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Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes

Gut–brain axis (GBA) communication relies on serotonin (5-HT) signaling between the gut epithelium and the peripheral nervous system, where 5-HT release patterns from the basolateral (i.e., bottom) side of the epithelium activate nerve afferents. There have been few quantitative studies of this gut-...

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Autores principales: Chapin, Ashley A., Rajasekaran, Pradeep R., Quan, David N., Hu, Liangbing, Herberholz, Jens, Bentley, William E., Ghodssi, Reza
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433419/
https://www.ncbi.nlm.nih.gov/pubmed/34567700
http://dx.doi.org/10.1038/s41378-020-00184-4
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author Chapin, Ashley A.
Rajasekaran, Pradeep R.
Quan, David N.
Hu, Liangbing
Herberholz, Jens
Bentley, William E.
Ghodssi, Reza
author_facet Chapin, Ashley A.
Rajasekaran, Pradeep R.
Quan, David N.
Hu, Liangbing
Herberholz, Jens
Bentley, William E.
Ghodssi, Reza
author_sort Chapin, Ashley A.
collection PubMed
description Gut–brain axis (GBA) communication relies on serotonin (5-HT) signaling between the gut epithelium and the peripheral nervous system, where 5-HT release patterns from the basolateral (i.e., bottom) side of the epithelium activate nerve afferents. There have been few quantitative studies of this gut-neuron signaling due to a lack of real-time measurement tools that can access the basolateral gut epithelium. In vitro platforms allow quantitative studies of cultured gut tissue, but they mainly employ offline and endpoint assays that cannot resolve dynamic molecular-release patterns. Here, we present the modification of a microporous cell culture membrane with carbon nanotube-coated gold (Au-CNT) electrodes capable of continuous, label-free, and direct detection of 5-HT at physiological concentrations. Electrochemical characterization of single-walled carbon nanotube (SWCNT)-coated Au electrodes shows increased electroactive surface area, 5-HT specificity, sensitivity, and saturation time, which are correlated with the CNT film drop-cast volume. Two microliters of CNT films, with a 10-min saturation time, 0.6 μA/μM 5-HT sensitivity, and reliable detection within a linear range of 500 nM–10 μM 5-HT, can be targeted for high-concentration, high-time-resolution 5-HT monitoring. CNT films (12.5 μL) with a 2-h saturation time, 4.5 μA/μM 5-HT sensitivity, and quantitative detection in the linear range of 100 nM–1 μM can target low concentrations with low time resolution. These electrodes achieved continuous detection of dynamic diffusion across the porous membrane, mimicking basolateral 5-HT release from cells, and detection of cell-released 5-HT from separately cultured RIN14B cell supernatant. Electrode-integrated cell culture systems such as this can improve in vitro molecular detection mechanisms and aid in quantitative GBA signaling studies.
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spelling pubmed-84334192021-09-24 Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes Chapin, Ashley A. Rajasekaran, Pradeep R. Quan, David N. Hu, Liangbing Herberholz, Jens Bentley, William E. Ghodssi, Reza Microsyst Nanoeng Article Gut–brain axis (GBA) communication relies on serotonin (5-HT) signaling between the gut epithelium and the peripheral nervous system, where 5-HT release patterns from the basolateral (i.e., bottom) side of the epithelium activate nerve afferents. There have been few quantitative studies of this gut-neuron signaling due to a lack of real-time measurement tools that can access the basolateral gut epithelium. In vitro platforms allow quantitative studies of cultured gut tissue, but they mainly employ offline and endpoint assays that cannot resolve dynamic molecular-release patterns. Here, we present the modification of a microporous cell culture membrane with carbon nanotube-coated gold (Au-CNT) electrodes capable of continuous, label-free, and direct detection of 5-HT at physiological concentrations. Electrochemical characterization of single-walled carbon nanotube (SWCNT)-coated Au electrodes shows increased electroactive surface area, 5-HT specificity, sensitivity, and saturation time, which are correlated with the CNT film drop-cast volume. Two microliters of CNT films, with a 10-min saturation time, 0.6 μA/μM 5-HT sensitivity, and reliable detection within a linear range of 500 nM–10 μM 5-HT, can be targeted for high-concentration, high-time-resolution 5-HT monitoring. CNT films (12.5 μL) with a 2-h saturation time, 4.5 μA/μM 5-HT sensitivity, and quantitative detection in the linear range of 100 nM–1 μM can target low concentrations with low time resolution. These electrodes achieved continuous detection of dynamic diffusion across the porous membrane, mimicking basolateral 5-HT release from cells, and detection of cell-released 5-HT from separately cultured RIN14B cell supernatant. Electrode-integrated cell culture systems such as this can improve in vitro molecular detection mechanisms and aid in quantitative GBA signaling studies. Nature Publishing Group UK 2020-09-07 /pmc/articles/PMC8433419/ /pubmed/34567700 http://dx.doi.org/10.1038/s41378-020-00184-4 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Chapin, Ashley A.
Rajasekaran, Pradeep R.
Quan, David N.
Hu, Liangbing
Herberholz, Jens
Bentley, William E.
Ghodssi, Reza
Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes
title Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes
title_full Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes
title_fullStr Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes
title_full_unstemmed Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes
title_short Electrochemical measurement of serotonin by Au-CNT electrodes fabricated on microporous cell culture membranes
title_sort electrochemical measurement of serotonin by au-cnt electrodes fabricated on microporous cell culture membranes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433419/
https://www.ncbi.nlm.nih.gov/pubmed/34567700
http://dx.doi.org/10.1038/s41378-020-00184-4
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