Muscarinic depolarization of SH-SY5Y human neuroblastoma cells as determined using oxonol V

Kukkonen, Jyrki P.; Hautala, Rasmus; Åkerman, Karl E.O.

doi:10.1016/0304-3940(96)12781-6

Cited by 5 publications

(4 citation statements)

References 24 publications

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“…We propose a fast fluorimetric test based on nonintrusive measurement of changes in membrane potential in neuroblastoma cells. The applicability of the optical probe oxonol to measurements of membrane potential in human neuroblastoma cells has already been demonstrated (24). In this paper we have investigated the effectiveness of the optical probe bis(1,3-diethylthiobarbituric acid)trimethine oxonol for detection and quantitation of PSP toxins.…”

Section: Discussionmentioning

confidence: 98%

A Fluorimetric Method Based on Changes in Membrane Potential for Screening Paralytic Shellfish Toxins in Mussels

Louzao

Vieytes

Sousa

et al. 2001

Analytical Biochemistry

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Section: Discussionmentioning

confidence: 98%

A Fluorimetric Method Based on Changes in Membrane Potential for Screening Paralytic Shellfish Toxins in Mussels

Louzao

Vieytes

Sousa

et al. 2001

Analytical Biochemistry

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“…Effects of SIN-1 on membrane potential of SH-SY5Y cells was monitored with the membrane potential-dependent fluorescent dye Oxonal V, as described by Kukkonen et al (1996). In brief, cells were harvested in PBS containing 0.02% EDTA, and then they were centrifuged and washed once with Ca 2ϩ -free KRH buffer.…”

Section: Methodsmentioning

confidence: 99%

Activity and Subcellular Trafficking of the Sodium-Coupled Choline Transporter CHT Is Regulated Acutely by Peroxynitrite

Pinthong

Black²,

Ribeiro³

et al. 2007

Mol Pharmacol

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Excess formation of nitric oxide and superoxide by-products (peroxynitrite, reactive oxygen, and reactive nitrogen species) attenuates cholinergic transmission potentially having a role in Alzheimer disease pathogenesis. In this study, we investigated mechanisms by which acute exposure to peroxynitrite impairs function of the sodium-dependent hemicholinium-3 (HC-3)-sensitive choline transporter (CHT) that provides substrate for acetylcholine synthesis. The peroxynitrite generator 3-morpholinosydnonimine (SIN-1) acutely inhibited choline uptake in cells stably expressing FLAG-tagged rat CHT in a dose-and timedependent manner, with an IC 50 ϭ 0.9 Ϯ 0.14 mM and t 1/2 ϭ 4 min. SIN-1 significantly reduced V max of choline uptake without altering the K m . This correlated with a SIN-1-induced decrease in cell surface CHT protein, observed as lowered levels of HC-3 binding and biotinylated CHT at the plasma membrane. It is noteworthy that short-term exposure of cells to SIN-1 accelerated the rate of internalization of CHT from the plasma membrane, but it did not alter return of CHT back to the cell surface. SIN-1 did not disrupt cell membrane integrity or cause cell death. Thus, the inhibitory effect of SIN-1 on choline uptake activity and HC-3 binding was related to enhanced internalization of CHT proteins from the plasma membrane to subcellular organelles.

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“…In order to provide a good screening for the effect of gambierol on excitable cells, we designed a couple of fluorimetric approaches based on: i) measurement of changes in membrane potential by using the dye bisoxonol and ii) quantification of intracellular calcium by loading with fura-2 AM. For this matter, neuroblastoma cultures are an excellent model to distinguish the mechanism of action of different neurotoxins [26,27].…”

Section: Discussionmentioning

confidence: 99%

The Sodium Channel of Human Excitable Cells is a Target for Gambierol

Louzao

Cagide

Vieytes

et al. 2006

Cell Physiol Biochem

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Background: Gambierol is a polycyclic ether toxin with the same biogenetic origin as ciguatoxins. Gambierol has been associated with neurological symptoms in humans even though its mechanism of action has not been fully characterized. Methods: We studied the effect of gambierol in human neuroblastoma cells by using bis-oxonol to measure membrane potential and FURA-2 to monitor intracellular calcium. Results: We found that this toxin: i) produced a membrane depolarization, ii) potentiated the effect of veratridine on membrane potential iii) decreased ciguatoxininduced depolarization and iv) increased cytosolic calcium in neuroblastoma cells. Conclusion: These results indicate that gambierol modulate ion fluxes by acting as a partial agonist of sodium channels.

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Muscarinic depolarization of SH-SY5Y human neuroblastoma cells as determined using oxonol V

Cited by 5 publications

References 24 publications

A Fluorimetric Method Based on Changes in Membrane Potential for Screening Paralytic Shellfish Toxins in Mussels

A Fluorimetric Method Based on Changes in Membrane Potential for Screening Paralytic Shellfish Toxins in Mussels

Activity and Subcellular Trafficking of the Sodium-Coupled Choline Transporter CHT Is Regulated Acutely by Peroxynitrite

The Sodium Channel of Human Excitable Cells is a Target for Gambierol

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