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Propofol alters vesicular transport in rat cortical neuronal cultures

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Turina D., Bjornstrom K., Sundovist T., Eintrei C.
Journal of Physiology and Pharmacology
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2011
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tom 62
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nr 1
Neuronal intracellular transport is performed by motor proteins, which deliver vesicles, organelles and proteins along cytoskeletal tracks inside the neuron. We have previously shown that the anesthetic propofol causes dose- and time-dependent, reversible retraction of neuronal neurites. We hypothesize that propofol alters the vesicular transport of cortical neurons due to this neurite retraction. Primary cultures of co-cultivated rat cortical neurons and glial cells were exposed to either 2 µM propofol, control medium or the lipid vehicle, in time-response experiments. Reversibility was tested by washing propofol off the cells. The role of the GABAA receptor (GABAAR) was assessed with the GABAAR antagonist gabazine. Vesicles were tracked using differential interference contrast video microscopy. Propofol caused a retrograde movement in 83.4±5.2% (mean±S.E.M.) of vesicles, which accelerated over the observed time course (0.025±0.012 µm·s-1). In control medium, vesicles moved predominantly anterograde (84.6±11.1%) with lower velocity (0.011±0.004 µm·s-1). Cells exposed to the lipid vehicle showed the same dynamic characteristics as cells in control medium. The propofol-induced effect on vesicle transport was reversible and blocked by the GABAAR antagonist gabazine in low concentration. Our results show that propofol causes a reversible, accelerating vesicle movement toward the neuronal cell body that is mediated via synaptic GABAAR. We have previously reported that propofol initiates neurite retraction, and we propose that propofol causes vesicle movement by retrograde flow of cytoplasm from the narrowed neurite.
2
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Orexin A reverses propofol and thiopental induced cytoskeletal rearrangement in rat neurons

100%
Turina D., Gerhardsson H., Bjornstrom K.
Journal of Physiology and Pharmacology
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2014
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tom 65
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nr 4
3
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Characterisation of the signal transduction cascade caused by propofol in rat neurons: from the GABAA receptor to thy cytoskeleton

76%
Bjornstrom K., Turina D., Loverock A., Lundgren S., Wijkman M., Lindroth M., Eintrei C.
Journal of Physiology and Pharmacology
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2008
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tom 59
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nr 3
617-632
The anaesthetic propofol interacts with the GABAA receptor, but its cellular signalling pathways are not fully understood. Propofol causes reorganisation of the actin cytoskeleton into ring structures in neurons. Is this reorganisation a specific effect of propofol as apposed to GABA, and which cellular pathways are involved? We used fluorescence-marked actin in cultured rat neurons to evaluate the percentage of actin rings caused by propofol or GABA in combination with rho, rho kinase (ROK), PI3-kinase or tyrosine kinase inhibitors, with or without the presence of extracellular calcium. Confocal microscopy was performed on propofol-stimulated cells and changes in actin between cellular compartments were studied with Western blot. Propofol (3 µg·ml-1), but not GABA (5 µM), caused transcellular actin ring formation, that was dependent on influx of extracellular calcium and blocked by rho, ROK, PI3-kinase or tyrosine kinase inhibitors. Propofol uses rho/ROK to translocate actin from the cytoskeleton to the membrane and its actin ring formation is dependent on an interaction site close to the GABA site on the GABAA receptor. GABA does not cause actin rings, implying that this is a specific effect of propofol.
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