In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes

Abstract Background: The aminoglycoside antibiotic gentamicin is an ototoxic drug and has been used experimentally to investigate cochlear damage induced by noise. We have investigated the changes in the protein profile associated with caveolae in gentamicin treated and untreated spiral ligament (SL...

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Autores principales: Ghelfi, Elisa, Grondin, Yohann, Millet, Emil J., Bartos, Adam, Bortoni, Magda, Oliveira Gomes dos Santos, Clara, Treviño Villarreal, Humberto J., Sepúlveda Sepúlveda, Rosalinda, Rogers, Rick
Formato: Artículo
Lenguaje:inglés
Publicado: BioMed Central 2018
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Acceso en línea:http://eprints.uanl.mx/16314/1/159.pdf
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author Ghelfi, Elisa
Grondin, Yohann
Millet, Emil J.
Bartos, Adam
Bortoni, Magda
Oliveira Gomes dos Santos, Clara
Treviño Villarreal, Humberto J.
Sepúlveda Sepúlveda, Rosalinda
Rogers, Rick
author_facet Ghelfi, Elisa
Grondin, Yohann
Millet, Emil J.
Bartos, Adam
Bortoni, Magda
Oliveira Gomes dos Santos, Clara
Treviño Villarreal, Humberto J.
Sepúlveda Sepúlveda, Rosalinda
Rogers, Rick
author_sort Ghelfi, Elisa
collection Repositorio Institucional
description Abstract Background: The aminoglycoside antibiotic gentamicin is an ototoxic drug and has been used experimentally to investigate cochlear damage induced by noise. We have investigated the changes in the protein profile associated with caveolae in gentamicin treated and untreated spiral ligament (SL) pericytes, specialized cells in the blood labyrinth barrier of the inner ear microvasculature. Pericytes from various microvascular beds express caveolae, protein and cholesterol rich microdomains, which can undergo endocytosis and transcytosis to transport small molecules in and out the cells. A different protein profile in transport-specialized caveolae may induce pathological changes affecting the integrity of the blood labyrinth barrier and ultimately contributing to hearing loss. Method: Caveolae isolation from treated and untreated cells is achieved through ultracentrifugation of the lysates in discontinuous gradients. Mass spectrometry (LC-MS/MS) analysis identifies the proteins in the two groups. Proteins segregating with caveolae isolated from untreated SL pericytes are then compared to caveolae isolated from SL pericytes treated with the gentamicin for 24 h. Data are analyzed using bioinformatic tools. Results: The caveolae proteome in gentamicin treated cells shows that 40% of total proteins are uniquely associated with caveolae during the treatment, and 15% of the proteins normally associated with caveolae in untreated cell are suppressed. Bioinformatic analysis of the data shows a decreased expression of proteins involved in genetic information processing, and an increase in proteins involved in metabolism, vesicular transport and signal transduction in gentamicin treated cells. Several Rab GTPases proteins, ubiquitous transporters, uniquely segregate with caveolae and are significantly enriched in gentamicin treated cells. Conclusion: We report that gentamicin exposure modifies protein profile of caveolae from SL pericytes. We identified a pool of proteins which are uniquely segregating with caveolae during the treatment, mainly participating in metabolic and biosynthetic pathways, in transport pathways and in genetic information processing. Finally, we show for the first time proteins associated with caveolae SL pericytes linked to nonsyndromic hearing loss. Keywords: Ototoxic drug, Nonsyndromic hearing loss, Rab GTPase, GOrilla enrichment analysis, Proteomaps
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spelling eprints-163142020-06-01T12:59:09Z http://eprints.uanl.mx/16314/ In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes Ghelfi, Elisa Grondin, Yohann Millet, Emil J. Bartos, Adam Bortoni, Magda Oliveira Gomes dos Santos, Clara Treviño Villarreal, Humberto J. Sepúlveda Sepúlveda, Rosalinda Rogers, Rick QP Fisiología Abstract Background: The aminoglycoside antibiotic gentamicin is an ototoxic drug and has been used experimentally to investigate cochlear damage induced by noise. We have investigated the changes in the protein profile associated with caveolae in gentamicin treated and untreated spiral ligament (SL) pericytes, specialized cells in the blood labyrinth barrier of the inner ear microvasculature. Pericytes from various microvascular beds express caveolae, protein and cholesterol rich microdomains, which can undergo endocytosis and transcytosis to transport small molecules in and out the cells. A different protein profile in transport-specialized caveolae may induce pathological changes affecting the integrity of the blood labyrinth barrier and ultimately contributing to hearing loss. Method: Caveolae isolation from treated and untreated cells is achieved through ultracentrifugation of the lysates in discontinuous gradients. Mass spectrometry (LC-MS/MS) analysis identifies the proteins in the two groups. Proteins segregating with caveolae isolated from untreated SL pericytes are then compared to caveolae isolated from SL pericytes treated with the gentamicin for 24 h. Data are analyzed using bioinformatic tools. Results: The caveolae proteome in gentamicin treated cells shows that 40% of total proteins are uniquely associated with caveolae during the treatment, and 15% of the proteins normally associated with caveolae in untreated cell are suppressed. Bioinformatic analysis of the data shows a decreased expression of proteins involved in genetic information processing, and an increase in proteins involved in metabolism, vesicular transport and signal transduction in gentamicin treated cells. Several Rab GTPases proteins, ubiquitous transporters, uniquely segregate with caveolae and are significantly enriched in gentamicin treated cells. Conclusion: We report that gentamicin exposure modifies protein profile of caveolae from SL pericytes. We identified a pool of proteins which are uniquely segregating with caveolae during the treatment, mainly participating in metabolic and biosynthetic pathways, in transport pathways and in genetic information processing. Finally, we show for the first time proteins associated with caveolae SL pericytes linked to nonsyndromic hearing loss. Keywords: Ototoxic drug, Nonsyndromic hearing loss, Rab GTPase, GOrilla enrichment analysis, Proteomaps BioMed Central 2018-03-16 Article PeerReviewed text en cc_by_nc_nd http://eprints.uanl.mx/16314/1/159.pdf http://eprints.uanl.mx/16314/1.haspreviewThumbnailVersion/159.pdf Ghelfi, Elisa y Grondin, Yohann y Millet, Emil J. y Bartos, Adam y Bortoni, Magda y Oliveira Gomes dos Santos, Clara y Treviño Villarreal, Humberto J. y Sepúlveda Sepúlveda, Rosalinda y Rogers, Rick (2018) In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes. Proteome Science, 16 (1). pp. 1-26. ISSN 1477-5956 http://doi.org/10.1186/s12953-018-0132-x doi:10.1186/s12953-018-0132-x
spellingShingle QP Fisiología
Ghelfi, Elisa
Grondin, Yohann
Millet, Emil J.
Bartos, Adam
Bortoni, Magda
Oliveira Gomes dos Santos, Clara
Treviño Villarreal, Humberto J.
Sepúlveda Sepúlveda, Rosalinda
Rogers, Rick
In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes
thumbnail https://rediab.uanl.mx/themes/sandal5/images/online.png
title In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes
title_full In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes
title_fullStr In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes
title_full_unstemmed In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes
title_short In vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes
title_sort in vitro gentamicin exposure alters caveolae protein profile in cochlear spiral ligament pericytes
topic QP Fisiología
url http://eprints.uanl.mx/16314/1/159.pdf
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