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Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives

The styryl dye FM1-43 is widely used to study endocytosis but behaves as a permeant blocker of the mechano-electrical transducer (MET) channel in sensory hair cells, loading rapidly and specifically into the cytoplasm of hair cells in a MET channel-dependent manner. Patch clamp recordings of mouse o...

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Autores principales: Derudas, Marco, O’Reilly, Molly, Kirkwood, Nerissa K., Kenyon, Emma J., Grimsey, Sybil, Kitcher, Siân R., Workman, Shawna, Bull, James C., Ward, Simon E., Kros, Corné J., Richardson, Guy P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10601989/
https://www.ncbi.nlm.nih.gov/pubmed/37900280
http://dx.doi.org/10.3389/fcell.2023.1247324
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author Derudas, Marco
O’Reilly, Molly
Kirkwood, Nerissa K.
Kenyon, Emma J.
Grimsey, Sybil
Kitcher, Siân R.
Workman, Shawna
Bull, James C.
Ward, Simon E.
Kros, Corné J.
Richardson, Guy P.
author_facet Derudas, Marco
O’Reilly, Molly
Kirkwood, Nerissa K.
Kenyon, Emma J.
Grimsey, Sybil
Kitcher, Siân R.
Workman, Shawna
Bull, James C.
Ward, Simon E.
Kros, Corné J.
Richardson, Guy P.
author_sort Derudas, Marco
collection PubMed
description The styryl dye FM1-43 is widely used to study endocytosis but behaves as a permeant blocker of the mechano-electrical transducer (MET) channel in sensory hair cells, loading rapidly and specifically into the cytoplasm of hair cells in a MET channel-dependent manner. Patch clamp recordings of mouse outer hair cells (OHCs) were used to determine how a series of structural modifications of FM1-43 affect MET channel block. Fluorescence microscopy was used to assess how the modifications influence hair-cell loading in mouse cochlear cultures and zebrafish neuromasts. Cochlear cultures were also used to evaluate otoprotective potential of the modified FM1-43 derivatives. Structure-activity relationships reveal that the lipophilic tail and the cationic head group of FM1-43 are both required for MET channel block in mouse cochlear OHCs; neither moiety alone is sufficient. The extent of MET channel block is augmented by increasing the lipophilicity/bulkiness of the tail, by reducing the number of positive charges in the head group from two to one, or by increasing the distance between the two charged head groups. Loading assays with zebrafish neuromasts and mouse cochlear cultures are broadly in accordance with these observations but reveal a loss of hair-cell specific labelling with increasing lipophilicity. Although FM1-43 and many of its derivatives are generally cytotoxic when tested on cochlear cultures in the presence of an equimolar concentration of the ototoxic antibiotic gentamicin (5 µM), at a 10-fold lower concentration (0.5 µM), two of the derivatives protect OHCs from cell death caused by 48 h-exposure to 5 µM gentamicin.
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spelling pubmed-106019892023-10-27 Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives Derudas, Marco O’Reilly, Molly Kirkwood, Nerissa K. Kenyon, Emma J. Grimsey, Sybil Kitcher, Siân R. Workman, Shawna Bull, James C. Ward, Simon E. Kros, Corné J. Richardson, Guy P. Front Cell Dev Biol Cell and Developmental Biology The styryl dye FM1-43 is widely used to study endocytosis but behaves as a permeant blocker of the mechano-electrical transducer (MET) channel in sensory hair cells, loading rapidly and specifically into the cytoplasm of hair cells in a MET channel-dependent manner. Patch clamp recordings of mouse outer hair cells (OHCs) were used to determine how a series of structural modifications of FM1-43 affect MET channel block. Fluorescence microscopy was used to assess how the modifications influence hair-cell loading in mouse cochlear cultures and zebrafish neuromasts. Cochlear cultures were also used to evaluate otoprotective potential of the modified FM1-43 derivatives. Structure-activity relationships reveal that the lipophilic tail and the cationic head group of FM1-43 are both required for MET channel block in mouse cochlear OHCs; neither moiety alone is sufficient. The extent of MET channel block is augmented by increasing the lipophilicity/bulkiness of the tail, by reducing the number of positive charges in the head group from two to one, or by increasing the distance between the two charged head groups. Loading assays with zebrafish neuromasts and mouse cochlear cultures are broadly in accordance with these observations but reveal a loss of hair-cell specific labelling with increasing lipophilicity. Although FM1-43 and many of its derivatives are generally cytotoxic when tested on cochlear cultures in the presence of an equimolar concentration of the ototoxic antibiotic gentamicin (5 µM), at a 10-fold lower concentration (0.5 µM), two of the derivatives protect OHCs from cell death caused by 48 h-exposure to 5 µM gentamicin. Frontiers Media S.A. 2023-10-10 /pmc/articles/PMC10601989/ /pubmed/37900280 http://dx.doi.org/10.3389/fcell.2023.1247324 Text en Copyright © 2023 Derudas, O’Reilly, Kirkwood, Kenyon, Grimsey, Kitcher, Workman, Bull, Ward, Kros and Richardson. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Derudas, Marco
O’Reilly, Molly
Kirkwood, Nerissa K.
Kenyon, Emma J.
Grimsey, Sybil
Kitcher, Siân R.
Workman, Shawna
Bull, James C.
Ward, Simon E.
Kros, Corné J.
Richardson, Guy P.
Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives
title Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives
title_full Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives
title_fullStr Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives
title_full_unstemmed Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives
title_short Charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by FM1-43 and its derivatives
title_sort charge and lipophilicity are required for effective block of the hair-cell mechano-electrical transducer channel by fm1-43 and its derivatives
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10601989/
https://www.ncbi.nlm.nih.gov/pubmed/37900280
http://dx.doi.org/10.3389/fcell.2023.1247324
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