The Lucigenin Assay: Measuring Anion Transport in Lipid Vesicles**

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Abstract

Synthetic anion transporters are developed to transport anions across lipid membranes with the long‐term perspective of biological applications. The lucigenin assay is a popular tool to study their transport of chloride and other anions in liposomes. It relies on the quenching of the fluorescence of encapsulated lucigenin by anions, which can be monitored by fluorescence spectroscopy. This article provides a tutorial introduction to the practical use and understanding of the lucigenin assay. It describes in detail how to use this assay to monitor chloride/nitrate antiport in liposomes, process and interpret the data, and solve common issues. Variations of the assay enabling the investigation of the transport of other anions and transport mechanisms are discussed. Furthermore, a zwitterionic analogue of lucigenin is demonstrated to have advantages for use in experiments over longer time scales, as it does not leak out of the liposomes, or when studying chloride uniport, as it avoids interference from antiport with nitrate that is present in commercial lucigenin.

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Superoxide Ion: Generation and Chemical Implications.

Maan Hayyan, Mohd Hashim, Inas AlNashef (2016)

Superoxide ion (O2(•-)) is of great significance as a radical species implicated in diverse chemical and biological systems. However, the chemistry knowledge of O2(•-) is rather scarce. In addition, numerous studies on O2(•-) were conducted within the latter half of the 20th century. Therefore, the current advancement in technology and instrumentation will certainly provide better insights into mechanisms and products of O2(•-) reactions and thus will result in new findings. This review emphasizes the state-of-the-art research on O2(•-) so as to enable researchers to venture into future research. It comprises the main characteristics of O2(•-) followed by generation methods. The reaction types of O2(•-) are reviewed, and its potential applications including the destruction of hazardous chemicals, synthesis of organic compounds, and many other applications are highlighted. The O2(•-) environmental chemistry is also discussed. The detection methods of O2(•-) are categorized and elaborated. Special attention is given to the feasibility of using ionic liquids as media for O2(•-), addressing the latest progress of generation and applications. The effect of electrodes on the O2(•-) electrochemical generation is reviewed. Finally, some remarks and future perspectives are concluded.

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Membrane lipidome of an epithelial cell line.

Hartmut Beug, Mathias J. Gerl, Kai Simons … (2011)

Tissue differentiation is an important process that involves major cellular membrane remodeling. We used Madin-Darby canine kidney cells as a model for epithelium formation and investigated the remodeling of the total cell membrane lipidome during the transition from a nonpolarized morphology to an epithelial morphology and vice versa. To achieve this, we developed a shotgun-based lipidomics workflow that enabled the absolute quantification of mammalian membrane lipidomes with minimal sample processing from low sample amounts. Epithelial morphogenesis was accompanied by a major shift from sphingomyelin to glycosphingolipid, together with an increase in plasmalogen, phosphatidylethanolamine, and cholesterol content, whereas the opposite changes took place during an epithelial-to-mesenchymal transition. Moreover, during polarization, the sphingolipids became longer, more saturated, and more hydroxylated as required to generate an apical membrane domain that serves as a protective barrier for the epithelial sheet.