Influence of pipecolic acid on the release and uptake of [3H]GABA from brain slices of mouse cerebral cortex

Recently, pipecolic acid (PA) has been involved in the functioning of the GABAergic system. In the present work we have studied the effect of PA on GABA uptake and release in cerebral cortex slices. PA (100 M) was able to increase the release of [³H]GABA (90%) stimulated by mild depolarization with 15 mM potassium. If during the labeling of the tissue with [³H]GABA, -alanine was present, PA also enhanced the release (42%). However, when nipecotic acid was present instead -alanine, no stimulation of [³H]GABA release by potassium was observed neither in the control nor in the presence of PA. Spontaneous release was not affected by PA in any of the experimental conditions tested. In uptake experiments, only when -alanine was present in the medium PA significantly diminished the uptake (36%) of [³H]GABA. These results suggest that the effect of PA is mostly at the presynaptic level, inhibiting the neuronal GABA uptake and/or enhancing its release.     

[3H]Tyramine binding: A comparison with neuronal [3H]-dopamine uptake and [3H]mazindol binding processes

[³H]Spiperone ([³H]SPI) binding sites in rat or bovine striata have been solubilized using CHAPS or digitonin detergents. Solubilized sites retained the binding characteristics of those in native membrane preparations. The same solubilized material, however, did not bind [³H]tyramine ([³H]PTA), thus indicating that [³H]PTA binding sites and DA receptors are different chemico-physical entities. In membrane preparations or crude synaptosomes obtained from the c.striatum of neonatally-rendered hypothyroid rats, when central DA-pathways are impaired, both [³H]PTA binding and [³H]DA uptake processes were markedly decreased, with no effect on [³H]mazindol ([³H]MAZ) binding, compared to euthyroids. Reserpine, a well-known inhibitor of DA-uptake into a variety of secretory vesicles, and a potent in vivo andin vitro inhibitor of [³H]PTA binding, did not affect the [³H]MAZ binding process. This further supported the suggestion that while [³H]PTA binding sites are almost totally associated with the vesicular transporter for DA, [³H]MAZ does label a site involved in the DA-translocation across the neuronal membrane. The latter process seems to be rather insensitive to thyroid hypofunction, when however the intracellular storage of DA might be consistently impaired. In conclusion, PTA might be well exploited as a marker of the DA vesicular transporter through its molecular characterization, whenever possible.Special issue dedicated to Dr. Paola S. Timiras     

β-Dl-Methylene-aspartate,an inhibitor of aspartate aminotransferase,potently inhibitsl-glutamate uptake into astrocytes

[³H]Glutamate uptake into astrocytes in primary culture was potently inhibited by the aspartate analoguesl- andd-aspartic acid,Dl-threo--hydroxy-aspartic acid,l-aspartic acid--hydroxymate (IC50's: 136, 259, 168, and 560 M, respectively) and by -Dl-methylene-aspartate, a suicide inhibitor of asparate aminotransferase (IC50: 524 M), and by the endogenous sulphur-containing amino acidl-cysteinesulfinic acid (IC50: 114 M). [³H]Glutamate uptake was not significantly affected by either N-methyl-d-aspartate orDl-homocysteine thiolactone. These results demonstrate that other excitatory amino acids including aspartate andl-cysteinesulfinic acid (but excludingl-homocysteic acid) interact with the glutamate transport system of astrocytes. Inhibition of glutamate uptake may significantly increase the level of neuronal excitability.     

Influence of temperature on root development and phosphate influx in winter wheat grown at different P levels

Root development was studied in winter wheat ( Triticum aestivum L. cv Starke II) grown at 5,10, 15 and 20°C in nutrient solutions with phosphate concentrations of 10, 100 or 1000 μM . The plants were grown for 38 days (5 and 10°C), 19 days (15°C) or 14 days (20°C). At the end of the cultivation period the phosphate influx in the roots was determined with ³²P-phosphate. Root development (lateral and seminal roof length and number) was monitored throughout the cultivation period on the same individuals by repeated (approximately every second day) photocopying of the roots for measurements with digitizer and appropriate software. The 5°C treatment yielded no laterals, and the seminals were only slightly affected by the different phosphate treatments. The 10 μM phosphate treatment gave high root:shoot dry weight ratio, high average lateral root length and high specific root length [m root (g root fresh weight)^-1]. The 1000 μM phosphate treatment yielded the highest number of laterals per m seminal root, and usually also the highest absolute numbers. Phosphate influx decreased with increased P status of the roots. It is argued that phosphate influx is dependent on factors such as P status, root geometry and relative root extension rate.     

Agrobacterium-mediated gene delivery and the biology of hostrange limitations

Insertion of foreign DNA into Ti plasmid-derived vectors in Agrobacterium tumefaciens is currently the most frequently used strategy for generating transgenic plants in a wide variety of species. Limitations of the host range of Agrobacterium restrict its usefulness in many cases, particularly when dealing with monocotyledonous plants. The objective of this presentation is to briefly discuss the efficiency of the transformation process utilized by Agrobacterium tumefaciens , potential barriers to efficient transformation by Agrobacterium that result in limitation of its useful host range, and how an understanding of the successful Agrobacterium /plant cell interaction might lead to advances in a variety of DNA delivery methodologies.     

Frankia strains of soil underBetula pendula: Behaviour in soil and in pure culture

A K/Rb isotope dilution method was used to determine the uptake of K from undisturbed subsoils. Rb was applied to the topsoil (0–30 cm) to trace the K taken up from the topsoil by crops. The K/Rb ratio in the crops increases when roots contact the Rb-free subsoil. This change in the K/Rb ratio enables the calculation of the uptake of K from the subsoil. Results of 34 field experiments on loess-parabrown soils in N. Germany showed that the subsoil (>30 cm) supplied, on average, 34% of the total K uptake by spring wheat (range 9–70%). The range between the experimental sites is considered in relation to the contents of K in the top and subsoils (as extracted by 0.025 N CaCl₂ solution), the proportion of the total root length in the subsoils, and competition for K between roots in the top and subsoil. In subsoils with similar K contents, uptake from the subsoil decreased significantly from 65 to 21% of total K uptake, as K contents in the topsoils increased from 4 to 8 mg K/100 g. On sites with the same K contents in topsoils (9 mg K/100 g), the subsoil supplied 12 to 61% of total K uptake as the K contents of the subsoil increased from 2 to 27 mg K/100 g. The contribution of uptake of K from the subsoil increased with the development of the crop, from 8% at first node stage to 35% at ear emergence, as the proportion of total root length in the subsoil increased. High root length densities in the topsoil (9 cm/cm³) resulted in competition for K between roots and increased uptake of K from the subsoil.     

Net Mg2+ uptake in relation to the amount of exchangeable Mg2+ in the Donnan free space of ryegrass roots

Ammonium acetate and BaCl₂-triethanolamine were used to desorb Mg²⁺ from the root Donnan free space (DFS) of 23-d-old ryegrass (Lolium multiflorum Lam. cvs. Gulf and Wilo). Amounts of desorbed Mg²⁺ increased with the increase in Mg²⁺ activity of the nutrient solution. Slightly less Mg²⁺ was desorbed by Ba²⁺ than by NH₄ ⁺. Previously published data on short-term net Mg²⁺ uptake by intact 23-d-old ryegrass plants of the two cultivars were linearly related to the amount of exchangeable Mg⁺ desorbed from the root DFS (r²=0.90 and 0.81 for the desorption by NH₄ ⁺ and Ba²⁺, respectively). A sward of Mg²⁺ ions attracted to the negative charges of the cell surface is suggested to represent a part of a pool of Mg²⁺ available for active transport through the plasmalemma.     

Nitrification and nitrate uptake: Leaching balance in a declined forest ecosystem in eastern France

Nitrate uptake and leaching were measured during one year in a declined fir forest on the Vosges highlands (eastern France), in order to investigate whether excess nitrification could be responsible for a deleterious acidification of the ecosystem. Nitrate uptake by the vegetation was active mainly from spring to early fall, and then reached about 66 kg N ha^-1. No significant leaching loss occurred during the growth period of the vegetation. Significant nitrate leaching occurred in winter (about 17 kg N ha^-1). During fall and winter the nitrification rate was of the same magnitude as values reported for other ecosystems, and, thus, was not considered to be abnormaly strong. No abnormal temporal discoupling of nitrate production and nitrate uptake occurred in the ecosystem, and forest decline must therefore have some other cause.     

Uptake of phosphate from soils and fertilizers as affected by soil P availability and solubility of phosphorus fertilizers

Fertilizers labelled with ³²P were used to measure amounts of phosphorus, P_s and P_F, taken up by Lolium perenne from available soil P and from P fertilizer respectively, when applied at a rate of 66 mg P·(kg soil^–1) in greenhouse experiments. The quantity P_s of phosphorus taken up from soil in the presence of P fertilizer was compared to the quantity P_o taken up from soil without P fertilizer. The quantity (P_s–P_o) is positive for low P_o values, i.e. in soils poor in available phosphorus, but is negative for high P_o values indicating that an input of P fertilizer can induce a decrease in the utilization of available soil phosphorus. Moreover, for a given soil, the quantity (P_s–P_o) depends on the chemical form of the fertilizer. The standard method of evaluation of P fertilizer efficiency is based on the assumption that P_s=P_o, but P_s can differ from P_o. This result can explain the contradictory data published from field experiments about the efficiency of the various P fertilizers.     

Zn buffering capacity and Zn accumulation in Swiss chard for some sludge-amended soils

Zinc sorption by soils can greatly affect its availability to plants. This study was conducted to determine the relationship between the Zn sorption capacity and plant Zn accumulation in five sludge-amended soils using Swiss chard (Beta vulgaris L.) as an indicator plant. Zinc sorption as a function of Zn concentration and pH was determined for the soils which received no sludge amendment; also DTPA (diethylenetriaminepentaacetic acid) extractable Zn was determined in all soils. Whereas the responses of DTPA-Zn and plant Zn to pH and the quantities of Zn sorbed were similar, the logarithm of DTPA-Zn accounted for only 82% of the variability in the logarithm of Zn accumulation by the plants. The variability was better explained when pH was included with DTPA-Zn in stepwise multiple regressions. The Zn buffering capacity, defined as the ratio of the change in quantity of Zn sorbed ( Zn_s) to the change in Zn solution concentration (Zn₁) (or Zn_s/Zn₁), and the estimated quantity of Zn sorbed were used as a basis to measure Zn intensity. Zinc intensity, which reflects Zn solution concentration, was the predominant factor controlling Zn accumulation by Swiss chard, judging from the good fit of the values of both parameters to the Michaelis-Menten equation. The maximum Zn accumulation was approximately 9 mmol kg^–1.Scientific paper no. 8901-29, Department of Agronomy and Soils, College of Agriculture and Home Economics Research Center. Washington State University, Pullman, WA 99164, USA.Scientific paper no. 8901-29, Department of Agronomy and Soils, College of Agriculture and Home Economics Research Center. Washington State University, Pullman, WA 99164, USA.