Trodusquemine is an aminosterol with a variety of biological and pharmacological functions, such as acting as an antimicrobial, stimulating body weight loss and interfering with the toxicity of proteins involved in the development of Alzheimer's and Parkinson's diseases. The mechanisms of interaction of aminosterols with cells are, however, still largely uncharacterized. Here, by using fluorescently labeled trodusquemine (TRO-A594 and TRO-ATTO565), we show that trodusquemine binds initially to the plasma membrane of living cells, that the binding affinity is dependent on cholesterol, and that trodusquemine is then internalized and mainly targeted to lysosomes after internalization. We also found that TRO-A594 is able to strongly and selectively bind to myelinated fibers in fixed mouse brain slices, and that it is a marker compatible with tissue clearing and light-sheet fluorescence microscopy or expansion microscopy. In conclusion, this work contributes to further characterize the biology of aminosterols and provides a new tool for nerve labeling suitable for the most advanced microscopy techniques.

Studying the trafficking of labeled trodusquemine and its application as nerve marker for light-sheet and expansion microscopy / Claudia Capitini, Luca Pesce, Giulia Fani, Giacomo Mazzamuto, Massimo Genovese, Alessandra Franceschini, Paolo Paoli, Giuseppe Pieraccini, Michael Zasloff, Fabrizio Chiti, Francesco S Pavone, Martino Calamai. - In: THE FASEB JOURNAL. - ISSN 0892-6638. - ELETTRONICO. - 38:(2022), pp. e22655-e22655. [10.1096/fj.202201276R]

Studying the trafficking of labeled trodusquemine and its application as nerve marker for light-sheet and expansion microscopy

Claudia Capitini;Luca Pesce;Giulia Fani;Giacomo Mazzamuto;Massimo Genovese;Alessandra Franceschini;Paolo Paoli;Giuseppe Pieraccini;Fabrizio Chiti;Francesco S Pavone;Martino Calamai
2022

Abstract

Trodusquemine is an aminosterol with a variety of biological and pharmacological functions, such as acting as an antimicrobial, stimulating body weight loss and interfering with the toxicity of proteins involved in the development of Alzheimer's and Parkinson's diseases. The mechanisms of interaction of aminosterols with cells are, however, still largely uncharacterized. Here, by using fluorescently labeled trodusquemine (TRO-A594 and TRO-ATTO565), we show that trodusquemine binds initially to the plasma membrane of living cells, that the binding affinity is dependent on cholesterol, and that trodusquemine is then internalized and mainly targeted to lysosomes after internalization. We also found that TRO-A594 is able to strongly and selectively bind to myelinated fibers in fixed mouse brain slices, and that it is a marker compatible with tissue clearing and light-sheet fluorescence microscopy or expansion microscopy. In conclusion, this work contributes to further characterize the biology of aminosterols and provides a new tool for nerve labeling suitable for the most advanced microscopy techniques.
2022
38
e22655
e22655
Claudia Capitini, Luca Pesce, Giulia Fani, Giacomo Mazzamuto, Massimo Genovese, Alessandra Franceschini, Paolo Paoli, Giuseppe Pieraccini, Michael Zasloff, Fabrizio Chiti, Francesco S Pavone, Martino Calamai
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1311884
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