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Regulatory effects of anandamide on intracellular Ca^(2+) concentration increase in trigeminal ganglion neurons

Neural Regeneration Research 2014年第8期9卷 878-887页

作者： Yi Zhang Hong Xie Gang Lei Fen Li Jianping Pan Changjin Liu Zhiguo Liu Lieju Liu Xuehong Cao Department of Anesthesiology Tongji Hospital Affiliated to Tongji Medical CollegeHuazhong University of Science and Technology Jingzhou Central Hospital Department of Physiology Tongji Medical CollegeHuazhong University of Science and Technology Department of Bioengineering Wuhan Institute of Engineering

Activation of cannabinoid receptor type 1 on presynaptic neurons is postulated to suppress neu- ~ ~ ~ 2＋ ~ ~ 2＋ rotransmlsslon by decreasing Ca reflux through high voltage-gated Ca channels. However, recent studies suggest that cannabinoids which activate cannabinoid receptor type 1 can increase neurotransmitter release by enhancing Ca2＋ influx in vitro. The aim of the present study was to investigate the modulation of intracellular Ca2＋ concentration by the cannabinoid receptor type 1 agonist anandamide, and its underlying mechanisms. Using whole cell voltage-damp and calcium imaging in cultured trigeminal ganglion neurons, we found that anandamide directly caused Ca2＋ influx in a dose-dependent manner, which then triggered an increase of intracellular Ca2＋ concentration. The cyclic adenosine and guanosine monophosphate-dependent protein kinase systems, but not the protein kinase C system, were involved in the increased intracellular Ca2＋concentration by anandamide. This result showed that anandamide increased intracellu- lar Ca2＋ concentration and inhibited high voltage-gated Ca2＋ channels through different signal transduction pathways.

关键词： nerve regeneration trigeminal ganglion neurons endocannabinoids anandamide can-nabinoid receptor type 1 voltage-dependent calcium channels vanilloid receptor patch-damp tech-nique calcium cyclic adenosine monophosphate protein kinase protein kinase C NIH grant neuralregeneration

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Action of aluminum on high voltage-dependent calcium current and its modulation by ginkgolide B

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Acta Pharmacologica Sinica 2005年第5期26卷 539-545页

作者： Jurgen HESCHLER Cologne D-50931 Germany Neurophysiology University of Cologne

Aim:To investigate the effect of aluminum(Al)on high voltage-dependent cal-cium current(IHVA)and its modulation by ginkgolide B(Gin B).Methods:Thewhole-cell,patch-clamp technique was used to record IHVAfrom acutely isolatedhippocampal CA1 pyramydal neurons in ***:A1 0.1 mmol/L(lowconcentration)reduced IHVA;A1 0.75 and 1.0 mmol/L(high concentrations)in-creased IHVA,and Al decreased and increased IHVAat intermediate concentrationsof 0.25 and 0.5 mmol/*** increase of IHVAby Al 1.0 mmol/L was enhanced by theadenylyl cyclase(AC)agonist forskolin and was partly abolished by the cyclicadenosine monophosphate(cAMP)-dependent protein kinase A(PKA)antago-nist H-89,whereas the decrease observed with Al 0.1 mmol/L was neither reversedby forskolin nor affected by *** B had no effect on IHVAin normal neurons,but canceled the increase in IHVAby 1.0 mmol/L ***:The resultsindicate that the mechanism of Al affecting IHVAdiffers at different concentrations,and this may be attributed to its complex *** B could prevent neuronsfrom injury by inhibiting calcium influx.

关键词： hippocampus aluminum voltage-dependent calcium channels ginkgolide B patch-clamp technique

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Induction of pancreatic duct cells of neonatal rats into insulin-producing cells with fetal bovine serum: A natural protocol and its use for patch clamp experiments

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World Journal of Gastroenterology 2005年第44期11卷 6968-6974页

作者： San-Hua Leng Fu-Er Lu Institute of Integrative Traditional Chinese and Western Medicine Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China

AIM： To induce the pancreatic duct cells into endocrine cells with a new natural protocol for electrophysiological study. METHODS： The pancreatic duct cells of neonatal rats were isolated, cultured and induced into endocrine ceils with 15% fetal bovine serum for a period of 20 d. During this period, insulin secretion, MTT value, and morphological change of neonatal and adult pancreatic islet cells were comparatively investigated. Pancreatic β-cells were identified by morphological and electrophysiological characteristics, while ATP sensitive potassium channels （KATP）, voltage-dependent potassium channels （Kv）, and voltage-dependent calcium channels （KcA） in β-cells were identified by patch clamp technique. RESULTS： After incubation with fetal bovine serum, the neonatal duct cells budded out, changed from duct-like cells into islet clusters. In the first 4 d, MTT value and insulin secretion increased slowly （MTT value from 0.024 ±0.003 to　3.028±0.003, insulin secretion from 2.6±0.6 to 3.1±0.8 mIU/L）. Then MTT value and insulin secretion increased quickly from d 5 to d 10 （MTT value from 0.028 ±0.003 to 0.052±0.008, insulin secretion from 3.1±0.8 to 18.3±2.6 mIU/L）, then reached high plateau （MTT value 〉0.052±0.008, insulin secretion 〉18.3±2.6 mIU/L）. In contrast, for the isolated adult pancreatic islet cells, both insulin release and MTT value were stable in the first 4 d （MTT value from 0.029±0.01 to 0.031±0.011, insulin secretion from 13.9±3.1 to 14.3±3.3 mIU/L）, but afterwards they reduced gradually （MTT value 〈0.031 ±0.011, insulin secretion 〈8.2±1.5 mIU/L）, and the pancreatic islet cells became dispersed, broken or atrophied correspondingly. The differentiated neonatal cells were identified as pancreatic islet cells by dithizone staining method, and pancreatic β-cells were further identified by both morphological features and electrophysiological characteristics, i.e. the existence of recording currents from KATP, Kv, and KCA. CONCLUSION： Islet cells different

关键词： Pancreatic duct cells Pancreatic precursor cells Insulin-producing cells Patch clamp Experimental protocol ATP sensitive potassium channels voltagedependent potassium channels voltage-dependent calcium channels

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Pharmacological Investigation of voltage-dependent Ca^(2+) channels in Human Ejaculatory Sperm in vitro

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第5期26卷 607-609页

作者：李路刘继红李家贵叶章群 Department of Urology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

The types of the voltage-dependent calcium channels （VDCCs） in human ejaculatory sperm and the effects of calcium channel blocker （CCB） on human sperm motility parameters in vitro were investigated. The human sperm motility parameters in vitro in response to the pharmacological agents nifedipine （NIF, inhibitor of L-type VDCC） and ω-conotoxin （GVIA, inhibitor of N-type VDCC） were compared and analyzed statistically. The results showed that NIF （1, 5, 10 μmol/L） could not only significantly affect human sperm＇s shape but also spermatozoa motility after incubated at least 10 rain in vitro （P〈0.001）. GVIA （0.1, 0.5 and 1 μmol/L） could just only significantly affect human sperm＇s progressive motility （a %＋b %） after incubated for 20 min in vitro （P〈0.01）, but they both could not significantly affect spermic abnormality rate. It is suggested that L-type VDCC, non L-type VDCCs and isoform of L-type VDCC exist in the cell membrane of human sperm solely or together, and they participate in the spermic physiological processes especially the spermic motility.

关键词： voltage-dependent calcium channels calcium channel blocker human sperm motility in vitro

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