Ludwig , A , Rivera Baeza , C & Uvarov , P 2017 , ' A noninvasive optical approach for assessing chloride extrusion activity of the K-Cl cotransporter KCC2 in neuronal cells ' , BMC Neuroscience , vol. 18 , 23 . https://doi.org/10.1186/s12868-017-0336-5
Title: | A noninvasive optical approach for assessing chloride extrusion activity of the K-Cl cotransporter KCC2 in neuronal cells |
Author: | Ludwig, Anastasia; Rivera Baeza, Claudio; Uvarov, Pavel |
Contributor organization: | Neuroscience Center University of Helsinki Helsinki In Vivo Animal Imaging Platform (HAIP) Claudio Rivera Baeza / Principal Investigator Medicum Department of Anatomy |
Date: | 2017-01-31 |
Language: | eng |
Number of pages: | 15 |
Belongs to series: | BMC Neuroscience |
ISSN: | 1471-2202 |
DOI: | https://doi.org/10.1186/s12868-017-0336-5 |
URI: | http://hdl.handle.net/10138/176706 |
Abstract: | Background: Cation-chloride cotransporters (CCCs) are indispensable for maintaining chloride homeostasis in multiple cell types, but K-Cl cotransporter KCC2 is the only CCC member with an exclusively neuronal expression in mammals. KCC2 is critical for rendering fast hyperpolarizing responses of ionotropic.-aminobutyric acid and glycine receptors in adult neurons, for neuronal migration in the developing central nervous system, and for the formation and maintenance of small dendritic protrusions-dendritic spines. Deficit in KCC2 expression and/or activity is associated with epilepsy and neuropathic pain, and effective strategies are required to search for novel drugs augmenting KCC2 function. Results: We revised current methods to develop a noninvasive optical approach for assessing KCC2 transport activity using a previously characterized genetically encoded chloride sensor. Our protocol directly assesses dynamics of KCC2-mediated chloride efflux and allows measuring genuine KCC2 activity with good spatial and temporal resolution. As a proof of concept, we used this approach to compare transport activities of the two known KCC2 splice isoforms, KCC2a and KCC2b, in mouse neuronal Neuro-2a cells. Conclusions: Our noninvasive optical protocol proved to be efficient for assessment of furosemide-sensitive chloride fluxes. Transport activities of the N-terminal splice isoforms KCC2a and KCC2b obtained by the novel approach matched to those reported previously using standard methods for measuring chloride fluxes. |
Subject: |
Genetically encoded chloride sensor
Slc12a5 gene KCC2 Inhibition GABA GREEN FLUORESCENT PROTEIN GENETICALLY-ENCODED CHLORIDE THROUGHPUT FUNCTIONAL ASSAY PRESYNAPTIC NERVE-TERMINALS INTRACELLULAR CHLORIDE HIPPOCAMPAL-NEURONS RAT-BRAIN SYNAPTIC INHIBITION POTASSIUM CHANNELS NEUROPATHIC PAIN 3112 Neurosciences 3124 Neurology and psychiatry |
Peer reviewed: | Yes |
Usage restriction: | openAccess |
Self-archived version: | publishedVersion |
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