Fluorometric estimation of surface potential change associated with chemotactic stimulation in Tetrahymena pyriformis

For the quantitative estimation of surface potential change in intact cells a method was devised with the use of fluorescent probes, 8-anilino-1-naphthalenesulfonate (ANS) and N-phenyl-1-naphthylamine (NPN). Estimated values in liposomes were compared with changes in the zeta potential determined from electrophoresis. Both values agreed within the experimental variation, showing the usefulness of the method. The method was also applied to Tetrahymena pyriformis, which exhibits chemotaxis to various chemical stimuli. The surface potential change was observed when the cell was stimulated not only by inorganic salts but also by electrically neutral, hydrophobic compounds. The surface potential started to change in accordance with the depolarization of the membrane potential, except for the case of K⁺. Changes in the surface potential of T. pyriformis in response to Ca²⁺ and K⁺ were compared with those in the membrane potential. The quantitative contribution of the surface potential to cell depolarization associated with chemoreception is discussed.     

Seasonal fluctuations of the mineral concentration of alder (Alnus glutinosa (L.) Gaertn.) from the field

Summary The seasonal fluctuation of N, P, K, Ca, Mg, Fe, Mn, Mo, and Co, in leaves, roots and nodules of 40–50 year oldAlnus glutinosa trees growing at four different locations along the banks of the Tormes river, in the province of Salamanca, was studied. Also, the evolution of the soil organic matter under the trees sampled was evaluated. The data obtained for the various nutrient elements in the three plant parts are statistically treated at the significance levels of 99–95 per cent, and some remarks as to the nutritional status of the European alder in respect to the nutrients and its contribution to soil nutrient-cycling are provided. A positive correlation was found between N–P, N–K, N–Mg, and N–Mo, in leaves, and between N–P, N–K, N–Fe, N–Mn, and N–Mo in root nodules. In roots only, no significance at any level was obtained between N and any of the elements analyzed.     

Movement of sodium into human platelets

Addition of ADP induces platelets in plasma to undergo shape change from a disc to a spiny sphere and to develop adhesiveness, i.e. to aggregate. The aggregation of human platelets by ADP is associated with a net uptake of Na⁺. The present experiments demonstrate that the induction of shape change by ADP in acidified or EGTA-treated plasma conditions which inhibit aggregation, is also associated with a movement of Na⁺ into platelets. When ADP-induced platelet shape change and aggregation is inhibited by prostaglandin E₁ Na⁺ uptake is also blocked. Platelets aggregated by epinephrine do not take up Na⁺. In a manner analogous to the effect of ADP, polylysine also induces Na⁺ uptake during aggregation. Vasopressin, in a manner analogous to epinephrine, induces aggregation without Na⁺ uptake. The increase in platelet Na⁺ resulting from ouabain inhibition of Na⁺ efflux induces an increase in the aggregation response to ADP and to epinephrine.     

Changes in the fine structure of the testicular Leydig cells of the seasonally-breeding bat,Myotis adversus

Summary Leydig cells of the bat, Myotis adversus, have been examined by electron microscopy throughout fourteen months. During the breeding season the Leydig cells become hypertrophied and are characterised by prominent areas of agranular endoplasmic reticulum and numerous small, membrane-bound granules. Microperoxisomes are also observed. During the period of testicular regression. Leydig cell size and the number of membrane-bound granules are greatly reduced. Lipid droplets and dense bodies are more numerous.     

Acclimation to temperature and death at changing lethal temperatures in the freshwater fish Chalcalburnues chalcoides

1. 1.|In the freshwater fish Chalcalburnus chalcoides, an increase in the body (standard) size caused decreases in the upper LT-50 from 36.6° to 36.0°C and lower LT-50 from 6.3° to 5.3°C

2. 2.|The fish acclimated to constant temperatures between 10°C and 30°C showed reasonable heat acclimation and also reasonable cold acclimation. Thus, an increase in the acclimation temperature from 10°C to 30°C caused increases in the upper LT-50 from 34° to 36.2°C and the lower LT-50 from 1.25 to 6.5°C.

3. 3|The mean survival time — temperature curves of 10°, 20° and 30°C acclimated fish at various constant temperatures showed decreased in the survival tim ewith increasing lethal temperatures. Furthermore, an increase in the acclimation temperature causes a shift in the survival duration-temperature curve to the right, i.e., the fish become more heat resistant. Thus, the mean survival duration of 10°, 20° and 30°C acclimated fish at 35°C were 7.5, 79.6 and 530 minutes, respectively.

4. 4.|The effect of the thermal experience to changing lethal temperatures depends on the first lethal temperature to which the fish were exposed as well as the sequence of temperature changes. In the experiments in which the first lethal temperatures were between 32° and 34°C and the temperature was varied in an ascending order, their thermal resistance was increased and the fish required 114 to 174% of the expected lethal doses to die while in the experiments in which the starting temperature were between 38° and 40°C and the temperature varied in descending order, the fish become more sensitive to the upper lethal temperature and they died after receiving only 62 to 81% of the expected lethal doses. Thus, with a gradual increase in the lethal temperature, the fish show additional acclimation in the zone of resistance which in turn causes an increase in the thermal resistance. This may have ecological significance in nature.

Fish community structure in lakeshore lagoons on long point,lake Erie,Canada

Synopsis The interaction of fluctuating water level and the Long Point topography is discussed with regard to the formation of lake-shore lagoons and their subsequent invasion by fishes. The species composition of fish communities in these lagoons was determined by 12 collections from lagoons (23 m² to 25 ha) which yielded 33 fish species. The relationship of the number of fish species to lagoon area was lognormal; that is, with increasing lagoon area the number of species rose rapidly at first but soon decreased sharply, probably limited by the number of species capable of surviving in the habitat. The marked effect of area on species number in small lagoons is considered in the context of a hypothetical seasonal pattern of occasional invasion and continuous extinction.A sequence of reproductive guilds was found. In beach lagoons psammophils and lithophils were dominant. In a large vegetated lagoon in the interior phytophils and speleophils were dominant. This sequence was paralleled by an increase in the proportion of guarding and nesting species in the communities. The sequence was disrupted in the small interior lagoons where adaptations for survival were more important than reproductive strategy.     

Sulphydryl groups of (Na+ + K+)-ATPase from rectal glands of Squalus acanthias: Detection of ligand-induced conformational changes

1. Modification of the Class II sulphydryl groups on the (Na⁺ + K⁺)-ATPase from rectal glands of Squalus acanthias with N-ethylmaleimide has been used to detect conformational changes in the protein. The rates of inactivation of the enzyme and the incorporation of N-ethylmaleimide depend on the ligands present in the incubation medium. With 150 mM K⁺ the rate of inactivation is largest (k₁ = 1.73 mM^?1 · min^?1) and four SH groups per α-subunit are modified. The rate of inactivation in the presence of 150 mM Na⁺ is smaller (k₁ = 1.08 mM^?1 · min^-1) but the incorporation of N-ethylmaleimide is the same as with K⁺. 2. ATP in micromolar concentrations protects the Class II groups in the presence of Na⁺ (k₁ = 0.08 mM^?1 · min^?1 at saturating ATP) and the incorporation id drastically reduced. ATP in millimolar concentrations protects the Class II groups partially in the presence of K⁺ (k₁ = 1.08 mM^?1 · min^?1) and three SH groups are labelled per α subunit. 3. The K⁺ -dependent phosphatase is inhibited in parallel to the (Na⁺ + K⁺)-ATPase under all conditions, and the ligand-dependent incorporation of N-ethylmaleimide was on the α-subunit only. 4. It is shown that the difference between the Na⁺ and K⁺ conformations sensed with N-ethylmaleimide depends on the pH of the incubation medium. At pH 6 there is a very small difference between the rates of inactivation in the presence of Na⁺ and K⁺, but at higher pH the difference increases. It is also shown that the rate of inactivation has a minimum at pH 6.9, which suggests that the conformation of the enzyme changes with pH. 5. Modification of the Class III groups with N-ethylmaleimide-whereby the enzyme activity is reduced from about 16% to zero-shows that these groups are also sensitive to conformational changes. As with the Class II groups, ATP in micromolar concentrations protects in the presence of Na⁺ relative to Na⁺ or K⁺ alone. ATP in millimolar concentrations with K⁺ present increases the rate of inactivation relative to K⁺ alone, in contrast to the effect on the Class II groups. 6. Modification of the Class II groups with a maleimide spin label shows a difference between Class II groups labelled in the presence of Na⁺ (or K⁺) and Class II groups labelled in the presence of K + ATP, in agreement with the difference in incorporation of N-ethylmaleimide. The spectra suggest that the SH group protected by ATP in the presence of K⁺ is buried in the protein. 7. The results suggest that at least four different conformations of the (Na⁺ + K⁺)-ATPase can be sensed with N-ethylmaleimide: (i) a Na⁺ form of the enzyme with ATP bound to a high-affinity site (E₁-Na-ATP); (ii) a Na⁺ form without ATP bound (E₁-Na); (iii) a K⁺ form without ATP bound (E₂-K); and (iv) an enzyme form with ATP bound to a low-affinity site in the presence of K⁺, probably and E₁-K-ATP form.     

Electron paramagnetic resonance study on calmodulin: conformational change and interaction with divalent cations

Biologically active spin labelled derivatives of calmodulin were prepared and used to study CA²⁺- and Mg²⁺-induced conformational changes of the protein. The rotational correlation time of the spin labelled residues increased upon addition of divalent cations. Two calcium ions per spin labelled calmodulin were found to induce a 75% conformational change, whereas four calcium ions were necessary for a maximum conformational change. The increase in rotational correlation time induced by Mg²⁺ is less pronounced. Two different covalently attached spin labels (iodoacetamide and maleimide) were compared and marked differences were found in their chemical stability. The binding of manganese ions to calmodulin could be observed directly from the electron paramagnetic resonance spectra of these paramagnetic ions. Two specific classes of binding sites, each binding two manganese ions with $\text{k}{}_{\mathrm{<mtext>D</mtext>}}\text{= 0.6 × 10}{}^{\mathrm{?6}}\text{m}\text{and}\text{k}{}_{\mathrm{D}}\text{= 3 × 10}{}^{\mathrm{?5}}\text{m}$, respectively, were determined. Further ion binding occurs at non-specific sites.     

Neuroendocrine effects of light

The light/dark cycle to which animals, and possibly humans, are exposed has a major impact on their physiology. The mechanisms whereby specific tissues respond to the light/dark cycle involve the pineal hormone melatonin. The pineal gland, an end organ of the visual system in mammals, produces the hormone melatonin only at night, at which time it is released into the blood. The duration of elevated nightly melatonin provides every tissue with information about the time of day and time of year (in animals that are kept under naturally changing photoperiods). Besides its release in a circadian mode, melatonin is also discharged in a pulsatile manner; the physiological significance, if any, of pulsatile melatonin release remains unknown. The exposure of animals including man to light at night rapidly depresses pineal melatonin synthesis and, therefore, blood melatonin levels drop precipitously. The brightness of light at night required to depress melatonin production is highly species specific. In general, the pineal gland of nocturnally active mammals, which possess rod-dominated retinas, is more sensitive to inhibition by light than is the pineal gland of diurnally active animals (with cone-dominated retinas). Because of the ability of the light/dark cycle to determine melatonin production, the photoperiod is capable of influencing the function of a variety of endocrine and non-endocrine organs. Indeed, melatonin is a ubiquitously acting pineal hormone with its effects on the neuroendocrine system having been most thoroughly investigated. Thus, in nonhuman photoperiodic mammals melatonin regulates seasonal reproduction; in humans also, the indole has been implicated in the control of reproductive physiology.Summary of a Plenary Lecture presented by the author in Vienna, August, 1990