Xenon inhibits excitatory but not inhibitory transmission in rat spinal cord dorsal horn neurons

<p>Abstract</p> <p>Background</p> <p>The molecular targets for the promising gaseous anaesthetic xenon are still under investigation. Most studies identify <it>N</it>-methyl-D-aspartate (NMDA) receptors as the primary molecular target for xenon, but the role of α-amino-3-hydroxy-5-methyl-4-isoxazole-4-propionic acid (AMPA) receptors is less clear. In this study we evaluated the effect of xenon on excitatory and inhibitory synaptic transmission in the superficial dorsal horn of the spinal cord using <it>in vitro </it>patch-clamp recordings from rat spinal cord slices. We further evaluated the effects of xenon on innocuous and noxious stimuli using <it>in vivo </it>patch-clamp method.</p> <p>Results</p> <p><it>In vitro</it>, xenon decreased the amplitude and area under the curve of currents induced by exogenous NMDA and AMPA and inhibited dorsal root stimulation-evoked excitatory postsynaptic currents. Xenon decreased the amplitude, but not the frequency, of miniature excitatory postsynaptic currents. There was no discernible effect on miniature or evoked inhibitory postsynaptic currents or on the current induced by inhibitory neurotransmitters. <it>In vivo</it>, xenon inhibited responses to tactile and painful stimuli even in the presence of NMDA receptor antagonist.</p> <p>Conclusions</p> <p>Xenon inhibits glutamatergic excitatory transmission in the superficial dorsal horn <it>via </it>a postsynaptic mechanism. There is no substantial effect on inhibitory synaptic transmission at the concentration we used. The blunting of excitation in the dorsal horn lamina II neurons could underlie the analgesic effect of xenon.</p

Selective activation of primary afferent fibers evaluated by sine-wave electrical stimulation

Transcutaneous sine-wave stimuli at frequencies of 2000, 250 and 5 Hz (Neurometer) are thought to selectively activate Aβ, Aδ and C afferent fibers, respectively. However, there are few reports to test the selectivity of these stimuli at the cellular level. In the present study, we analyzed action potentials (APs) generated by sine-wave stimuli applied to the dorsal root in acutely isolated rat dorsal root ganglion (DRG) preparations using intracellular recordings. We also measured excitatory synaptic responses evoked by transcutaneous stimuli in substantia gelatinosa (SG) neurons of the spinal dorsal horn, which receive inputs predominantly from C and Aδ fibers, using in vivo patch-clamp recordings. In behavioral studies, escape or vocalization behavior of rats was observed with both 250 and 5 Hz stimuli at intensity of ~0.8 mA (T5/ T250), whereas with 2000 Hz stimulation, much higher intensity (2.14 mA, T2000) was required. In DRG neurons, APs were generated at T5/T250 by 2000 Hz stimulation in Aβ, by 250 Hz stimulation both in Aβ and Aδ, and by 5 Hz stimulation in all three classes of DRG neurons. However, the AP frequencies elicited in Aβ and Aδ by 5 Hz stimulation were much less than those reported previously in physiological condition. With in vivo experiments large amplitude of EPSCs in SG neurons were elicited by 250 and 5 Hz stimuli at T5/ T250. These results suggest that 2000 Hz stimulation excites selectively Aβ fibers and 5 Hz stimulation activates noxious transmission mediated mainly through C fibers. Although 250 Hz stimulation activates both Aδ and Aβ fibers, tactile sensation would not be perceived when painful sensation is produced at the same time. Therefore, 250 Hz was effective stimulus frequency for activation of Aδ fibers initiating noxious sensation. Thus, the transcutaneous sine-wave stimulation can be applied to evaluate functional changes of sensory transmission by comparing thresholds with the three stimulus frequencies

Impacts of regional mixing on the temperature structure of the equatorial Pacific Ocean. Part 1: Vertically uniform vertical diffusion

AbstractWe investigate the sensitivity of numerical-model solutions to regional changes in vertical diffusion. Specifically, we vary the background diffusion coefficient, κb, within spatially distinct subregions of the tropical Pacific, assess the impacts of those changes, and diagnose the processes that account for them.Solutions respond to a diffusion anomaly, δκb, in three ways. Initially, there is a fast response (several months), due to the interaction of rapidly-propagating, barotropic and gravity waves with eddies and other mesoscale features. It is followed by a local response (roughly one year), the initial growth and spatial pattern of which can be explained by one-dimensional (vertical) diffusion. At this stage, temperature and salinity anomalies are generated that are either associated with a change in density (“dynamical” anomalies) or without one (“spiciness” anomalies). In a final adjustment stage, the dynamical and spiciness anomalies spread to remote regions by radiation of Rossby and Kelvin waves and by advection, respectively.In near-equilibrium solutions, dynamical anomalies are generally much larger in the latitude band of the forcing, but the impact of off-equatorial forcing by δκb on the equatorial temperature structure is still significant. Spiciness anomalies spread equatorward within the pycnocline, where they are carried to the equator as part of the subsurface branch of the Pacific Subtropical Cells, and spiciness also extends to the equator via western-boundary currents. Forcing near and at the equator generates strong dynamical anomalies, and sometimes additional spiciness anomalies, at pycnocline depths. The total response of the equatorial temperature structure to δκb in various regions depends on the strength and spatial pattern of the generation of each signal within the forcing region as well as on the processes of its spreading to the equator

Efficient and Directive Generation of Two Distinct Endoderm Lineages from Human ESCs and iPSCs by Differentiation Stage-Specific SOX17 Transduction

The establishment of methods for directive differentiation from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) is important for regenerative medicine. Although Sry-related HMG box 17 (SOX17) overexpression in ESCs leads to differentiation of either extraembryonic or definitive endoderm cells, respectively, the mechanism of these distinct results remains unknown. Therefore, we utilized a transient adenovirus vector-mediated overexpression system to mimic the SOX17 expression pattern of embryogenesis. The number of alpha-fetoprotein-positive extraembryonic endoderm (ExEn) cells was increased by transient SOX17 transduction in human ESC- and iPSC-derived primitive endoderm cells. In contrast, the number of hematopoietically expressed homeobox (HEX)-positive definitive endoderm (DE) cells, which correspond to the anterior DE in vivo, was increased by transient adenovirus vector-mediated SOX17 expression in human ESC- and iPSC-derived mesendoderm cells. Moreover, hepatocyte-like cells were efficiently generated by sequential transduction of SOX17 and HEX. Our findings show that a stage-specific transduction of SOX17 in the primitive endoderm or mesendoderm promotes directive ExEn or DE differentiation by SOX17 transduction, respectively

Toward Regional Characterizations of the Oceanic Internal Wavefield

Many major oceanographic internal wave observational programs of the last 4 decades are reanalyzed in order to characterize variability of the deep ocean internal wavefield. The observations are discussed in the context of the universal spectral model proposed by Garrett and Munk. The Garrett and Munk model is a good description of wintertime conditions at Site-D on the continental rise north of the Gulf Stream. Elsewhere and at other times, significant deviations in terms of amplitude, separability of the 2-D vertical wavenumber - frequency spectrum, and departure from the model's functional form are noted. Subtle geographic patterns are apparent in deviations from the high frequency and high vertical wavenumber power laws of the Garrett and Munk spectrum. Moreover, such deviations tend to co-vary: whiter frequency spectra are partnered with redder vertical wavenumber spectra. Attempts are made to interpret the variability in terms of the interplay between generation, propagation and nonlinearity using a statistical radiative balance equation. This process frames major questions for future research with the insight that such integrative studies could constrain both observationally and theoretically based interpretations

Yusho patients show increased serum IL-17, IL-23, IL-1β, and TNFα levels more than 40 years after accidental polychlorinated biphenyl poisoning

The Yusho poisoning incident, caused by rice oil contaminated with polychlorinated biphenyls (PCBs), polychlorinated quarterphenyls (PCQs), and polychlorinated dibenzofurans (PCDFs) generated by heat-denatured PCBs, occurred in 1968 in western Japan. Although severe symptoms are rarely observed today, the levels of PCBs and PCDFs in the sera of Yusho patients remain high. The aryl hydrocarbon receptor (AhR), which also acts as a dioxin receptor, is a transcriptional regulator that mediates dioxin toxicity. Recent studies show that dioxin mediates its immune toxic effects via AhR and that AhR activation induces dysregulation of interleukin (IL)-17-producing T (TH17) cells. This study therefore hypothesized that Yusho patients would show dysregulated TH17 cell-mediated immune responses. To validate the hypothesis, levels of IL-17 and IL-22, each secreted by TH17 cells, along with IL-1β and IL-23 were measured in serum samples from 40 Yusho patients and 40 age-matched controls. Levels of tumor necrosis factor (TNF)-α potentially secreted by TH17 cell-stimulated neutrophils and macrophages were also measured. The results indicated that serum IL-17 levels, as well as those of IL-1β, IL-23, and TNFα, were significantly higher in Yusho patients than in controls. In contrast, serum IL-22 levels were significantly lower in the Yusho patients. These results suggest that Yusho patients have dysregulated TH17 cell-mediated immune responses that may be linked to inflammation

BMP4 induction of trophoblast from mouse embryonic stem cells in defined culture conditions on laminin

Because mouse embryonic stem cells (mESCs) do not contribute to the formation of extraembryonic placenta when they are injected into blastocysts, it is believed that mESCs do not differentiate into trophoblast whereas human embryonic stem cells (hESCs) can express trophoblast markers when exposed to bone morphogenetic protein 4 (BMP4) in vitro. To test whether mESCs have the potential to differentiate into trophoblast, we assessed the effect of BMP4 on mESCs in a defined monolayer culture condition. The expression of trophoblast-specific transcription factors such as Cdx2, Dlx3, Esx1, Gata3, Hand1, Mash2, and Plx1 was specifically upregulated in the BMP4-treated differentiated cells, and these cells expressed trophoblast markers. These results suggest that BMP4 treatment in defined culture conditions enabled mESCs to differentiate into trophoblast. This differentiation was inhibited by serum or leukemia inhibitory factor, which are generally used for mESC culture. In addition, we studied the mechanism underlying BMP4-directed mESC differentiation into trophoblast. Our results showed that BMP4 activates the Smad pathway in mESCs inducing Cdx2 expression, which plays a crucial role in trophoblast differentiation, through the binding of Smad protein to the Cdx2 genomic enhancer sequence. Our findings imply that there is a common molecular mechanism underlying hESC and mESC differentiation into trophoblast