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KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice
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| Veröffentlicht in: | Frontiers in cellular neuroscience 12(2018) Artikel-Nummer 7, 14 Seiten |
|---|---|
| Personen und Körperschaften: | , , , |
| Titel: | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice/ Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner |
| Format: | E-Book-Kapitel |
| Sprache: | Englisch |
| veröffentlicht: |
25 January 2018
|
| Gesamtaufnahme: |
: Frontiers in cellular neuroscience, 12(2018) Artikel-Nummer 7, 14 Seiten
, volume:12 |
| Schlagwörter: | |
| Quelle: | Verbunddaten SWB Lizenzfreie Online-Ressourcen |
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| 245 | 1 | 0 | |a KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice |c Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner |
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| 520 | |a Neuronal intracellular Cl- concentration ([Cl-]i) influences a wide range of processes such as neuronal inhibition, membrane potential dynamics, intracellular pH (pHi) or cell volume. Up to date, neuronal [Cl-]i has predominantly been studied in model systems of reduced complexity. Here, we implemented the genetically encoded ratiometric Cl- indicator Superclomeleon (SCLM) to estimate the steady-state [Cl-]i in cortical neurons from anesthetized and awake mice using 2-photon microscopy. Additionally, we implemented superecliptic pHluorin as a ratiometric sensor to estimate the intracellular steady-state pH (pHi) of mouse cortical neurons in vivo. We estimated an average resting [Cl-]i of 6 ± 2 mM with no evidence of subcellular gradients in the proximal somato-dendritic domain and an average somatic pHi of 7.1 ± 0.2. Neither [Cl-]i nor pHi were affected by isoflurane anesthesia. We deleted the cation-Cl- co-transporter KCC2 in single identified neurons of adult mice and found an increase of [Cl-]i to approximately 26 ± 8 mM, demonstrating that under in vivo conditions KCC2 produces low [Cl-]i in adult mouse neurons. In summary, neurons of the brain of awake adult mice exhibit a low and evenly distributed [Cl-]i in the proximal somato-dendritic compartment that is independent of anesthesia and requires KCC2 expression for its maintenance. | ||
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| author | Boffi, Juan C., Knabbe, Johannes, Kaiser, Michaela, Kuner, Thomas |
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| contents | Neuronal intracellular Cl- concentration ([Cl-]i) influences a wide range of processes such as neuronal inhibition, membrane potential dynamics, intracellular pH (pHi) or cell volume. Up to date, neuronal [Cl-]i has predominantly been studied in model systems of reduced complexity. Here, we implemented the genetically encoded ratiometric Cl- indicator Superclomeleon (SCLM) to estimate the steady-state [Cl-]i in cortical neurons from anesthetized and awake mice using 2-photon microscopy. Additionally, we implemented superecliptic pHluorin as a ratiometric sensor to estimate the intracellular steady-state pH (pHi) of mouse cortical neurons in vivo. We estimated an average resting [Cl-]i of 6 ± 2 mM with no evidence of subcellular gradients in the proximal somato-dendritic domain and an average somatic pHi of 7.1 ± 0.2. Neither [Cl-]i nor pHi were affected by isoflurane anesthesia. We deleted the cation-Cl- co-transporter KCC2 in single identified neurons of adult mice and found an increase of [Cl-]i to approximately 26 ± 8 mM, demonstrating that under in vivo conditions KCC2 produces low [Cl-]i in adult mouse neurons. In summary, neurons of the brain of awake adult mice exhibit a low and evenly distributed [Cl-]i in the proximal somato-dendritic compartment that is independent of anesthesia and requires KCC2 expression for its maintenance. |
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| spelling | Boffi, Juan C. VerfasserIn (DE-588)1156043832 (DE-627)1018601333 (DE-576)502001232 aut, KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner, 25 January 2018, Text txt rdacontent, Computermedien c rdamedia, Online-Ressource cr rdacarrier, Gesehen am 13.04.2018, Neuronal intracellular Cl- concentration ([Cl-]i) influences a wide range of processes such as neuronal inhibition, membrane potential dynamics, intracellular pH (pHi) or cell volume. Up to date, neuronal [Cl-]i has predominantly been studied in model systems of reduced complexity. Here, we implemented the genetically encoded ratiometric Cl- indicator Superclomeleon (SCLM) to estimate the steady-state [Cl-]i in cortical neurons from anesthetized and awake mice using 2-photon microscopy. Additionally, we implemented superecliptic pHluorin as a ratiometric sensor to estimate the intracellular steady-state pH (pHi) of mouse cortical neurons in vivo. We estimated an average resting [Cl-]i of 6 ± 2 mM with no evidence of subcellular gradients in the proximal somato-dendritic domain and an average somatic pHi of 7.1 ± 0.2. Neither [Cl-]i nor pHi were affected by isoflurane anesthesia. We deleted the cation-Cl- co-transporter KCC2 in single identified neurons of adult mice and found an increase of [Cl-]i to approximately 26 ± 8 mM, demonstrating that under in vivo conditions KCC2 produces low [Cl-]i in adult mouse neurons. In summary, neurons of the brain of awake adult mice exhibit a low and evenly distributed [Cl-]i in the proximal somato-dendritic compartment that is independent of anesthesia and requires KCC2 expression for its maintenance., chloride, in vivo, intracellular, neuronal, pH, ratiometric imaging., Knabbe, Johannes VerfasserIn (DE-588)1156044022 (DE-627)1018601619 (DE-576)502001534 aut, Kaiser, Michaela VerfasserIn (DE-588)1074365305 (DE-627)832065757 (DE-576)442610823 aut, Kuner, Thomas VerfasserIn (DE-588)1074267966 (DE-627)831945397 (DE-576)442234562 aut, Enthalten in Frontiers in cellular neuroscience Lausanne : Frontiers Research Foundation, 2007 12(2018) Artikel-Nummer 7, 14 Seiten Online-Ressource (DE-627)579826414 (DE-600)2452963-1 (DE-576)286502240 1662-5102 nnns, volume:12 year:2018, http://dx.doi.org/10.3389/fncel.2018.00007 Verlag Resolving-System kostenfrei Volltext, https://www.frontiersin.org/articles/10.3389/fncel.2018.00007/full Verlag kostenfrei Volltext, http://dx.doi.org/10.3389/fncel.2018.00007 LFER, LFER 2018-05-17T00:00:00Z |
| spellingShingle | Boffi, Juan C., Knabbe, Johannes, Kaiser, Michaela, Kuner, Thomas, KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice, Neuronal intracellular Cl- concentration ([Cl-]i) influences a wide range of processes such as neuronal inhibition, membrane potential dynamics, intracellular pH (pHi) or cell volume. Up to date, neuronal [Cl-]i has predominantly been studied in model systems of reduced complexity. Here, we implemented the genetically encoded ratiometric Cl- indicator Superclomeleon (SCLM) to estimate the steady-state [Cl-]i in cortical neurons from anesthetized and awake mice using 2-photon microscopy. Additionally, we implemented superecliptic pHluorin as a ratiometric sensor to estimate the intracellular steady-state pH (pHi) of mouse cortical neurons in vivo. We estimated an average resting [Cl-]i of 6 ± 2 mM with no evidence of subcellular gradients in the proximal somato-dendritic domain and an average somatic pHi of 7.1 ± 0.2. Neither [Cl-]i nor pHi were affected by isoflurane anesthesia. We deleted the cation-Cl- co-transporter KCC2 in single identified neurons of adult mice and found an increase of [Cl-]i to approximately 26 ± 8 mM, demonstrating that under in vivo conditions KCC2 produces low [Cl-]i in adult mouse neurons. In summary, neurons of the brain of awake adult mice exhibit a low and evenly distributed [Cl-]i in the proximal somato-dendritic compartment that is independent of anesthesia and requires KCC2 expression for its maintenance., chloride, in vivo, intracellular, neuronal, pH, ratiometric imaging. |
| swb_id_str | 50200553X |
| title | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice |
| title_auth | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice |
| title_full | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner |
| title_fullStr | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner |
| title_full_unstemmed | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner |
| title_in_hierarchy | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice / Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner, |
| title_short | KCC2-dependent steady-state intracellular chloride concentration and pH in cortical layer 2/3 neurons of anesthetized and awake mice |
| title_sort | kcc2 dependent steady state intracellular chloride concentration and ph in cortical layer 2 3 neurons of anesthetized and awake mice |
| topic | chloride, in vivo, intracellular, neuronal, pH, ratiometric imaging. |
| topic_facet | chloride, in vivo, intracellular, neuronal, pH, ratiometric imaging. |
| url | http://dx.doi.org/10.3389/fncel.2018.00007, https://www.frontiersin.org/articles/10.3389/fncel.2018.00007/full |