- Title
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General anesthetics protects against cardiac arrest-induced brain injury by inhibiting calcium wave propagation in zebrafish
- Authors
- Xu, D.J., Wang, B., Zhao, X., Zheng, Y., Du, J.L., Wang, Y.W.
- Source
- Full text @ Mol. Brain
Cardiac arrest causes a burst of calcium wave in brain. a Schematic showing the experimental scheme. b Schematic showing cardiac arrest (CA) model in zebrafish. A glass micropipette was used to press zebrafish heart (red) until the heart stop beating, neuronal activity was examined with in vivo time-lapse confocal imaging after CA. c Calcium activity of Tg (HuC:GCaMP5) transgenic zebrafish in different brain regions after CA (lateral view). ROI1 (red ellipses), ROI2 (green ellipses) and ROI3 (yellow ellipses) marked neuron in hindbrain, midbrain and telencephalon, respectively. Scale bar: 100 μm. d Calcium activity of three regions of interest in (c) at a higher time resolution. The arrows mark the onset of calcium activities in each region. e Time-lapse confocal images showing the lateral view of the brain in live Tg (HuC:GCaMP5) transgenic zebrafish in control and CA zebrafish at 7 day post-fertilization (dpf). 0 s indicating calcium wave onset. Scale bar: 100 μm. f The change in fluorescence intensity of neuron in midbrain in individual zebrafish during CA. g Occurrence probability of calcium waves in control and CA zebrafish (*** P<0.001, chi-square test). h Quantification of the latency of CA to calcium wave generation in CA zebrafish. Numbers in the histograms represent the number of embryos analyzed in each group PHENOTYPE:
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Midazolam or ketamine pretreatment inhibits cardiac arrest-induced calcium wave propagation. a-d Time-lapse confocal images showing the lateral view of the brain in live Tg (HuC:GCaMP5) transgenic zebrafish in control (a), CA (b), CA + midazolam (0.5 mM)(c) and CA + ketamine (2.5 mM) pretreatment (d) zebrafish at 7 dpf. 0 s indicating calcium wave onset. MDZ, midazolam. Scale bar: 100 μm. e Occurrence probability of calcium waves in each group (*** P<0.001, chi-square test). Numbers in the histograms represent the number of embryos analyzed in each group PHENOTYPE:
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Midazolam or ketamine pretreatment decreases the neuronal apoptosis after cardiac arrest. a-d TUNEL staining of control (a), CA (b), CA + midazolam (0.5 mM)(c) and CA + ketamine (2.5 mM) pretreatment (d) zebrafish at 7 dpf. Red, TUNEL staining, indicating the apoptotic cell; Blue, DAPI-stained nuclei. Scale bar: 10 μm. e Quantification of percentages of TUNEL positive cells in each group (mean ± s.e.m.; *** P<0.001, one-way ANOVA plus Newman–Keuls multiple comparison test). Numbers in the histograms represent the number of sections analyzed in each group PHENOTYPE:
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