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Identification of SNARE proteins in fish—Tilapia Oreochromis niloticus

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Acta Oceanologica Sinica 2007年第1期26卷 159-163,164页

作者： HUANG Xiaohang LAM Patrick P L LIN Xuezheng LIU Chenlin BIAN Ji gaisano herbert Key Laboratory of Marine Bioactive Substances First Institute of Oceanography State Oceanic Administration Qingdao 266061 China Department of Physiology and Medicine University of Toronto Toronto M5S 1A8 Canada

SNARE proteins are a group of membrane-associated proteins involved in exocytosis, secretion and membrane trafficking events in eukaryotic cells. Research on SNARE protein biology has become a more attractive field in recent years, which is applied to marine biology specifically to the fish Tilapia （Oreochromis niloticus）. Plasma membrane fractions of different tissues of Tilapia, including brain, liver-pancreas, intestine, skin and muscle, were extracted, and immuno-decorated with isoform-specific antibodies to the SNARE families and associated proteins. The presence of Syntaxins - 1 A, 2 and 3, SNAP - 23 and SNAP - 25, VAMP - 2, Munc - 18 - 1 and Munc - 13 in the brain was identified, which were differentially distributed in the other organ tissues of the fish Tilapia. The distinct distribution of SNARE and associated proteins will serve as the basis for further investigation into their special secretory function in these tissues of the fish.

关键词： SNARE proteins exocytosis secretion fish

花鲈(Lateolabrax japonicus)VAMP-2基因的特征研究

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中国科学（C辑） 2006年第4期36卷 334-339页

作者：陈魁黄晓航包振民 gaisano herbert 中国海洋大学海洋生命学院国家海洋局第一海洋研究所青岛266061 Department of Medicine and Physiology University of TorontoTorontoM5S1A8Canada

应用反转录PCR和锚定PCR方法克隆了全长715个碱基的花鲈(Lateolabrax japonicus)SPV2基因,证明其编码110个氨基酸的VAMP-2蛋白.运用生物信息学研究工具进行的一级结构分析显示:SPV2蛋白含有SNARE基序及其核心精氨酸残基,其序列N端有一N... 详细信息

应用反转录PCR和锚定PCR方法克隆了全长715个碱基的花鲈(Lateolabrax japonicus)SPV2基因,证明其编码110个氨基酸的VAMP-2蛋白.运用生物信息学研究工具进行的一级结构分析显示:SPV2蛋白含有SNARE基序及其核心精氨酸残基,其序列N端有一N-糖肽键的糖基化位点,C端有一跨膜结构域.高级结构预测揭示SPV2蛋白的主要空间构象是一个组成SNARE基序的α螺旋卷曲骨架,其主要细节都与X晶体衍射方法得到的大鼠VAMP-2结构相一致.半定量PCR结果表明:在mRNA水平上,VAMP-2基因在鱼体各组织中广泛表达,其中以脑组织最为突出,而肌肉组织则相对最少.

关键词：花鲈 VAMP-2蛋白 SNARE蛋白生物信息学分析

Nitric oxide activation of a potassium channel (BK_(Ca)) in feline lower esophageal sphincter

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World Journal of Gastroenterology 2010年第46期16卷 5852-5860页

作者： Marie-Claude L'Heureux Ahmad Muinuddin herbert Y gaisano Nicholas E Diamant Department of Physiology University of Toronto Ahmad Muinuddin Nicholas E DiamantToronto Western Research InstituteUniversity Health Network Department of Medicine University of Toronto

AIM:To assess the effect of nitric oxide (NO) on the large conductance potassium channel (BKCa) in isolated circular (CM) and sling (SM) muscle cells and muscle strips from the cat lower esophageal sphincter (LES) to determine its regulation of resting tone and ***:Freshly enzymatically-digested and isolated circular smooth muscle cells were prepared from each LES *** study outward K + currents,the perforated patch clamp technique was *** assess LES resting tone and relaxation,muscle strips were mounted in perfused organ ***:(1) Electrophysiological recordings from isolated cells:(a) CM was more depolarized than SM (-39.7 ± 0.8mV vs-48.1 ± 1.6 mV,P 0.05);(b) The NO donor sodium nitroprusside (SNP) increased outward currents only in CM (25.9 ± 1.9 to 46.7 ± 4.2 pA/pF,P 0.05);(c) SNP added in the presence of the BK Ca antagonist iberiotoxin (IbTX) produced no increase in the outward current in CM (17.0 ± 2.8 vs 13.7 ± 2.2,P > 0.05);and (d) L-NNA caused a small insignificant inhibition of outward K + currents in both muscles;and (2) Muscle strip studies:(a) Blockade of the nerves with tetrodotoxin (TTX),or BK Ca with IbTX had no significant effect on resting tone of either muscle;and (b) SNP reduced tone in both muscles,and was unaffected by the presence of TTX or ***:Exogenous NO activates BK Ca only in CM of the ***,as opposed to other species,exogenous NO-induced relaxation is predominantly by a non-BK Ca mechanism,and endogenous NO has minimal effect on resting tone.

关键词： Circular smooth muscle Feline K+ channel Lower esophageal sphincter Nitric oxide Sling Tone

Characterization of SNAP-25 gene from marine teleostean, Lateolabrax japonicus

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Chinese Journal of Oceanology and Limnology 2007年第4期25卷 378-385页

作者：陈魁黄晓航柴迎梅 herbert Y. gaisano Institute of Oceanology Chinese Academy of Sciences The First Institute of Oceanography State Oceanic Administration Department of Physiology University of Toronto Toronto M5S 1A8 Canada

The t-SNARE protein SNAP-25 (synaptosome-associated protein of 25 kDa) plays an essential role in regulating fusion between the vesicle and plasma membranes during exocytosis. To clone and characterize SNAP-25 gene, the first step in the functional study of SNARE proteins in marine teleostean, was to obtain the cDNA of sea perch SNAP-25 (SPsn25) by RT-PCR and RACE-PCR amplification of a Japanese sea perch. The full-length cDNA of 831bp contains a CDS of 615 bp, coding 204 amino acid residues, and a 5′UTR of 219bp. Bioinformatic analysis revealed that SPsn25 corresponds with SNAP-25a isoform and shares 91.1% identity with SNAP-25a of a goldfish and a zebrafish. The SPsn25 expression in both mRNA and protein levels in the Japanese sea perch had been identified through semi-quantitative RT-PCR and Western Blot assay. Together, these data again confirmed the nerve tissue specificity of the fish SNAP-25 gene expression.

关键词： Lateolabrax japonicus SNARE SNAP-25

Munc18/SNARE proteins’ regulation of exocytosis in guinea pig duodenal Brunner’s gland acini

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World Journal of Gastroenterology 2008年第15期14卷 2314-2322页

作者： Laura I Cosen-Binker Gerry P Morris Stephen Vanner herbert Y gaisano Department of Medicine University of Toronto Toronto Ontario M5S 1A8 Canada Gastrointestinal Diseases Research Unit Queen's University Kingston Ontario KL7 5G2 Canada

AIM: To examine the molecular mechanism of exocytosis in the Brunner’s gland acinar cell. METHODS: We used a submucosal preparation of guinea pig duodenal Brunner’s gland acini to visualize the dilation of the ductal lumen in response to cholinergic stimulus. We correlated this to electron microscopy to determine the extent of exocytosis of the mucin-filled vesicles. We then examined the behavior of SNARE and interacting Munc18 proteins by confocal microscopy. RESULTS: One and 6 μmol/L carbachol evoked a dose- dependent dilation of Brunner’s gland acini lumen, which correlated to the massive exocytosis of mucin. Munc18c and its cognate SNARE proteins Syntaxin-4 and SNAP-23 were localized to the apical plasma membrane, and upon cholinergic stimulation, Munc18c was displaced into the cytosol leaving Syntaxin-4 and SNAP-23 intact. CONCLUSION: Physiologic cholinergic stimulation induces Munc18c displacement from the Brunner’s gland acinar apical plasma membrane, which enables apical membrane Syntaxin-4 and SNAP-23 to form a SNARE complex with mucin-filled vesicle SNARE proteins to affect exocytosis.

关键词： Apical exocytosis Brunner's gland acini Munc18c Syntaxin-4 Carbachol

Chronic stress sensitizes rats to pancreatitis induced by cerulein:Role of TNF-α

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World Journal of Gastroenterology 2010年第44期16卷 5565-5581页

作者： Marcelo G Binker Andres A Binker-Cosen Daniel Richards herbert Y gaisano Rodica H de Cosen Laura I Cosen-Binker CBRHC Research Health Center Department of Medicine and Physiology University of Toronto

AIM:To investigate chronic stress as a susceptibility factor for developing pancreatitis,as well as tumor necrosis factor-α (TNF-α) as a putative ***:Rat pancreatic acini were used to analyze the influence of TNF-α on submaximal (50 pmol/L) cholecystokinin (CCK) *** restraint (4 h every day for 21 d) was used to evaluate the effects of submaximal (0.2 μg/kg per hour) cerulein stimulation on chronically stressed ***:In vitro exposure of pancreatic acini toTNF-α disorganized the actin *** was further increased by TNF-α/CCK treatment,which additionally reduced amylase secretion,and increased trypsin and nuclear factor-κB activities in a protein-kinase-C δ and ε-dependent ***-α/CCK also enhanced caspases' activity and lactate dehydrogenase release,induced ATP loss,and augmented the ADP/ATP *** vivo,rats under chronic restraint exhibited elevated serum and pancreatic TNF-α ***,pancreatic,and lung inflammatory parameters,as well as caspases' activity in pancreatic and lung tissue,were substantially enhanced in stressed/cerulein-treated rats,which also experienced tissues' ATP loss and greater ADP/ATP *** examination revealed that stressed/cerulein-treated animals developed abundant pancreatic and lung edema,hemorrhage and leukocyte infiltrate,and pancreatic *** severity was greatly decreased by treating animals with an anti-TNF-αantibody,which diminished all inflammatory parameters,histopathological scores,and apoptotic/necrotic markers in stressed/cerulein-treated ***:In rats,chronic stress increases susceptibility for developing pancreatitis,which involves TNF-α sensitization of pancreatic acinar cells to undergo injury by physiological cerulein stimulation.

关键词： Pancreatitis Stress Tumor necrosis factor-α

VAMP8在beta细胞分泌和增殖中的调节作用

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VAMP8在beta细胞分泌和增殖中的调节作用