Neuroprotocols最新文献_第9页

Use of U-73122 as an Inhibitor of Phospholipase C-Dependent Processes U-73122作为磷脂酶C依赖过程抑制剂的应用

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/ncmn.1993.1046

Bleasdale John E., Fisher Stephen K.

{"title":"Use of U-73122 as an Inhibitor of Phospholipase C-Dependent Processes","authors":"Bleasdale John E., Fisher Stephen K.","doi":"10.1006/ncmn.1993.1046","DOIUrl":"https://doi.org/10.1006/ncmn.1993.1046","url":null,"abstract":"<div>1-[6-[[17β-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]-hexyl]1H-pyrrole-2,5-dione (U-73122) is an aminosteroid that was identified initially as a potent inhibitor of platelet activation by receptor-specific agonists. U-73122 inhibits receptor-coupled generation of inositol 1,4,5-trisphosphate (but not cyclic AMP) and intracellular mobilization of Ca 2+ in a variety of cell types. U-73122 inhibits phosphoinositide-specific phospholipase C (PI-PLC) activity in cell-free systems, but exhibits little or no direct inhibition of phospholipases A2 and D. Structure-activity analysis revealed that the maleimide group of U-73122 is essential, but not sufficient, for inhibitory activity. The succinimide analog of U-73122 (U-73343) has negligible inhibitory activity and is a useful control compound. On the basis of information derived from the use of U-73122 in a variety of cell types, procedures for storing, dissolving, and presenting U-73122 to cells are recommended. While knowledge of the mechanism of action of U-73122 would extend the utility of this compound, U-73122 has already been employed successfully to examine PI-PLC involvement in a variety of cellular processes. The application of U-73122 in an investigation of muscarinic receptor sequestration in SK-N-SH neuroblastoma cells is illustrated.</div>","PeriodicalId":100951,"journal":{"name":"Neuroprotocols","volume":"3 2","pages":"Pages 125-133"},"PeriodicalIF":0.0,"publicationDate":"1993-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/ncmn.1993.1046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72117730","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 53

Cytidine-Diphosphate Diacylglycerol Labeling as an Index of Inositol Lipid-Mediated Signal Transduction in Brain and Neural Cells 胞苷-二磷酸二酰基甘油标记作为肌醇脂质介导的脑和神经细胞信号转导的指标

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/NCMN.1993.1043

A. Heacock, E. Stubbs, B. Agranoff

{"title":"Cytidine-Diphosphate Diacylglycerol Labeling as an Index of Inositol Lipid-Mediated Signal Transduction in Brain and Neural Cells","authors":"A. Heacock, E. Stubbs, B. Agranoff","doi":"10.1006/NCMN.1993.1043","DOIUrl":"https://doi.org/10.1006/NCMN.1993.1043","url":null,"abstract":"Abstract A method for assessing stimulated phosphoinositide turnover by measurement of the liponucleotide CDP-diacylglycerol is presented. The phosphoinositide signal transduction pathway consists of a sequence of reactions in which the second messengers Inositol 1,4,5-triphosphate and diacylglycerol are recycled back to phosphatidylinositol (PtdIns), which then serves to replenish the initial hydrolyzed substrate, phosphatidylinositol 4,5-bis-phosphate. Receptor-stimulated inositol lipid turnover is most commonly assessed by measurement of the accumulation of [ 3 H]inositol-labeled inositol phosphates in the presence of Li + . The latter blocks Inositol monophosphatase and thus can lead to a depletion of intracellular inositol. Because inositol is required for resynthesis of PtdIns, the immediate precursor of PtdIns, CDP-diacylglycerol, also accumulates in the presence of agonist and Li + . Measurement of radiolabeling of this liponucleotide following Incorporation of [ 3 H]cytidine thus forms the basis for an alternative assay for Inositol lipid turnover. The general applicability of this method may be limited, since, In brain slices, not all receptors exhibit CDP-diacylglycerol responses that are consistent with their inositol phosphate responses. In addition, in cultured neural cells, growth in inositol-free, chemically defined medium is required to maximize the Li + -dependent CDP-diacylglycerol response. A major advantage of this method may be its ability to provide insight Into the regulation of phosphoinositide turnover since this method uniquely reflects slowing of the regenerative cycle. Such in vitro studies may have relevance to the in vivo action of Li + as a psychotherapeutic agent.","PeriodicalId":100951,"journal":{"name":"Neuroprotocols","volume":"74 1","pages":"103-106"},"PeriodicalIF":0.0,"publicationDate":"1993-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79607041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Use of an Inhibitor of Glucosylceramide Synthesis, D-1-Phenyl-2-decanoylamino-3-morpholino-1 -propanol 葡萄糖神经酰胺合成抑制剂d -1-苯基-2-癸烷氨基-3-氨基-1-丙醇的应用

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/NCMN.1993.1048

N. Radin, J. Shayman

{"title":"Use of an Inhibitor of Glucosylceramide Synthesis, D-1-Phenyl-2-decanoylamino-3-morpholino-1 -propanol","authors":"N. Radin, J. Shayman","doi":"10.1006/NCMN.1993.1048","DOIUrl":"https://doi.org/10.1006/NCMN.1993.1048","url":null,"abstract":"Abstract Methods for the synthesis, purification, and use of an inhibitor of glucosylceramide synthesis (PDMP or D-1-phenyl-2-decanoylamino-3-morpholino-1-propanol) are given. The inhibitor is effective with a variety of cells and animals in producing a depletion of glucosylceramide. Because this cerebroside is the precursor of hundreds of other glycolipids, depletion of all of these compounds also takes place as each one is degraded by hydrolases. Use of PDMP causes accumulation of ceramide, the lipoidal precursor of glucosylceramide. Some of this simple sphingolipid is diverted to the synthesis of sphingomyelin and some, via hydrolysis, to formation of sphingols (sphingoid bases). The above changes in the biosynthesis of glycolipids result in pronounced effects on cells: slowing of growth, increased activity of the phospholipase C that catalyzes phosphatidylinositol bisphosphate hydrolysis, accumulation of N,N -dimethylsphingosine (an inhibitor of protein kinase C), accumulation of diacyiglycerol (an activator of PKC), and reduction of the ability to bind to extracellular matrix proteins. PDMP is rapidly absorbed and released by cells. In mice, it is metabolized by a microsomal monooxygenase and the products are excreted. The degradative process can be blocked by inhibitors of cytochrome P-450, such as piperonyl butoxide, cimetidine, and fluconazole. Understanding these properties permits the use of PDMP in both in vitro and in vivo studies in which glycolipids may exhibit important biological effects. Two recent examples of the in vitro and in vivo use of PDMP are provided.","PeriodicalId":100951,"journal":{"name":"Neuroprotocols","volume":"51 1","pages":"145-155"},"PeriodicalIF":0.0,"publicationDate":"1993-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78903333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

α-Thrombin-Stimulated 1,2-Diacylglycerol Formation: The Relationship between Phospholipid Hydrolysis and Protein Kinase C Activation α-凝血酶刺激1,2-二酰甘油形成：磷脂水解与蛋白激酶C激活的关系

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/ncmn.1993.1042

Leach Karen L., Raben Daniel M.

引用次数: 1

Assessment of Neuronal Phosphoinositide Turnover and Its Disruption by Lithium 锂离子对神经元磷酸肌苷转换及其破坏的影响

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/NCMN.1993.1047

R. Challiss, S. Jenkinson, R. Mistry, I. Batty, S. Nahorski

引用次数: 17

Optimizing Biological Activity of Lipophilic Compounds in Cultured Cells by Improving Delivery: Theoretical and Practical Considerations 通过改善递送来优化培养细胞中亲脂化合物的生物活性:理论和实践考虑

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/NCMN.1993.1050

S. Buxser

{"title":"Optimizing Biological Activity of Lipophilic Compounds in Cultured Cells by Improving Delivery: Theoretical and Practical Considerations","authors":"S. Buxser","doi":"10.1006/NCMN.1993.1050","DOIUrl":"https://doi.org/10.1006/NCMN.1993.1050","url":null,"abstract":"Abstract The use of lipophilic compounds in the study of signal transduction and other cellular processes is often complicated by the need to deliver compounds that are minimally soluble in water to cells in culture. A typical approach is to dissolve the lipophilic compound at high concentration in organic solvent and then to dilute the solvent in aqueous medium. However, such an approach usually results in precipitation of the compound in the aqueous medium and may minimize delivery of the compound to cells. Three techniques to keep the lipophilic compound In a nonprecipitated and stable form that will be readily available to the cells are described. They involve the use of protein carriers, liposomes, and emulsions. Particular attention is given to the use of emulsions, since this technique combines the advantages of a high capacity for lipophilic compounds, ease of assembly, and minimum contact between the lipophilic compound and water. Although the techniques differ with respect to the mechanics of combining the compound and the carrier, they all consist of a two-phase system dependent on partitioning between the carrier and the cells. Due to the need to take partitioning into account, all of these techniques differ from homogeneous solution-phase delivery. Therefore, in addition to descriptions of the techniques, criteria for both experimental design and analysis of data generated using two-phase systems are presented. In combination, the use of methods appropriate for delivery of lipophilic compounds to cells and the application of relevant calculations and analytical procedures provide the means necessary for design and Interpretation of experiments using lipophilic compounds.","PeriodicalId":100951,"journal":{"name":"Neuroprotocols","volume":"26 1","pages":"165-174"},"PeriodicalIF":0.0,"publicationDate":"1993-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78237604","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Cytidine-Diphosphate Diacylglycerol Labeling as an Index of Inositol Lipid-Mediated Signal Transduction in Brain and Neural Cells 二磷酸胞苷二酰甘油标记作为脑和神经细胞肌醇脂质介导的信号转导指标

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/ncmn.1993.1043

Heacock Anne M., Stubbs Jr., Evan B., Agranoff Bernard W.

{"title":"Cytidine-Diphosphate Diacylglycerol Labeling as an Index of Inositol Lipid-Mediated Signal Transduction in Brain and Neural Cells","authors":"Heacock Anne M., Stubbs Jr., Evan B., Agranoff Bernard W.","doi":"10.1006/ncmn.1993.1043","DOIUrl":"https://doi.org/10.1006/ncmn.1993.1043","url":null,"abstract":"<div>A method for assessing stimulated phosphoinositide turnover by measurement of the liponucleotide CDP-diacylglycerol is presented. The phosphoinositide signal transduction pathway consists of a sequence of reactions in which the second messengers Inositol 1,4,5-triphosphate and diacylglycerol are recycled back to phosphatidylinositol (PtdIns), which then serves to replenish the initial hydrolyzed substrate, phosphatidylinositol 4,5-bis-phosphate. Receptor-stimulated inositol lipid turnover is most commonly assessed by measurement of the accumulation of [3H]inositol-labeled inositol phosphates in the presence of Li+. The latter blocks Inositol monophosphatase and thus can lead to a depletion of intracellular inositol. Because inositol is required for resynthesis of PtdIns, the immediate precursor of PtdIns, CDP-diacylglycerol, also accumulates in the presence of agonist and Li+. Measurement of radiolabeling of this liponucleotide following Incorporation of [3H]cytidine thus forms the basis for an alternative assay for Inositol lipid turnover. The general applicability of this method may be limited, since, In brain slices, not all receptors exhibit CDP-diacylglycerol responses that are consistent with their inositol phosphate responses. In addition, in cultured neural cells, growth in inositol-free, chemically defined medium is required to maximize the Li+ -dependent CDP-diacylglycerol response. A major advantage of this method may be its ability to provide insight Into the regulation of phosphoinositide turnover since this method uniquely reflects slowing of the regenerative cycle. Such in vitro studies may have relevance to the in vivo action of Li+ as a psychotherapeutic agent.</div>","PeriodicalId":100951,"journal":{"name":"Neuroprotocols","volume":"3 2","pages":"Pages 103-106"},"PeriodicalIF":0.0,"publicationDate":"1993-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/ncmn.1993.1043","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72117727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Assay of Phospholipase D as a Neuronal Receptor-Effector Mechanism 磷脂酶D作为神经元受体效应机制的测定

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/ncmn.1993.1049

Boarder M.R., Purkiss J.R.

引用次数: 4

Use of an Inhibitor of Glucosylceramide Synthesis, D-1-Phenyl-2-decanoylamino-3-morpholino-1 -propanol 葡萄糖神经酰胺合成抑制剂D-1-苯基-2-癸酰氨基-3-吗啉-1-丙醇的应用

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/ncmn.1993.1048

Radin Norman S., Shayman James A.

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引用次数: 4

Assay of Phospholipase D as a Neuronal Receptor-Effector Mechanism 磷脂酶D作为神经元受体-效应机制的研究

Neuroprotocols Pub Date : 1993-10-01 DOI: 10.1006/NCMN.1993.1049

M. Boarder, J. Purkiss

引用次数: 4