Biochemical and Temporal Analysis of Events Associated with Apoptosis Induced by Lowering the Extracellular Potassium Concentration in Mouse Cerebellar Granule Neurons

Abstract: We analyzed biochemically and temporally the molecular events that occur in the programmed cell death of mouse cerebellar granule neurons deprived of high potassium levels. An hour after switching the neurons to a low extracellular K⁺ concentration ([K⁺]_o), a significant part of the genomic DNA was already cleaved to high-molecular-weight fragments. This phenomenon was intensified with the progression of the death process. Addition of cycloheximide to the neurons 4 h after high [K⁺]_o deprivation resulted in no cell loss and complete recovery of the damaged DNA. DNA margination and nuclear fragmentation as assessed by 4,6-diaminodiphenyl-2-phenylindole staining were observable in a few cells beginning ～4 h after the removal of high [K⁺]_o and developed to nuclear condensation 4 h later. Six hours after high [K⁺]_o deprivation, the DNA was fragmented into oligonucleosome-sized fragments. Within 6 h after removal of the extracellular K⁺, 50% of the neurons were committed to die and lost their ability to be rescued by readministration of 25 mM [K⁺]_o. Similar to high [K⁺]_o deprivation, inhibition of RNA or protein synthesis failed to halt neuronal degeneration of a similar percentage of cells 6 h after the onset of the death process. Mitochondrial function steadily decreased after [K⁺]_o removal. An ～40% decrease in RNA and protein synthesis was detected by 6 h of [K⁺]_o removal during the period of cell death commitment; rates continued to decline gradually thereafter. The temporal characteristics of the DNA damage and recovery, DNA cleavage to oligonucleosome-sized fragments, and the reduction in mitochondrial activity—events that occurred within the critical time—may indicate that these processes have an important part in the mechanism that committed the neurons to die.     

Effects of vesicular-arbuscular mycorrhiza on Tagetes erecta and Zinnia elegans

 Seeds of Tagetes erecta and Zinnia elegans were planted in soil inoculated with the vesicular-arbuscular mycorrhizal fungus Glomus etunicatum. This procedure produced positive effects on both Tagetes and Zinnia compared with the control: faster flowering and an increased number of flowers. Shoot height, shoot and root dry weights and percentage infected root length were also measured. The reactions of these plants to mycorrhizal infection were shown to be independent of changes in the phosphorus, potassium and sodium contents of the plants. Accepted: 1 August 1995     

Relationships of bio-climatic zones and lithology with various characteristics of forest soils in Greece

Summary Soils derived from a number of different parent materials (lithologies) and developed along a climatic gradient, manifested by the altitudinal succession of natural vegetation zones (Mediterranean, sub-Mediterranean, Mountainous and Pseudoalpine), were sampled throughout mainland Greece.In soils derived from siliceous parent materials low in clay, acidity increase and percent base saturation decreases from the Mediterranean to the Pseudoalpine vegetation zones. Clay illuviation is found mainly in soils developed in the Mediterranean and the sub-Mediterranean zones. No such changes are apparent in clayey soils rich in bases.Organic matter content of the mineral portion of the soil profile increases by a factor of 2 with a decrease in mean annual air temperature of about 10°C. The pattern of change in clay and soil organic matter content with climate is in relatively good agreement with soil development trends in the area, when soil profiles are named according to the FAO-Unesco soil map of the world.Concentrations of Ca and Mg decrease and those of total N, total and extractable P, K, Fe, Mn and Zn increase from the Mediterranean to the Mountainous zone. Within the same zone, however, concentrations of N, Ca, K, Fe, Mn and Zn decrease, but those of Mg, total and extractable P increase with soil depth. The concentrations of most macro- and micronutrients in the humic horizon are several times higher than those in the mineral portion of the soil profile due to biological enrichment.     

Potassium accumulation in growing Methanobacterium thermoautotrophicum and its relation to the electrochemical proton gradient

Cultures of Methanobacterium thermoautotrophicum (Marburg) growing on media low in potassium accumulated the cation up to a maximal concentration gradient ([K⁺]_{intracellular}/[K⁺]_{extracellular}) of approximately 50,000-fold. Under these conditions, the membrane potential was determined by measuring the equilibrium distribution of the lipophilic cation (¹⁴C) tetraphenylphosphonium (TPP⁺). This cation was accumulated by the cells 350-to 1,000-fold corresponding to a membrane potential (inside negative) of 170–200 mV. The pH gradient, as measured by equilibrium distribution of the weak acid, benzoic acid, was found to be lower than 0.1 pH units (extracellular pH=6.8). The addition of valinomycin (0.5–1 nmol/mg cells) to the culture reduced the maximal concentration gradient of potassium from 50,000-to approximately 500-fold, without changing the membrane potential. After dissipation of the membrane potential by the addition of ¹²C-TTP⁺ (2 mol/mg cells) or tetrachlorosalicylanilide (3 nmol/mg cells), a rapid and complete efflux of potassium was observed.These data indicate that potassium accumulation in the absence of valinomycin is not in equilibrium with the membrane potential. It is concluded that at low extracellular K⁺ concentrations potassium is not accumulated by M. thermoautotrophicum via an electrogenic uniport mechanism.Non-common abbreviations TPP⁺ Tetra phenylphosphonium bromide - DTE Dithioerythritol - TCS 3,5,3,4-Tetrachlorosalycylanilide     

Effect of soil moisture and clipping stresses on the nutrient (N,P and K) concentration,uptake and use efficiency in one temperate and two tropical grasses

Summary The concentration, uptake and element use efficiency of N, P and K in one C₃ annual (Polypogon monspeliensis) and two C₄ (Echinochloa colonum, an annual, andDichathium annulatum, a perennial) grasses were determined during winter and summer seasons in monocultures raised in field plots at three moisture levels,viz. full, half and one-fourth of field capacity. At each moisture regime the plants were clipped thrice at moderate and severe levels corresponding to 40 and 80% of live green. The concentration of these elements was characteristic of the growth habit of these plants;e.g. the build up of concentration was maximum in leaf of the annuals while it was comparable in crown and leaf of Dichanthium. The N level was maximum in Polypogon. The nutrient use effiency was comparable in the two annuals and maximum K and N use were obtained in Polypogon and Dichanthium, respectively.     

The regulation of K+ influx in excised barley roots

The electrochemical potential differences for potassium, between excised barley (Hordeum vulgare L.) roots and external media containing 0.05 mM KCl+0.5 mM CaSO₄, were determined over a 4-h period during which initially low-K⁺ roots accumulated K⁺ by pretreatment in 50 mM KCl plus 0.5 mM CaCl₂. This pretreatment resulted in increased internal [K⁺], decreased K⁺ influx (as measured from 0.05 mM KCl+0.5 mM CaSO₄) and decreased values of . These observations indicate that the decline of K⁺ influx associated with increased internal K⁺ concentration cannot be accounted for by passive adjustment to the electrochemical gradient for this ion.     

The influence of potassium content on the kinetics of potassium influx into excised ryegrass and barley roots

The influx of K⁺ into excised roots of barley (Hordeum vulgare L.) and ryegrass (Lolium multiflorum L.) previously grown with or without K⁺ was measured in K⁺ solutions ranging in concentration from 0.01 to 50 mM. In both species the K⁺ influx was lower in the roots with high K⁺ content. The extent of reduction by high internal [K⁺] decreased with external concentration above 1 mM. These results support the contention that at high external concentrations passive diffusion makes significant contributions to observed fluxes.     

Effect of Quinine on Autoreceptor-Regulated Dopamine Release in the Rat Striatum

In vivo brain microdialysis was used to examine the role of potassium channel activation in dopamine (DA) autoreceptor function in the striatum of freely moving rats. Local application of the D2 receptor agonists quinpirole or N-0437 through the dialysis probe significantly reduced extracellular concentrations of DA. Local application of the D2 antagonist (-)-sulpiride produced significant increases in DA. Local perfusion with quinine, a K+ channel blocker, completely blocked the (-)-sulpiride-induced increases in DA but did not affect the DA agonist-induced decreases. (-)-Sulpiride completely blocked the effect of quinpirole on DA both in control and in quinine-treated animals. At the highest dose used, quinine caused a large transient increase in extracellular DA. Local application of tetrodotoxin or infusion of Mg2+ in the absence of Ca2+ did not prevent this quinine-induced transient increase in extracellular DA. These results demonstrate that DA autoreceptors in the striatum regulate DA release in awake, behaving animals. Local application of (-)-sulpiride increases DA levels by blocking the tonic activation of autoreceptors by endogenous DA. Quinine blocks the neuroleptic-induced increase in DA, perhaps by preventing the K+ channel opening that would normally accompany endogenous autoreceptor activation. The fact that exogenously applied DA receptor agonists can decrease extracellular DA levels in the presence of quinine suggests that they may be acting at extrasynaptic autoreceptors that are not tonically active in vivo. The effect of DA agonists on this site is via a DA receptor because it is blocked by (-)-sulpiride. However, this receptor does not appear to be coupled to a quinine-sensitive potassium channel.     

Soil beidellite and its relation to problems of potassium fertility and poor response to potassium fertilizers

Summary X-ray diffraction studies were made on soils with and without potassium fertility problems. All soils with clay fractions containing dominant beidellite or vermiculite showed potassium deficiency and lack of response to potassium fertilizer applications. All of the soils containing dominant montmorillonite or other clay minerals contained adequate potassium; on none of these, poor potassium response was reported. Special management practices are needed on the beidellitic and vermiculitic soils to increase potassium and ammonium fertilizer efficiency. Dominance of beidellite in the clay fraction should be reflected in soil classification. Establishment of a ‘beidellite’ family differentiating criterion in the Soil Taxonomy is proposed for this purpose.     

Effects of acid irrigation and liming on two clones of Norway spruce

Summary The effect of acid irrigation on the growth of rooted cuttings ofPicea abies (L.) Karst, was investigated in a pot experiment lasting 3 years. It involved two clones of Norway spruce, H 253 Bogstad I and H 254 Bogstad II. Irrigation water of pH 5.4, 4.0, 3.0 and 2.5 was used. Liming was included in the experiment.After the experimental period, the plants of all treatments were growing reasonably well. However, those plants irrigated at pH 2.5 were slightly discoloured. The plant mortality was only 3% throughout the experiment, and was not connected to acid irrigation. The limiting growth factor was N. All other nutrient elements measured in the plants were close to optimal concentration. Plants irrigated at pH 2.5, and to some extent at pH 3, contained excessively high concentrations of Al, t-S and SO₄. The total amount of Ca, Fe and Mn taken up by the plants decreased with increasing soil acidity. The increased growth of clone H 254 relative to H 253, produces a corresponding impression on soil characteristics. Soil acidity is governed by acid irrigation and CaCO₃ application, but the clonal effects are also of importance. Norway spruce appears to be tolerant to Al concentrations as high as 50 mmol/kg in the needles.