Genetically encoded Cl-Sensor as a tool for monitoring of Cl-dependent processes in small neuronal compartments

Waseem, Tatyana V.; Mukhtarov, Marat; Buldakova, Svetlana; Medina, Igor; Bregestovski, Pìotr

doi:10.1016/j.jneumeth.2010.08.002

Cited by 62 publications

(81 citation statements)

References 72 publications

(70 reference statements)

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“…Cell Culture Preparation and Electrophysiological Recording-Chinese hamster ovary (CHO-K1) cells were obtained from the American Type Tissue Culture Collection (ATCC, Molsheim, France) and maintained as described previously (18). One day before the transfection, cells were plated on the coverslips (12-14 mm in diameter), which were placed inside 35-mm cell culture dishes with 2 ml of medium.…”

Section: Methodsmentioning

confidence: 99%

Water-soluble LYNX1 Residues Important for Interaction with Muscle-type and/or Neuronal Nicotinic Receptors

Lyukmanova

Shulepko

Buldakova

et al. 2013

Journal of Biological Chemistry

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Background: Information was not available about prototoxin LYNX1 amino acid residues involved in binding to muscle and neuronal nicotinic receptors. Results: A series of water-soluble LYNX1 (ws-LYNX1) mutants was obtained and their interaction with nicotinic receptors was analyzed. Conclusion: There are both common and selective ws-LYNX1 residues recognizing distinct receptor types. Significance: For the first time, several functionally important residues in ws-LYNX1 are identified.

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Section: Methodsmentioning

confidence: 99%

Water-soluble LYNX1 Residues Important for Interaction with Muscle-type and/or Neuronal Nicotinic Receptors

Lyukmanova

Shulepko

Buldakova

et al. 2013

Journal of Biological Chemistry

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“…Recent in vitro studies demonstrated the existence of spatial differences in the values of E GABA and DF GABA (REFS 183,184) that are attributable to subcellular heterogeneity of CCC localization 185,186 . In neocortical and hippocampal principal neurons, E GABA and DF GABA shift towards more hyperpolarizing values from the AIS to the soma as a result of high NKCC1 levels and negligible levels of KCC2 in the AIS 183,185–187 (but see REF.…”

Section: Cccs Regulate Inhibitory Signallingmentioning

confidence: 99%

Cation-chloride cotransporters in neuronal development, plasticity and disease

et al. 2014

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Electrical activity in neurons requires a seamless functional coupling between plasmalemmal ion channels and ion transporters. Although ion channels have been studied intensively for several decades, research on ion transporters is in its infancy. In recent years, it has become evident that one family of ion transporters, cation-chloride cotransporters (CCCs), and in particular K+–Cl− cotransporter 2 (KCC2), have seminal roles in shaping GABAergic signalling and neuronal connectivity. Studying the functions of these transporters may lead to major paradigm shifts in our understanding of the mechanisms underlying brain development and plasticity in health and disease.

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“…The addition of some other ionophores has been reported; e.g., monensin and ionomycin for clamping intracellular milieu at fixed values of sodium concentration (Harootunian et al, 1989) and the protonophore CCCP for controlling pH (Kneen et al, 1998). Alternative methods for in situ calibration in cell cultures were proposed by permeabilizing the cell membrane with digitonin (Kneen et al, 1998) or natural surfactants like the triterpenoid saponin, β-escin (Black et al, 2004; Nausch et al, 2008; Waseem et al, 2010). Although calibration procedures using β-escin and the ionophores nigericin and trybutyltin have been reported to yield encouragingly equivalent results (Watts et al, 2012), it is likely that the many variables potentially affecting in situ calibration will produce less precise estimates of the intrinsic molecular parameters when compared to spectroscopy measurements performed in a cuvette.…”

Section: Calibrationmentioning

confidence: 99%

Twenty years of fluorescence imaging of intracellular chloride

Arosio

Ratto

2014

Front. Cell. Neurosci.

111

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Chloride homeostasis has a pivotal role in controlling neuronal excitability in the adult brain and during development. The intracellular concentration of chloride is regulated by the dynamic equilibrium between passive fluxes through membrane conductances and the active transport mediated by importers and exporters. In cortical neurons, chloride fluxes are coupled to network activity by the opening of the ionotropic GABAA receptors that provides a direct link between the activity of interneurons and chloride fluxes. These molecular mechanisms are not evenly distributed and regulated over the neuron surface and this fact can lead to a compartmentalized control of the intracellular concentration of chloride. The inhibitory drive provided by the activity of the GABAA receptors depends on the direction and strength of the associated currents, which are ultimately dictated by the gradient of chloride, the main charge carrier flowing through the GABAA channel. Thus, the intracellular distribution of chloride determines the local strength of ionotropic inhibition and influences the interaction between converging excitation and inhibition. The importance of chloride regulation is also underlined by its involvement in several brain pathologies, including epilepsy and disorders of the autistic spectra. The full comprehension of the physiological meaning of GABAergic activity on neurons requires the measurement of the spatiotemporal dynamics of chloride fluxes across the membrane. Nowadays, there are several available tools for the task, and both synthetic and genetically encoded indicators have been successfully used for chloride imaging. Here, we will review the available sensors analyzing their properties and outlining desirable future developments.

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Genetically encoded Cl-Sensor as a tool for monitoring of Cl-dependent processes in small neuronal compartments

Cited by 62 publications

References 72 publications

Water-soluble LYNX1 Residues Important for Interaction with Muscle-type and/or Neuronal Nicotinic Receptors

Water-soluble LYNX1 Residues Important for Interaction with Muscle-type and/or Neuronal Nicotinic Receptors

Cation-chloride cotransporters in neuronal development, plasticity and disease

Twenty years of fluorescence imaging of intracellular chloride

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