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whole-cell recordings of voltage-gated Calcium,Potassium and Sodium currents in acutely isolated hippocampal pyramidal neurons

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Journal of Nanjing Medical University 2009年第2期23卷 122-126页

作者： Shuyun Huang Qing Cai Weitian Liu Xiaoling Wang Tao Wang Department of physiology Tianjin University of Traditional Medicine Tianjin 300193 China Department of cerebral surgery People Hospital of FengRun Tangshan 064000 China

Objective：To record Calcium, Potassium and Sodium currents in acutely isolated hippocampal pyramidal neurons. Methods：Hippocampal CA3 neurons were freshly isolated by 1 mg protease/3 ml SES and mechanical trituration with polished pipettes of progressively smaller tip diameters. Patch clamp technique in whole-cell mode was employed to record voltage-gated channel currents. Results：The procedure dissociated hippocampal neurons, preserving apical dendrites and several basal dendrites, without impairing the electrical characteristics of the neurons. whole-cell patch clamp configuration was successfully used to record voltage-gated Ca^2＋ currents, delayed rectifier K^＋ current and voltage-gated Na^＋ currents. Conclusion：Protease combined with mechanical trituration may be used for the dissociation of neurons from rat hippocampus. Voltage-gated channels currents could be recorded using a patch clamp technique.

关键词： patch clamp hippocampus voltage-gated channels whole-cell

Significantly improved oxidation of bio-based furans into furan carboxylic acids using substrate-adapted whole cells

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Journal of Energy Chemistry 2020年第2期29卷 20-26页

作者： Mao Wen Xue-Ying Zhang Min-Hua Zong Ning Li School of Food Science and Engineering South China University of TechnologyGuangzhou 510640GuangdongChina

Furan carboxylic acids are important building blocks in polymer and fine chemical industries. In this work, a simple substrate adaptation strategy was applied to improve the catalytic performances of Comamonas testosteroni SC1588 cells for the synthesis of various furan carboxylic acids. It was found that biocatalytic synthesis of 5-hydroxymethyl-2-furancarboxylic acid(HMFCA) was substantially promoted by adding histidine and increasing cell concentrations. HMFCA was produced in a quantitative yield from200 m M HMF in 24 h. Besides, the HMFCA yields of 71%–81% were achieved with the substrate concentrations up to 250–300 m M. It was firstly found that 4-tert-butylcatechol(TBC), as the stabilizer present in HMF, exerted a significantly detrimental effect on whole-cell catalytic synthesis of HMFCA at high substrate concentrations(more than 130 m M). In addition, a variety of furan carboxylic acids such as2-furoic acid, 5-methyl-2-furancarboxylic acid and 5-methoxymethyl-2-furancarboxylic acid were synthesized with the yields up to 98%.

关键词： 5-hydroxymethylfurfural 5-hydroxymethyl-2-furancarboxylic acid Oxidation Substrate adaptation strategy whole-cell biocatalysis

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 *** investigated whether the increase of NKCC1 and KCC2 is associated with peripheral pain transmission in dorsal root ganglion *** this aim,rats with persistent hyperalgesia were randomly divided into four *** in the control group received no treatment,and the rat sciatic nerve was only exposed in the sham *** 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 *** the experiment measuring thermal withdrawal latency,bumetanide (15 mg/kg) was intravenously *** 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 *** Hargreaves test was conducted to detect changes in thermal hyperalgesia in *** found that the thermal withdrawal latency of rats was significantly decreased on days 7,14,and 21 after model *** intravenous injection of bumetanide,the reduction in thermal retraction latency caused by model establishment was significantly *** 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 *** immune response or protein expression of KCC2 was observed in dorsal root ganglion neurons before and after model *** 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 *** found th

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

Enantioselective Resolution of (±)-l-Phenylethyl Acetate by Using the whole cells of Deep-sea Bacterium Bacillus sp. DL-2

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Chemical Research in Chinese Universities 2019年第5期35卷 792-798页

作者： DONG Lu XU Yongkai ZHANG Yun SUN Aijun HU Yunfeng CAS Key Laboratory of Tropical Marine Bio-resources and Ecology South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhou 510301P.R.China University of Chinese Academy of Sciences Beijing 100049P.R.China Guangdong Key Laboratory of Marine Materia Medica South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhou 510301P.R.China Affiliated Hospital of Shandong University of Traditional Chinese Medicine Jinan 250014P.R.China

Bacillus sp. DL-2 was isolated from the deep sea of the Western Pacific and further utilized as novel biocatalysts to efficiently asymmetrically hydrolyze (±)-1-phenylethyl acetate. After the optimization of hydrolytic reactions, chiral chemicals (R)1-phenylethanol and (S)-l-phenylethyl acetate were obtained with high optical purities (96% and 99.8%, respectively). Our research is about the asymmeric hydrolysis of (±)-1-phenylethyl acetate using whole-cell biocatalysts. In addition, the optical purity of (S)-l-phenylethyl acetate generated through the kinetic resolution of (±)-1-phenylethyl acetate using the whole-cells of Bacillus sp. DL-2 was the highest report so far. Using the whole cells of deep sea bacterium Bacillus sp. DL-2 as the biocatalysts is an enviromnentally friendly method and will play critical roles in industrial asymmetric synthesis.

关键词： Deep-sea microorganism whole-cell biocatalyst Asymmetric hydrolysis (R)-1-Phenylethanol (S)-1-Phenylethyl acetate

Effects of Lizhong Tang on cultured mouse small intestine interstitial cells of Cajal

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World Journal of Gastroenterology 2013年第14期19卷 2249-2255页

作者： Min Woo Hwang Jung Nam Kim Ho Jun Song Bora Lim Young Kyu Kwon Byung Joo Kim Department of Sasang Constitutional Medicine College of Korean MedicineKyung-hee University School of Korean Medicine Pusan National University

AIM:To investigate the effects of Lizhong Tang,an herbal product used in traditional Chinese medicine,on mouse small intestine interstitial cells of Cajal(ICCs).METHODS:Enzymatic digestions were used to dissociate ICCs from mouse small intestine *** ICCs were morphologically distinct from other cell types in culture and were identified using phase contrast microscopy after verification with anti c-kit antibody.A whole-cell patch-clamp configuration was used to record potentials(current clamp) from cultured *** of the experiments were performed at 30-32 ℃.RESULTS:ICCs generated pacemaker potentials,and Lizhong Tang produced membrane depolarization in current-clamp *** application of flufenamic acid(a nonselective cation channel blocker) abolished the generation of pacemaker potentials by Lizhong *** with thapsigargin(a Ca 2+-ATPase inhibi-tor in the endoplasmic reticulum) also abolished the generation of pacemaker potentials by Lizhong ***,pacemaker potentials were completely abolished in the presence of an external Ca 2+-free solution,and under this condition,Lizhong Tang induced membrane ***,When GDPβ-S(1 mmol/L) was in the pipette solution,Lizhong Tang still induced membrane *** addition,membrane depolarizations were not inhibited by chelerythrine or calphostin C,which are protein kinase C inhibitors,but were inhibited by U-73122,an active phospholipase C ***:These results suggest that Lizhong Tang might affect gastrointestinal motility by modulating pacemaker activity in interstitial cells of Cajal.

关键词： Interstitial cells of Cajal Lizhong Tang Motility Gastrointestinal tract whole-cell patch clamp configuration

Effects of La^(3+) on inward K^+ channels at plasma membr in guard cells

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Science China Chemistry 2005年第2期48卷 143-147页

作者： XUE Shaowu YANG Pin Institute of Molecular Science Chemical Biology and Molecular Engineering Laboratory of Ministry of Education Shanxi University Taiyuan 030006 China Institute of Molecular Science Chemical Biology and Molecular Engineering Laboratory of Ministry of Education Shanxi University Taiyuan 030006 China

The effects of La3+ on inward K+ channels at plasma membrane in vicia guard cells are investigated using the whole-cell patch-clamp recording mode. It is shown that La3+ on both sides of plasma membrane blocks inward K+ currents in a concentration- dependent manner, indicating that La3+ binding sites may exist on both sides of plasma membrane in guard cells in vicia. The dose response is fitted by the Michaelis-Menten relation characterized by an inhibitor constant Ki of 2.56±0.25 μmol·L?1 (outside membrane) and (1.18±0.11)×10?15 mol·L?1 (inside membrane). Intracellular La3+ has much stronger inhibitory effect on inward K+ currents than ex- tracellular La3+ does, suggesting there may exist stronger binding sites inside membrane than outside membrane. Since ion channel activities of guard cells directly affect plant stomatal movement and water status, our results imply that rare earth elements might have potential practical values in regulating plant water status and strengthening plant drought endurance.

关键词： whole-cell patch-clamp, guard cell, stomatal movements, inward K+ channels, La3+.

Study of transmembrane La^(3+) movement in rat ventricular myocytes by the patch-clamp technique

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Chinese Science Bulletin 2002年第18期47卷 1518-1522页

作者： YANG Pin, DU Huizhi & XUE ShaowuInstitute of Molecular Science, Shanxi University, Taiyuan 030006, China Institute of Molecular Science Shanxi University Taiyuan China

We have studied transmembrane La3+ movement in rat ventricular myocytes for the first time by using the whole-cell patch-clamp recording mode. La3+ (0.01-5.0 mmol/L) could not bring out inward currents through the L-type calcium channel in rat ventricular myocytes, while it could enter the cells by the same way carried by 1μmo1/L ionomycin. When the outward Na+ concentration gradient is formed, La3+ can enter the cells via Na-Ca exchange, and the exchange currents increase with the increase of external La3+ concentrations. But compared with Na-Ca exchange currents in the same concentration, the former is only 14%-38% of the latter. The patch-clamp experiment indicates that La3+ normally can not enter ventricular myocytes through L-type calcium channel, but it can enter the cells via Na-Ca exchange.

关键词： whole-cell patch-clamp recording ventricular myocyte L-type calcium channel Na-Ca exchange La3+ Ca2+.

Toosendanin-induced Ca^(2+) Conductance Increase in Neuroblastoma×Giioma Hybrid cells

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Chinese Science Bulletin 1993年第17期38卷 1489-1493页

作者：施玉樑 Furuya Kishio 王文萍 Terakawa Susumu 徐科 Yamagishi Shunichi Shanghai Institute of Physiology Academia Sinica National Institute for Physiological Sciences

Toosendanin (TSN), a triterpenoid derivative extracted from the bark of Melia toosendan Seib et Zucc and used in Chinese traditional medicine as an anthelmintic against ascaris, is a presynaptic neuromuscular blocker. Without affecting the nerve conduction, the muscle resting potential as well as acetylcholine (ACh) sensitivity of the muscle membrane at the end-plate region, TSN blocks the neuromuscular transmission selectively by inhibiting ACh release from the motor nerve terminals. It was interesting to notice

关键词： toosendanin neuroblastoma Ca2+ conductance membrane potential whole-cell recording.