Paradoxical modulation of short-term facilitation of dopamine release by dopamine autoreceptors

Electrophysiological studies have demonstrated that dopaminergic neurons burst fire during certain aspects of reward-related behavior; however, the correlation between dopamine release and cell firing is unclear. When complex stimulation patterns that mimic intracranial self-stimulation were employed, dopamine release was shown to exhibit facilitated as well as depressive components (Montague et al. 2004). Understanding the biological mechanisms underlying these variations in dopamine release is necessary to unravel the correlation between unit activity and neurotransmitter release. The dopamine autoreceptor provides negative feedback to dopamine release, inhibiting release on the time scale of a few seconds. Therefore, we investigated this D(2) receptor to see whether it is one of the biological mechanisms responsible for the history-dependent modulation of dopamine release. Striatal dopamine release in anesthetized rats was evoked with stimulus trains that were designed to promote the variability of dopamine release. Consistent with the well established D(2)-mediated autoinhibition, the short-term depressive component of dopamine release was blocked by raclopride, a D(2) antagonist, and enhanced by quinpirole, a D(2)-receptor agonist. Surprisingly, these same drugs exerted a similar effect on the short-term facilitated component: a decrease with raclopride and an increase with quinpirole. These data demonstrate that the commanding control exerted by dopamine autoreceptors over short-term neuroadaptation of dopamine release involves both inhibitory and paradoxically, facilitatory components.     

Relationship between stem CO2 efflux, stem sap velocity and xylem CO2 concentration in young loblolly pine trees

We measured diel patterns of stem surface CO2 efflux (Es, micromol m(-2) s(-1)), sap velocity (vs, mm s(-1)) and xylem CO2 concentration ([CO2]) (Xs, %) in 8-year-old loblolly pine trees during the spring to determine how vs and Xs influence Es. All trees showed a strong diel hysteresis between Es and stem temperature, where at a given temperature, Es was lower during the day than at night. Diel variations in temperature-independent Es were correlated with vs (R2= 0.54), such that at maximum vs, Es was reduced between 18 and 40%. However, this correlation may not represent a cause-and-effect relationship. In a subset of trees, vs was artificially reduced by progressively removing the tree canopy. Reducing vs to near zero had no effect on Es and did not change the diel hysteretic response to temperature. Diel Xs tended to decrease with vs and increase with Es, however, in defoliated trees, large increases in Xs, when vs approximately 0, had no effect on Es. We conclude that at this time of the year, Es is driven primarily by respiration of cambium and phloem tissues and that sap flow and xylem transport of CO2 had no direct influence on Es.     

The particle inflow gun can be used to co-transform Paramecium using Tungsten particles

A particle inflow gun (PIG) was constructed and tested for its utility to transform Paramecium using tungsten or gold as the DNA carrier particle. In the first set of experiments we transformed Paramecium with a plasmid containing the neomycin-resistance gene, obtaining a transformation efficiency of 0.31+/-0.14% (mean+/-SD) for tungsten particles and 1.30+/-0.29% for gold particles. Plasmid DNA precipitated upon tungsten was shown to be stable for transformation purposes for up to 1 h prior to use and had no detectable effects on transformation efficiency. In addition, we demonstrated that at high frequency (71+/-20%) a Paramecium mutant strain could be phenotypically rescued by co-transformation with a second plasmid containing the selectable neomycin-resistance gene. The PIG coupled with tungsten particles as the carrier offers a low-cost alternative for biolistic transformation of Paramecium.     

Gene application with in utero electroporation in mouse embryonic brain

Mouse genetic manipulations, such as the production of gene knock-out, knock-in, and transgenic mice, have provided excellent systems for analysis of numerous genes functioning during development. Nevertheless, the lack of specific promoters and enhancers that control gene expression in specific regions and at specific times, limits usage of these techniques. However, progress in in utero systems of electroporation into mouse embryos has opened a new window, permitting new approaches to answering important questions. Simple injection of plasmid DNA solution and application of electrical current to mouse embryos results in transient area- and time-dependent transfection. Further modification of the technique, arising from variations in types of electrodes used, has made it possible to control the relative size of the region of transfection, which can vary from a few cells to entire tissues. Thus, this technique is a powerful means not only of characterizing gene function in various settings, but also of tracing the migratory routes of cells, due to its high efficiency and the localization of gene expression it yields. We summarize here some of the potential uses and advantages of this technique for developmental neuroscience research.     

Oxygen distribution and movement,respiration and nutrient loading in banana roots (Musa spp. L.) subjected to aerated and oxygen-depleted environments

Excessive soil wetness is a common feature where bananas (Musa spp.) evolved. Under O₂ deficiency, a property of wet soils, root growth and functions will be influenced by the respiratory demand for O₂ in root tissues, the transport of O₂ from the shoot to root and the supply of O₂ from the medium. In laboratory experiments with nodal roots of banana, we examined how these features influenced the longitudinal and radial distributions of O₂ within roots, radial O₂ loss, solute accumulation in the xylem, root hydraulic conductivity, root elongation and root tip survival. In aerated roots, the stele respired about 6 times faster than the cortex on a volume basis. Respiratory O₂ consumption decreased substantially with distance from the root apex and at 300–500 mm it was 80% lower than at the apex. Respiration of lateral roots constituted a sink for O₂ supplied via aerenchyma, and reduced O₂ flow towards the tip of the supporting root. Stelar anoxia could be induced either by lowering the O₂ partial pressure in the bathing medium from 21 to 4 kPa (excised roots) or, in the case of intact roots, by reducing the O₂ concentration around the shoot. The root hair zone sometimes extended to 1.0 mm from the root surface and contributed up to a 60% drop in O₂ concentration from a free-flowing aerated solution to the root surface. There was a steep decline in O₂ concentration across the epidermal-hypodermal cylinder and some evidence of a decline in the O₂ permeability of the epidermal-hypodermal cylinder with increasing distance from the root apex. The differences in O₂ concentration between cortex and stele were smaller than reported for maize and possibly indicated a substantial transfer rate of dissolved O₂ from cortex to stele in banana, mediated by a convective water flow component. An O₂ partial pressure of 4 kPa in the medium reduced net nutrient transfer into the vascular tissue in the stele within 1 or 2 h. Hypoxia also caused a temporary decrease in radial root hydraulic conductivity by an order of magnitude. In O₂ deficient environments, the stele would be among the first tissues to suffer anoxia and O₂ consumption within the root hair zone might be a major contributor to root anoxia/hypoxia in banana growing in temporarily flooded soils.     

Photosynthesis and respiration of a diatom biofilm cultured in a new gradient growth chamber

Abstract A diatom biofilm was grown in a chamber developed for culture of biofilms in chemical gradients. The diatoms grew on a polycarbonate membrane filter which separated a sterile reservoir, with added phosphate, from a reservoir without phosphate. Within 3 weeks of inoculation, a thick biofilm developed on the surface of the filter. The biofilms were homogeneous and therefore suitable for calculations of O₂ diffusion fluxes from concentration profiles of O₂. Profiles of O₂, pH, and gross photosynthesis at different light intensities and liquid medium concentrations of dissolved inorganic carbon and O₂ were measured with microelectrodes. Respiratory activity in a layer of the biofilm was determined as the difference between gross photosynthesis and outflux of O₂ from that layer. The photosynthetic activity in a well-developed biofilm grown at 360 μEinst m⁻² s⁻¹ and 2.4 mM HCO₃⁻ was limited by the supply of inorganic carbon. Exposure to light above 360 μEinst m⁻² s⁻¹ stimulated gross photosynthesis as well as respiratory processes without affecting net outflux of O₂. Higher concentrations of inorganic carbon, on the other hand, enhanced gross photosynthesis without concurrent increase in respiratory rate, resulting in an increased outflux of O₂. High concentrations of O₂ in the liquid medium decreased the net outflux of O₂ with little effect on the gross photosynthesis. The effects of inorganic carbon and O₂ on the metabolic activities of the biofilm were consistent with the presence of photorespiratory activity.     

应用线性硅电极阵列检测海马场电位和单细胞动作电位

近年来,硅材料微电极阵列发展迅速,μ为研究大脑神经细胞活动的时空特性提供了理想的手段.考察了线性硅材料微电极阵列在神经细胞电位检测中的稳定性,以及对于单细胞动作电位检测的有效性.实验结果表明,在麻醉大鼠海马CA1区场电位记录中,上下移动记录微电极200μm,对于正向和反向诱发电位的记录几乎没有影响,说明,线性微电极阵列对于神经细胞的损伤很小,检测性能稳定.电极阵列上处于细胞胞体层的测量点可以有效地记录到CA1神经细胞的动作电位发放,同一记录点上可以清楚地分辨出数个不同神经细胞的发放电位.实验结果显示了硅电极阵列操作简便、检测信号稳定和获取信息多等特点,对于开展微电极阵列应用研究的工作人员具有借鉴作用.     

Simultaneous measurement of ammonium,nitrate and proton fluxes along the length of eucalypt roots

Knowledge of the preferred source of N for Eucalyptus nitens will lead to improved fertiliser management practices in plantations. Ion selective microelectrodes were used non-invasively to measure simultaneously net fluxes of NH₄ ⁺, NO₃ ^– and H⁺ along the tap root of solution-cultured E. nitens. Measurements were conducted in solutions containing 100 m NH₄NO₃. The pattern of fluxes was such that there was a large influx of NH₄ ⁺, a smaller influx of NO₃ ^– and large H⁺ efflux. The ratio of these fluxes was constant, according to the ratio 3:1:–6 (NH₄ ⁺:NO₃ ^–:H⁺). Within the region 20–60 mm from the root apex of E. nitens seedlings there was spatial and temporal variation in fluxes but flux patterns remained constant. Root hair density did not affect fluxes nor did proximity to lateral roots. Variation was less than that found in previous studies of localised root fluxes using similar high-resolution measurement techniques. It was concluded that small-scale spatial variation in fluxes may have confounded previous studies. There were associations between fluxes of all three ions, the strongest associations being between NH₄ ⁺ and H⁺, and NH₄ ⁺ and NO₃ ^–. Overall, these results are consistent with NH₄ ⁺ being the preferred source N for E. nitens.     

Microelectrode ion and O2 fluxes measurements reveal differential sensitivity of barley root tissues to hypoxia

Hypoxia-induced changes in net H+, K+ and O2 fluxes across the plasma membrane (PM) of epidermal root cells were measured using the non-invasive microelectrode ion flux measurement (MIFE) system in elongation, meristem and mature root zones of two barley (Hordeum vulgare L.) varieties contrasting in their waterlogging (WL) tolerance. The ultimate goal of this study was to shed light on the mechanisms underlying effects of WL on plant nutrient acquisition and mechanisms of WL tolerance in barley. Our measurements revealed that functionally different barley root zones have rather different O2 requirements, with the highest O2 influx being in the elongation zone of the root at about 1 mm from the tip. Oxygen deprivation has qualitatively different effects on the activity of PM ion transporters in mature and elongation zones. In the mature zone, hypoxic treatment caused a very sharp decline in K+ uptake in the WL sensitive variety Naso Nijo, but did not reduce K+ influx in the WL tolerant TX9425 variety. In the elongation zone, onset of hypoxia enhanced K+ uptake from roots of both cultivars. Pharmacological experiments suggested that hypoxia-induced K+ flux responses are likely to be mediated by both K(+) -inward- (KIR) and non-selective cation channels (NSCC) in the elongation zone, while in the mature zone K(+) -outward- (KOR) channels are the key contributors. Overall, our results suggest that oxygen deprivation has an immediate and substantial effect on root ion flux patterns, and that this effect is different in WL-sensitive and WL-tolerant cultivars. To what extent this difference in ion flux response to hypoxia is a factor conferring WL tolerance in barley remains to be answered in future studies.     

A METHOD TO IMPROVE THE SPATIAL RESOLUTION OF PHOTOSYNTHETIC RATES OBTAINED BY OXYGEN MICROSENSORS

To determine gross photosynthesis in benthic microalgal communities, oxygen microelectrodes were used to measure the rate of decrease within the first 4 s after extinction of light. Photosynthetic rates calculated from third-order polynomial fits to the curve of decreasing O₂ concentration were compared to the rates obtained by the traditional method, where rates were estimated from linear regression. When photosynthesis was calculated for the fitted initial rates of O₂ decrease, maximum rates in microbial mats were up to 32% higher, and the depth-integrated gross photosynthesis was 5%–10% higher than the rates determined by the traditional method. The determinations from fitted initial rates also resulted in a more detailed profile of photosynthetic rate than that normally obtained. Computer simulation based on diffusion models, where the estimated initial rates of O₂ decrease were assumed to represent actual photosynthesis rates, verified the validity of the curve-fitting procedure for obtaining high-resolution photosynthesis profiles.