Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of Ascorbate

The impaired blood flow to the brain causes a decrease in the supply of oxygen that can result in cerebral ischemia; if the blood flow is not restored quickly, neuronal injury or death will occur. Under hypoxic conditions, the production of nitric oxide (<sup>●</sup>NO), via the classica...

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Main Authors: Tiago Monteiro, Cândida Dias, Cátia F. Lourenço, Ana Ledo, Rui M. Barbosa, M. Gabriela Almeida
Format: Article
Language:English
Published: MDPI AG 2021-08-01
Series:Biosensors
Subjects:
Online Access:https://www.mdpi.com/2079-6374/11/8/277
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author Tiago Monteiro
Cândida Dias
Cátia F. Lourenço
Ana Ledo
Rui M. Barbosa
M. Gabriela Almeida
author_facet Tiago Monteiro
Cândida Dias
Cátia F. Lourenço
Ana Ledo
Rui M. Barbosa
M. Gabriela Almeida
author_sort Tiago Monteiro
collection DOAJ
description The impaired blood flow to the brain causes a decrease in the supply of oxygen that can result in cerebral ischemia; if the blood flow is not restored quickly, neuronal injury or death will occur. Under hypoxic conditions, the production of nitric oxide (<sup>●</sup>NO), via the classical L-arginine–<sup>●</sup>NO synthase pathway, is reduced, which can compromise <sup>●</sup>NO-dependent vasodilation. However, the alternative nitrite (NO<sub>2</sub><sup>−</sup>) reduction to <sup>●</sup>NO, under neuronal hypoxia and ischemia conditions, has been viewed as an in vivo storage pool of <sup>●</sup>NO, complementing its enzymatic synthesis. Brain research is thus demanding suitable tools to probe nitrite’s temporal and spatial dynamics in vivo. In this work, we propose a new method for the real-time measurement of nitrite concentration in the brain extracellular space, using fast-scan cyclic voltammetry (FSCV) and carbon microfiber electrodes as sensing probes. In this way, nitrite was detected anodically and in vitro, in the 5–500 µM range, in the presence of increasing physiological concentrations of ascorbate (100–500 µM). These sensors were then tested for real-time and in vivo recordings in the anesthetized rat hippocampus; using fast electrochemical techniques, local and reproducible transients of nitrite oxidation signals were observed, upon pressure ejection of an exogenous nitrite solution into the brain tissue. Nitrite microsensors are thus a valuable tool for investigating the role of this inorganic anion in brain redox signaling.
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spelling doaj.art-c4e70ae8d6f748a48ef68b8415d5495b2023-11-22T06:57:44ZengMDPI AGBiosensors2079-63742021-08-0111827710.3390/bios11080277Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of AscorbateTiago Monteiro0Cândida Dias1Cátia F. Lourenço2Ana Ledo3Rui M. Barbosa4M. Gabriela Almeida5UCIBIO—Applied Molecular Biosciences Unit, REQUIMTE—Rede de Química e Tecnologia, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Monte de Caparica, PortugalCenter for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, PortugalCenter for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, PortugalCenter for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, PortugalCenter for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, PortugalUCIBIO—Applied Molecular Biosciences Unit, REQUIMTE—Rede de Química e Tecnologia, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Monte de Caparica, PortugalThe impaired blood flow to the brain causes a decrease in the supply of oxygen that can result in cerebral ischemia; if the blood flow is not restored quickly, neuronal injury or death will occur. Under hypoxic conditions, the production of nitric oxide (<sup>●</sup>NO), via the classical L-arginine–<sup>●</sup>NO synthase pathway, is reduced, which can compromise <sup>●</sup>NO-dependent vasodilation. However, the alternative nitrite (NO<sub>2</sub><sup>−</sup>) reduction to <sup>●</sup>NO, under neuronal hypoxia and ischemia conditions, has been viewed as an in vivo storage pool of <sup>●</sup>NO, complementing its enzymatic synthesis. Brain research is thus demanding suitable tools to probe nitrite’s temporal and spatial dynamics in vivo. In this work, we propose a new method for the real-time measurement of nitrite concentration in the brain extracellular space, using fast-scan cyclic voltammetry (FSCV) and carbon microfiber electrodes as sensing probes. In this way, nitrite was detected anodically and in vitro, in the 5–500 µM range, in the presence of increasing physiological concentrations of ascorbate (100–500 µM). These sensors were then tested for real-time and in vivo recordings in the anesthetized rat hippocampus; using fast electrochemical techniques, local and reproducible transients of nitrite oxidation signals were observed, upon pressure ejection of an exogenous nitrite solution into the brain tissue. Nitrite microsensors are thus a valuable tool for investigating the role of this inorganic anion in brain redox signaling.https://www.mdpi.com/2079-6374/11/8/277carbon fiber microelectrodesfast-scan cyclic voltammetrynitritehippocampus
spellingShingle Tiago Monteiro
Cândida Dias
Cátia F. Lourenço
Ana Ledo
Rui M. Barbosa
M. Gabriela Almeida
Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of Ascorbate
Biosensors
carbon fiber microelectrodes
fast-scan cyclic voltammetry
nitrite
hippocampus
title Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of Ascorbate
title_full Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of Ascorbate
title_fullStr Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of Ascorbate
title_full_unstemmed Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of Ascorbate
title_short Microelectrode Sensor for Real-Time Measurements of Nitrite in the Living Brain, in the Presence of Ascorbate
title_sort microelectrode sensor for real time measurements of nitrite in the living brain in the presence of ascorbate
topic carbon fiber microelectrodes
fast-scan cyclic voltammetry
nitrite
hippocampus
url https://www.mdpi.com/2079-6374/11/8/277
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