ArticleFrontiers in molecular neuroscience2013
Improved method for efficient imaging of intracellular Cl(-) with Cl-Sensor using conventional fluorescence setup.
Article in Frontiers in molecular neuroscience, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 30 citations in OpenAlex.
- Uncovering novel KCC2 regulatory motifs through a comprehensive transposon-based mutant library.Frontiers in molecular neuroscience · 2024Article
- KFrontiers in cellular neuroscience · 2023Article
- Long-Term Imaging Reveals a Circadian Rhythm of Intracellular Chloride in Neurons of the Suprachiasmatic Nucleus.Journal of biological rhythms · 2022Article
- Smoothened receptor signaling regulates the developmental shift of GABA polarity in rat somatosensory cortex.Journal of cell science · 2020Article
- Distal convoluted tubule ClAmerican journal of physiology. Renal physiology · 2020Article
- Impaired regulation of KCC2 phosphorylation leads to neuronal network dysfunction and neurodevelopmental pathology.Science signaling · 2019Article
- Effects of salt-loading on supraoptic vasopressin neurones assessed by ClopHensorN chloride imaging.Journal of neuroendocrinology · 2019Article
- The small molecule CLP257 does not modify activity of the KNature medicine · 2017Article
- Molecular architecture of potassium chloride co-transporter KCC2.Scientific reports · 2017Article
- Intracellular Chloride Regulation in AVP+ and VIP+ Neurons of the Suprachiasmatic Nucleus.Scientific reports · 2017Article
- A noninvasive optical approach for assessing chloride extrusion activity of the K-Cl cotransporter KCC2 in neuronal cells.BMC neuroscience · 2017Article
- Molecular cloning and biochemical characterization of two cation chloride cotransporter subfamily members of Hydra vulgaris.PloS one · 2017Article
- Optogenetic Visualization of Presynaptic Tonic Inhibition of Cerebellar Parallel Fibers.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2016Article
- The K(+)-Cl(-) Cotransporter KCC2 and Chloride Homeostasis: Potential Therapeutic Target in Acute Central Nervous System Injury.Molecular neurobiology · 2016Review
- Selective inhibition of KCC2 leads to hyperexcitability and epileptiform discharges in hippocampal slices and in vivo.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2015Article
- Genetically encoded impairment of neuronal KCC2 cotransporter function in human idiopathic generalized epilepsy.EMBO reports · 2014Article
- Current view on the functional regulation of the neuronal K(+)-Cl(-) cotransporter KCC2.Frontiers in cellular neuroscience · 2014Review
- Transgenic mouse lines for non-invasive ratiometric monitoring of intracellular chloride.Frontiers in molecular neuroscience · 2013Article
- Molecular probes and switches for functional analysis of receptors, ion channels and synaptic networks.Frontiers in molecular neuroscience · 2013Article
- Calibration and functional analysis of three genetically encoded Cl(-)/pH sensors.Frontiers in molecular neuroscience · 2013Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Chloride (Cl(-)) homeostasis is known to be fundamental for central nervous system functioning. Alterations in intracellular Cl(-) concentration ([Cl(-)]i) and changes in the efficacy of Cl(-) extrusion are involved in numerous neurological disorders. Therefore, there is a strong need for studies of the dynamics of [Cl(-)]i in different cell types under physiological conditions and during pathology. Several previous works reported having successfully achieved recording of [Cl(-)]i using genetically encoded Cl-Sensor that is composed of the cyan fluorescent protein (CFP) and Cl(-)-sensitive mutant of the yellow fluorescent protein (YFPCl). However, all reported works were performed using specially designed setups with ultra-sensitive CCD cameras. Our multiple attempts to monitor Cl(-)-dependent fluorescence of Cl-Sensor using conventional epifluorescence microscopes did not yield successful results. In the present work, we have analysed the reason of our failures and found that they were caused by a strong inactivation of the YFPCl component of Cl-Sensor during excitation of the CFP with 430 nm light. Based on the obtained results, we reduced 20-fold the intensity of the 430 nm excitation and modified the recording protocol that allows now stable long-lasting ratiometric measurements of Cl-Sensor fluorescence in different cell types including cultured hippocampal neurons and their tiny dendrites and spines. Simultaneous imaging and patch clamp recording revealed that in mature neurons, the novel protocol allows detection of as little as 2 mM changes of [Cl(-)]i from the resting level of 5-10 mM. We demonstrate also a usefulness of the developed [Cl(-)]i measurement procedure for large scale screening of the activity of exogenously expressed potassium-chloride co-transporter KCC2, a major neuronal Cl(-) extruder that is implicated in numerous neurological disorders and is a target for novel therapeutical treatments.
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