Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices
Summary: Reactive oxygen species (ROS) are important signaling molecules for physiologic processes such as acute response to hypoxia. However, reliable real-time ROS measurement in cells has been a long-standing methodological challenge. Here, we present a protocol to record acute changes in ROS pro...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-06-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721002422 |
| _version_ | 1818926419084836864 |
|---|---|
| author | Lin Gao Ignacio Arias-Mayenco Patricia Ortega-Sáenz José López-Barneo |
| author_facet | Lin Gao Ignacio Arias-Mayenco Patricia Ortega-Sáenz José López-Barneo |
| author_sort | Lin Gao |
| collection | DOAJ |
| description | Summary: Reactive oxygen species (ROS) are important signaling molecules for physiologic processes such as acute response to hypoxia. However, reliable real-time ROS measurement in cells has been a long-standing methodological challenge. Here, we present a protocol to record acute changes in ROS production in sensory cells from mouse carotid body (CB) slices using redox-sensitive green fluorescent protein probes and microfluorimetry. This protocol provides sensitive and reproducible quantification of ROS during acute hypoxia in different subcellular compartments of CB glomus cells.For complete details on the use and execution of this protocol, please refer to Fernández-Agüera et al. (2015) and Arias-Mayenco et al. (2018). |
| first_indexed | 2024-12-20T02:56:49Z |
| format | Article |
| id | doaj.art-19197bd2ef5049bc970974d2b14c4e1f |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-20T02:56:49Z |
| publishDate | 2021-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-19197bd2ef5049bc970974d2b14c4e1f2022-12-21T19:55:54ZengElsevierSTAR Protocols2666-16672021-06-0122100535Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slicesLin Gao0Ignacio Arias-Mayenco1Patricia Ortega-Sáenz2José López-Barneo3Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, 41013 Seville, Spain; Departamento de Fisiología Médica y Biofísica, Facultad de Medicina, Universidad de Sevilla, 41013 Seville, Spain; Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), 28031 Madrid, SpainInstituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, 41013 Seville, SpainInstituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, 41013 Seville, Spain; Departamento de Fisiología Médica y Biofísica, Facultad de Medicina, Universidad de Sevilla, 41013 Seville, Spain; Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), 28031 Madrid, SpainInstituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, 41013 Seville, Spain; Departamento de Fisiología Médica y Biofísica, Facultad de Medicina, Universidad de Sevilla, 41013 Seville, Spain; Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), 28031 Madrid, Spain; Corresponding authorSummary: Reactive oxygen species (ROS) are important signaling molecules for physiologic processes such as acute response to hypoxia. However, reliable real-time ROS measurement in cells has been a long-standing methodological challenge. Here, we present a protocol to record acute changes in ROS production in sensory cells from mouse carotid body (CB) slices using redox-sensitive green fluorescent protein probes and microfluorimetry. This protocol provides sensitive and reproducible quantification of ROS during acute hypoxia in different subcellular compartments of CB glomus cells.For complete details on the use and execution of this protocol, please refer to Fernández-Agüera et al. (2015) and Arias-Mayenco et al. (2018).http://www.sciencedirect.com/science/article/pii/S2666166721002422Cell BiologyMetabolismMicroscopyMolecular/Chemical Probes |
| spellingShingle | Lin Gao Ignacio Arias-Mayenco Patricia Ortega-Sáenz José López-Barneo Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices STAR Protocols Cell Biology Metabolism Microscopy Molecular/Chemical Probes |
| title | Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices |
| title_full | Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices |
| title_fullStr | Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices |
| title_full_unstemmed | Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices |
| title_short | Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices |
| title_sort | using redox sensitive fluorescent probes to record real time reactive oxygen species production in cells from mouse carotid body slices |
| topic | Cell Biology Metabolism Microscopy Molecular/Chemical Probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721002422 |
| work_keys_str_mv | AT lingao usingredoxsensitivefluorescentprobestorecordrealtimereactiveoxygenspeciesproductionincellsfrommousecarotidbodyslices AT ignacioariasmayenco usingredoxsensitivefluorescentprobestorecordrealtimereactiveoxygenspeciesproductionincellsfrommousecarotidbodyslices AT patriciaortegasaenz usingredoxsensitivefluorescentprobestorecordrealtimereactiveoxygenspeciesproductionincellsfrommousecarotidbodyslices AT joselopezbarneo usingredoxsensitivefluorescentprobestorecordrealtimereactiveoxygenspeciesproductionincellsfrommousecarotidbodyslices |