceptible strain, resulting in an amino acid mutation, Met→Leu. It is predicted that the mutation is related to the cotton aphid resistance to fenvalerate.

关键词： Cotton aphid Fenvalerate resistance sodium channel

Expression of hNav1.8 sodium channel protein in affected nerves of patients with trigeminal neuralgia

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Journal of Medical Colleges of PLA(China) 2004年第5期19卷 307-310页

作者：朱凌兰姜晓钟赵云富李玉莉何金 Department of Stomatology Changzheng Hospital Second Military Medical UniversityShanghai 200003 China Department of Stomatology Changzheng Hospital Second Military Medical UniversityShanghai 200003China Department of Pathology Changzheng Hospital Second Military Medical UniversityShanghai 200003China

Objective: To explore the pathogenesis of trigeminal neuralgia (TN) and to provide a new target for the drug treatment of TN by studying the expression of tetrodotoxin-resistant hNav1. 8 sodium channel protein in affected nerves of patients with TN. Methods: Twelve affected inferior alveolar nerves were obtained from patients with idiopathic TN, to whom the drug therapy was not effective. As negative control, one nonnal inferior alveolar nerve was obtained from patients who accepted the combined radical neck dissection with glossectomy and mandibulectomy. One muscle sample was obtained as normal control. One dorsal root ganglion from rat was as positive control. These tissues and prepared hNav1. 8 antibody were conducted immunohistochemistry response. Results: hNav1.8 channel protein was expresses in all the 12 specimens of the affected nerves of patients with TN, but not in the muscle sample and the normal inferior alveolar nerve. Conclusion: The abnormal expression of hNav1. 8 channel protein in the affected nerves of patients with TN may play an important role in the pathogenesis of TN.

关键词： tetrodotoxin sodium channel trigeminal neuralgia dorsal root ganglion trigeminal ganglion

Identifying the promoter region of the human brain sodium channel subtype Ⅰ gene（SCN1A） and subtype Ⅲ gene（SCN3A）

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Identifying the promoter region of the human brain sodium ch...

Decreased expression of sodium channel in the AIS of pyramidal cells in multiple sclerosis model mice

中国神经科学学会第四次会员代表大会暨第七届全国学术会议

作者： Yue-Sheng LONG,Wei-Ping LIAO~*,Qi-Hua ZHAO,Yi-Wu SHI,Wei-Yi DENG,Tao SU,Wei-Wen SUN,Mei-Mei GAO, Mei-Juan YU Institute of Neuroscience,Second Affiliated Hospital,Guangzhou Medical College,Guangzhou 510260,China

＜正＞Objective Voltage-gated sodium channels play critical role in the electrical excitability of the *** the known eleven sodium channel genes,four are broadly expressed throughout the central nervous system（CNS）:SCN1A,SCN2A, SCN3A,and SCN8A,which are also developmentally,regionally,and cellularly regulated in *** transcription of these genes may induce neurological *** the other hand,pathological insults can lead to alterations in sodium channel expression and ***,the transcriptional regulation of the sodium channel genes and its relationship with neurological disorders are poorly *** the CNS-specific sodium channels,the promoter regions and 5’ untranslated regions（UTRs） of human SCN2A and SCN8A have been described before,but the promoter regions and 5’ UTRs of human SCN1A and SCN3A have not been *** To identify the transcription start sites for SCN1A and SCN3A,5’-rapid amplification of cDNA ends（5’-RACE） was carried out on total RNA from human brain sections including hippocampus and cortex,and the reverse primer was located in the first coding exons of SCN1A and *** fragments were sequenced by cloning into pGEM-T *** blast analyses of the 5’-RACE products with human genomic sequence were performed using BLAST software from NCBI *** a 3-kb genomic fragment containing transcription initiator,untranslated exons and putative enhancers was cloned into the pGL4.10-Basic expression vector upstream of the firefly luciferase cDNA and the recombinant constructs were transformed into cultured neuroblast cells and HEK-293 *** promoter activities were assessed by Luciferase *** A 280-bp band for SCN1A and a 450-bp band for SCN3A were *** SCNIA,three untranslated exons（86 bp,92 bp and 49 bp,respectively） were dispersed in 5’ flanking region and the promoter was located 75 kb upstream of the first coding exon. For SCN3A,three untranslated exon（228 bp,197

关键词： promoter sodium channel 5’-RACE transcription element

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Decreased expression of sodium channel in the AIS of pyramid...

Cloning and characterization of the transcription regulatory region of human voltage-gated sodium channel SCN3A Gene

中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Xue-Mei CHEN,You-Sheng SHU~* Institute of Neuroscience,Shanghai Institutes for Biological Sciences,Chinese Academy Sciences,Shanghai,200031 China

Objective Multiple sclerosis(MS) is the most common neurological cause of disability in young adults. Specific Na channel isoforms have been shown to play important roles in the pathophysiology of *** this study,we explored the changes of Na channel expression at the axon initial segment(AIS) in demyelinated pyramidal cells,and tried to link these changes with the pathophysiological course of *** Two-month old C57BL/6 mice were fed with cuprizone-enriched food for 5 to 8 weeks to induce *** used MBP(myelin basic protein) staining to evaluate the success of the *** then performed immunostaining for Pan-Nav(total Na channels),Nav1.6 and AnkG (channel-association protein).Results In comparison with the control group,the relative density of both Pan-Nav and Nav 1.6 decreased at the AIS of pyramidal cells in MS *** Demyelination caused a reduction in channel density of Na channels at the AIS of pyramidal cells.

关键词： multiple sclerosis detnyelination sodium channel

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Cloning and characterization of the transcription regulatory...

Venus Flytrap HKT1-Type channel Provides for Prey sodium Uptake into Carnivorous Plant Without Conflicting with Electrical Excitability

中国神经科学学会第四次会员代表大会暨第八届全国学术会议

作者： Yue-Sheng LONG,Jia-Ming QIN,Xun-Sha SUN,Tao ZENG,Qi-Hua ZHAO,Wei-Ping LIAO~* Institute of Neuroscience and the Second Affiliated Hospital of Guangzhou Medical College Key Laboratory of Neurogenetics and channelopathies of Guangdong Province and the Ministry of Education of China,Guangzhou 510260,China

＜正＞Voltage-gated sodium channels are responsible for the electrical excitability of the *** sodium channel genes are expressed broadly throughout the central nervous system（CNS）:SCN1 A,SCN2A,and *** in SCN1A and SCN2A are associated with several neurology disorders:epilepsy,familial hemiplegic migraine,and familial autism,and one mutation in SCN3A had been reported in a patient with cryptogenic pediatric partial *** in SCN3A transcript was found in animal seizure models,epileptic patients and peripheral nerve injury patients, which is believed to play a pivotal role in these *** date,the transcriptional mechanism of SCN3A is poorly *** identify the transcription start sites（TSS） for SCN3A,5’-rapid amplification of cDNA ends（5’-RACE） was carried out on human brain total RNA（Clontech） and the RACE products were sequenced by cloning into pGEM-T *** blast analysis of the 5’-RACE products with human genomic sequence was performed using BLAST software from NCBI *** characterize the functional promoter region and transcriptional regulatory elements,a 1.1- kb genomic fragment,5’deletion fragments and element-deletion fragments were cloned into the pGL4.10-basic expression vector upstream of the firefly luciferase cDNA and the recombinant constructs were transformed into cultured neuroblast cells and HEK-293 *** promoter activities were assessed by luciferase *** untranslated exon（228 bp, 197 bp and 50 bp respectively） were dispersed in 5’flanking region of human SCN3A and the TSS was located at 27.5 kb upstream of the first coding exon.A potential Initiator（INR）,an important element for general eukaryotic core promoter, was found at TSS.A mimic-TATA and a downstream regulatory element（DRE） were found at the -25 bp site and the +23 bp site,*** experiments demonstrated that the 1.1-kb genomic fragment was active in neuronal cell（SHSY5Y） and was inactive in non-

关键词： promoter transcription regulatory element sodium channel SCN3A

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Molecular Plant 2016年第3期9卷 428-436页

作者： J. Bohm S. Scherzer S. Shabala E. Krol E. Neher T.D. Mueller R. Hedrich Julius-yon-Sachs Institute Department for Molecular Plant Physiology and Biophysics University of Wurzburg Julius-von-Sachs Platz 2 97082 Wurzburg Germany School of Land and Food University of Tasmania Hobart TAS 7001 Australia Zoology Department College of Science King Saud University PO Box 2455 Riyadh 11451 Saudi Arabia Department for Membrane Biophysics Max Planck Institute for Biophysical Chemistry 37077 Goettingen Germany

The animal diet of the carnivorous Venus flytrap, Dionaea muscipula, contains a sodium load that enters the capture organ via an HKT1-type sodium channel, expressed in special epithelia cells on the inner trap lobe surface. DmHKT1 expression and sodium uptake activity is induced upon prey contact. Here, we analyzed the HKT1 properties required for prey sodium osmolyte management of carnivorous Dionaea. Analyses were based on homology modeling, generation of model-derived point mutants, and their functional testing in Xenopus oocytes. We showed that the wild-type HKT1 and its Na^＋- and K^＋-permeable mutants function as ion channels rather than K^＋ transporters driven by proton or sodium gradients. These structural and biophysical features of a high-capacity, Na^＋-selective ion channel enable Dionaea glands to manage prey-derived sodium loads without confounding the action potential-based information management of the flytrap.

关键词： sodium channel HKT1 Dionaea muscipula action potential glands sodium uptake