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A pH-Sensitive Double Chromophore Fluorescent Dye for Live-Tracking of Lipophagy.
Engelhardt, Pascal M; Veronese, Matteo; Eryigit, Alpay A; Das, Anushka; Kaczmarek, Alexander T; Rugarli, Elena I; Schmalz, Hans-Günther.
Afiliação
  • Engelhardt PM; Department of Chemistry, University of Cologne, Greinstraße 4, 50939, Cologne, Germany.
  • Veronese M; Cluster of Excellence-Cellular Stress Responses in Aging-Associated Diseases (CECAD), Center of Molecular Medicine Cologne (CMMC), University of Cologne, Joseph-Stelzmann-Straße 26, 50931, Cologne, Germany.
  • Eryigit AA; Department of Chemistry, University of Cologne, Greinstraße 4, 50939, Cologne, Germany.
  • Das A; Cluster of Excellence-Cellular Stress Responses in Aging-Associated Diseases (CECAD), Center of Molecular Medicine Cologne (CMMC), University of Cologne, Joseph-Stelzmann-Straße 26, 50931, Cologne, Germany.
  • Kaczmarek AT; Cluster of Excellence-Cellular Stress Responses in Aging-Associated Diseases (CECAD), Center of Molecular Medicine Cologne (CMMC), University of Cologne, Joseph-Stelzmann-Straße 26, 50931, Cologne, Germany.
  • Rugarli EI; Cluster of Excellence-Cellular Stress Responses in Aging-Associated Diseases (CECAD), Center of Molecular Medicine Cologne (CMMC), University of Cologne, Joseph-Stelzmann-Straße 26, 50931, Cologne, Germany.
  • Schmalz HG; Department of Chemistry, University of Cologne, Greinstraße 4, 50939, Cologne, Germany.
Chemistry ; 30(30): e202400808, 2024 May 28.
Article em En | MEDLINE | ID: mdl-38506349
ABSTRACT
Lipid droplet (LD) degradation provides metabolic energy and important building blocks for various cellular processes. The two major LD degradation pathways include autophagy (lipophagy), which involves delivery of LDs to autolysosomes, and lipolysis, which is mediated by lipases. While abnormalities in LD degradation are associated with various pathological disorders, our understanding of lipophagy is still rudimentary. In this study, we describe the development of a lipophilic dye containing two fluorophores, one of which is pH-sensitive and the other pH-stable. We further demonstrate that this "Lipo-Fluddy" can be used to visualize and quantify lipophagy in living cells, in an easily applicable and protein label-free approach. After estimating the ability of compound candidates to penetrate LDs, we synthesized several BODIPY and (pH-switchable) rhodol dyes, whose fluorescence properties (incl. their photophysical compatibility) were analyzed. Of three Lipo-Fluddy dyes synthesized, one exhibited the desired properties and allowed observation of lipophagy by fluorescence microscopy. Also, this dye proved to be non-toxic and suitable for the examination of various cell lines. Moreover, a method was developed to quantify the lipophagy process using flow cytometry, which could be applied in the future in the identification of lipophagy-related genes or in the screening of potential drugs against lipophagy-related diseases.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Autofagia / Compostos de Boro / Gotículas Lipídicas / Corantes Fluorescentes Limite: Humans Idioma: En Revista: Chemistry Assunto da revista: QUIMICA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Alemanha

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Autofagia / Compostos de Boro / Gotículas Lipídicas / Corantes Fluorescentes Limite: Humans Idioma: En Revista: Chemistry Assunto da revista: QUIMICA Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Alemanha