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 implemen...
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| Main Authors: | , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
25 January 2018
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| In: |
Frontiers in cellular neuroscience
Year: 2018, Volume: 12 |
| ISSN: | 1662-5102 |
| DOI: | 10.3389/fncel.2018.00007 |
| Online Access: | Verlag, kostenfrei, Volltext: http://dx.doi.org/10.3389/fncel.2018.00007 Verlag, kostenfrei, Volltext: https://www.frontiersin.org/articles/10.3389/fncel.2018.00007/full 
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| Author Notes: | Juan C. Boffi, Johannes Knabbe, Michaela Kaiser and Thomas Kuner |
| Summary: | 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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| Item Description: | Gesehen am 13.04.2018 |
| Physical Description: | Online Resource |
| ISSN: | 1662-5102 |
| DOI: | 10.3389/fncel.2018.00007 |