Genetically encoded chloride indicator with improved sensitivity

Markova, Olga; Mukhtarov, Marat; Réal, Éléonore; Jacob, Yves; Bregestovski, Pìotr

doi:10.1016/j.jneumeth.2007.12.016

Cited by 123 publications

(168 citation statements)

References 63 publications

Supporting

Mentioning

152

Contrasting

Order By: Relevance

“…A developmental decrease in [Cl 2 ] i has been well established in vertebrate neurons using both perforated patch electrophysiological recordings and genetically encoded fluorescent Cl 2 reporter proteins (Duebel et al 2006;Markova et al 2008). C. elegans membranes are not amenable to the perforated patch technique (Bellemer et al 2011), and our unpublished studies using fluorescent Cl 2 reporter proteins in C. elegans muscles have so far shown results too variable to make conclusions.…”

Section: Discussionmentioning

confidence: 93%

An Evolutionarily Conserved Switch in Response to GABA Affects Development and Behavior of the Locomotor Circuit of Caenorhabditis elegans

Han¹,

Bellemer

Koelle

2015

Genetics

View full text Add to dashboard Cite

The neurotransmitter gamma-aminobutyric acid (GABA) is depolarizing in the developing vertebrate brain, but in older animals switches to hyperpolarizing and becomes the major inhibitory neurotransmitter in adults. We discovered a similar developmental switch in GABA response in Caenorhabditis elegans and have genetically analyzed its mechanism and function in a well-defined circuit. Worm GABA neurons innervate body wall muscles to control locomotion. Activation of GABA A receptors with their agonist muscimol in newly hatched first larval (L1) stage animals excites muscle contraction and thus is depolarizing. At the mid-L1 stage, as the GABAergic neurons rewire onto their mature muscle targets, muscimol shifts to relaxing muscles and thus has switched to hyperpolarizing. This muscimol response switch depends on chloride transporters in the muscles analogous to those that control GABA response in mammalian neurons: the chloride accumulator sodium-potassium-chloride-cotransporter-1 (NKCC-1) is required for the early depolarizing muscimol response, while the two chloride extruders potassium-chloride-cotransporter-2 (KCC-2) and anion-bicarbonate-transporter-1 (ABTS-1) are required for the later hyperpolarizing response. Using mutations that disrupt GABA signaling, we found that neural circuit development still proceeds to completion but with an 6-hr delay. Using optogenetic activation of GABAergic neurons, we found that endogenous GABA A signaling in early L1 animals, although presumably depolarizing, does not cause an excitatory response. Thus a developmental depolarizing-tohyperpolarizing shift is an ancient conserved feature of GABA signaling, but existing theories for why this shift occurs appear inadequate to explain its function upon rigorous genetic analysis of a well-defined neural circuit.

show abstract

Section: Discussionmentioning

confidence: 93%

An Evolutionarily Conserved Switch in Response to GABA Affects Development and Behavior of the Locomotor Circuit of Caenorhabditis elegans

Han¹,

Bellemer

Koelle

2015

Genetics

View full text Add to dashboard Cite

show abstract

“…Most importantly, the polarity of input and output of the recorded neurons is unclear, because calcium is involved both in presynaptic and postsynaptic processes. Here the use of other tools, such as chloride sensitive reporters (Markova et al, 2008) or neurotransmitter release reporters (Miesenböck et al, 1998;Ng et al, 2002) would provide complementary data. Furthermore, because all measurements were performed on neuron populations, the polarity of individual neurons may be lost in our recordings.…”

Section: Discussionmentioning

confidence: 99%

Olfactory Information Processing in theDrosophilaAntennal Lobe: Anything Goes?

Silbering¹,

Okada²,

Ito³

et al. 2008

J. Neurosci.

119

131

View full text Add to dashboard Cite

When an animal smells an odor, olfactory sensory neurons generate an activity pattern across olfactory glomeruli of the first sensory neuropil, the insect antennal lobe or the vertebrate olfactory bulb. Here, several networks of local neurons interact with sensory neurons and with output neurons-insect projection neurons, or vertebrate mitral/tufted cells. The extent and form of information processing taking place in these local networks has been subject of controversy. To investigate the role of local neurons in odor information processing we have used the calcium sensor G-CaMP to perform in vivo recordings of odor-evoked spatiotemporal activity patterns in five genetically defined neuron populations of the antennal lobe of Drosophila melanogaster: three distinct populations of local neurons (two GABAergic and one cholinergic), as well as sensory neurons and projection neurons. Odor-specific and concentration dependent spatiotemporal response patterns varied among neuron populations. Activity transfer differed along the olfactory pathway for different glomerulus-odor combinations: we found cases of profile broadening and of linear and complex transfer. Moreover, the discriminability between the odors also varied across neuron populations and was maximal in projection neurons. Discriminatory power increased with higher odor concentrations over a wide dynamic range, but decreased at the highest concentration. These results show the complexity and diversity of odor information processing mechanisms across olfactory glomeruli in the fly antennal lobe.

show abstract

“…To test this hypothesis, we performed live-cell imaging experiments on biolistically transformed N. tabacum pollen tubes. We expressed the genetic anion-sensitive probe Chloride-Sensor to monitor changes in anion level (Markova et al, 2008). This cyan fluorescent protein (CFP)-yellow fluorescent protein (YFP) based FRET sensor for anions has a relatively high anion sensitivity (K app ;30 mM) in contrast with previous genetically encoded anion sensors (Lorenzen et al, 2004).…”

Section: Growing Pollen Tubes Possess a Cytosolic Anion Concentrationmentioning

confidence: 99%

Pollen Tube Growth Regulation by Free Anions Depends on the Interaction between the Anion Channel SLAH3 and Calcium-Dependent Protein Kinases CPK2 and CPK20

Gutermuth¹,

et al. 2013

View full text Add to dashboard Cite

(J.A.F.).Apical growth in pollen tubes (PTs) is associated with the presence of tip-focused ion gradients and fluxes, implying polar localization or regulation of the underlying transporters. The molecular identity and regulation of anion transporters in PTs is unknown. Here we report a negative gradient of cytosolic anion concentration focused on the tip, in negative correlation with the cytosolic Ca 2+ concentration. We hypothesized that a possible link between these two ions is based on the presence of Ca 2+ -dependent protein kinases (CPKs). We characterized anion channels and CPK transcripts in PTs and analyzed their localization. Yellow fluorescent protein (YFP) tagging of a homolog of SLOW ANION CHANNEL-ASSOCIATED1 (SLAH3:YFP) was widespread along PTs, but, in accordance with the anion efflux, CPK2/CPK20/CPK17/CPK34:YFP fluorescence was strictly localized at the tip plasma membrane. Expression of SLAH3 with either CPK2 or CPK20 (but not CPK17/CPK34) in Xenopus laevis oocytes elicited S-type anion channel currents. Interaction of SLAH3 with CPK2/CPK20 (but not CPK17/CPK34) was confirmed by Förster-resonance energy transfer fluorescence lifetime microscopy in Arabidopsis thaliana mesophyll protoplasts and bimolecular fluorescence complementation in living PTs. Compared with wild-type PTs, slah3-1 and slah3-2 as well as cpk2-1 cpk20-2 PTs had reduced anion currents. Double mutant cpk2-1 cpk20-2 and slah3-1 PTs had reduced extracellular anion fluxes at the tip. Our studies provide evidence for a Ca 2+ -dependent CPK2/CPK20 regulation of the anion channel SLAH3 to regulate PT growth.

show abstract

Genetically encoded chloride indicator with improved sensitivity

Cited by 123 publications

References 63 publications

An Evolutionarily Conserved Switch in Response to GABA Affects Development and Behavior of the Locomotor Circuit of Caenorhabditis elegans

An Evolutionarily Conserved Switch in Response to GABA Affects Development and Behavior of the Locomotor Circuit of Caenorhabditis elegans

Olfactory Information Processing in theDrosophilaAntennal Lobe: Anything Goes?

Pollen Tube Growth Regulation by Free Anions Depends on the Interaction between the Anion Channel SLAH3 and Calcium-Dependent Protein Kinases CPK2 and CPK20

Contact Info

Product

Resources

About