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

Year: 2022

Authors: Capitini C., Pesce L., Fani G., Mazzamuto G., Genovese M., Franceschini A., Paoli P., Pieraccini G., Zasloff M., Chiti F., Pavone FS., Calamai M.

Autors Affiliation: Univ Florence, European Lab Nonlinear Spect LENS, Sesto Fiorentino, Italy; Univ Florence, Dept Phys, Sesto Fiorentino, Italy; Univ Florence, Dept Expt & Clin Biomed Sci, Sect Biochem, Florence, Italy; Natl Res Council CNR INO, Natl Inst Opt, Sesto Fiorentino, Italy; Univ Florence, CISM Mass Spectrometry Ctr, Sesto Fiorentino, Italy; Enterin Inc, Philadelphia, PA USA; Georgetown Univ, MedStar Georgetown Transplant Inst, Sch Med, Washington, DC USA.

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.

Journal/Review: FASEB JOURNAL

Volume: 36 (12)      Pages from: e22655-1  to: e22655-16

More Information: European Union & apos;s Horizon 2020 research and innovation programme, Grant/Award Number: 654148 Laserlab- Europe; General Hospital Corporation Center of the National Institutes of Health, Grant/Award Number: MH117023
KeyWords: intracellular trafficking; lipid membrane; nerve fiber staining; neurodegeneration; optical imaging; squalamine
DOI: 10.1096/fj.202201276R

Citations: 2
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