Acute Effect of Ammonia on Branched-Chain Amino Acid Oxidation and Incorporation into Proteins in Astrocytes and in Neurons in Primary Cultures

14CO2 production and incorporation of label into proteins from the labeled branched-chain amino acids, leucine, valine, and isoleucine, were determined in primary cultures of neurons and of undifferentiated and differentiated astrocytes from mouse cerebral cortex in the absence and presence of 3 mM ammonium chloride. Production of 14CO2 from [1-14C]leucine and [1-14C]valine was larger than 14CO2 production from [U-14C]leucine and [U-14C]valine in both astrocytes and neurons. In most cases more 14CO2 was produced in astrocytes than in neurons. Incorporation of labeled branched-chain amino acids into proteins varied with the cell type and with the amino acid. Addition of 3 mM ammonium chloride greatly suppressed 14CO2 production from [1-14C]-labeled branched chain amino acids but had little effect on 14CO2 production from [U-14C]-labeled branched-chain amino acids in astrocytes. Ammonium ion, at this concentration, suppressed the incorporation of label from all three branched-chain amino acids into proteins of astrocytes. In contrast, ammonium ion had very little effect on the metabolism (oxidation and incorporation into proteins) of these amino acids in neurons. The possible implications of these findings are discussed, especially regarding whether they signify variations in metabolic fluxes and/or in magnitudes of precursor pools.     

Local movements and population size of European eels,Anguilla anguilla,in a small lake in southwestern Spain

Synopsis Radio telemetry was used to study the movements of European eels,Anguilla anguilla, in a small (1.2 ha) lake in southwestern Spain in March and April, 1985. Observations were taken on the locations of 7 eels at least once each 2 h for a combined total of 1713 h. The size of individual activity regions varied from 2700 to 1300 m². Eels covered a larger area at night than during the day, with an average of 23% and 42% of the activity region used during the day and night respectively. Average distance moved between observations was significantly greater at night than in the day. Eels tracked during rainy and cloudy weather were more active during the day and used a larger total area than did those tracked during drier, more stable weather. The standing crop of eels was estimated to be about 77 kg ha^–1.     

Twenty-four-hour variations in activity,core temperature,metabolic rate,and respiratory quotient in captive chinese pangolins

The circadian rhythms in activity, core temperature (T_c), O₂ consumption, CO₂ production, and respiratory quotient (RQ) were monitored in four captive Chinese pangolins (Manis pentadactyla). The pangolins were strictly nocturnal, never emerging from their nest before 1600 h, and their intermittent activity continued no later than 0230. As is usual in nocturnal mammals, the highest values observed in T_c, O₂ consumption, and CO₂ production occurred during the night; the lowest values occurred during the day. The magnitude of the variation in T_c, O₂ consumption, CO₂ production, and RQ averaged 1.2°C, 1.3 ml O₂ kg^?1 min^?1, 1.2 ml CO₂ kg^?1 min^?1, and 0.24, respectively. The circadian pattern in RQ was independent of activity, T_c, and the metabolic parameters and was of a different character than the patterns exhibited in the other variables. RQ remained constant at either a high or low level for long periods (8–10 h) and then increased or decreased relatively rapidly (1–2h) to the other level as in a square wave, whereas the rhythms in the other variables are similar to sine waves. The sharp increase in RQ was followed by a slow decline in T_c, and the sharp decline in RQ was followed by a slow increase in T_c.     

Succinate metabolism related to lipid synthesis in the housefly Musca domestica L

The metabolism of succinate was examined in the housefly Musca domestica L. The labeled carbons from [2,3-¹⁴C]succinate were readily incorporated into cuticular hydrocarbon and internal lipid, whereas radioactivity from [1,4-¹⁴C]succinate was not incorporated into either fraction. Examination of the incorporation of [2,3-¹⁴C]succinate, [1-¹⁴C]acetate, and [U-¹⁴C]proline into hydrocarbon by radio-gas-liquid chromatography showed that each substrate gave a similar labeling pattern, which suggested that succinate and proline were converted to acetyl-CoA prior to incorporation into hydrocarbons. Carbon-13 nuclear magnetic resonance showed that the labeled carbons from [2,3-¹³C]succinate enriched carbons 1, 2, and 3 of hydrocarbons with carbon-carbon coupling showing that carbons 2 and 3 of succinate were incorporated as an intact unit. Radio-high-performance liquid chromatographic analysis of [2,3-¹⁴C]succinate metabolism by mitochondrial preparations showed that in addition to labeling fumarate, malate, and citrate, considerable radioactivity was also present in the acetate fraction. The data show that succinate was not converted to methylmalonate and did not label hydrocarbon via a methylmalonyl derivative. Malic enzyme was assayed in sonicated mitochondria prepared from the abdomens and thoraces of 1- and 4-day-old insects; higher activity was obtained with NAD⁺ in mitochondria prepared from thoraces, whereas NADP⁺ gave higher activity with abdomen preparations. These data document the metabolism of succinate to acetyl-CoA and not to a methylmalonyl unit prior to incorporation into lipid in the housefly and establish the role of the malic enzyme in this process.     

The metabolism of deoxyguanosine in mitochondria. Characterization of the uptake process

Summary The uptake of deoxyguanosine by rat liver mitochondria was characterized. The process required an intact mitochondrial membrane and exhibited a dependence on added phosphate. Deoxyguanosine uptake was minimally influenced by Mg²⁺ or Mn²⁺, but Ca²⁺ at concentrations above 0.5 mM were detrimental. Of the deoxynucleosides tested, only deoxyinosine inhibited the uptake of deoxyguanosine. The ribonucleoside guanosine was not observed to compete with its deoxynucleoside analog. Known inhibitors of nucleoside transport, cytochalasin B and NBMPR, did not block deoxyguanosine uptake, but the sulfhydryl reagents NEM and pCMB were both inhibitory. The uptake of deoxyguanosine was shown to be a saturable process and an apparent Km of 0.64 M was calculated from a Hanes plot.     

Glucose and amino acid metabolism in chick telencephalon slices: Changes with incubation conditions and animals' development

Glucose and amino acid metabolism in 1- and 30-day-old chick telencephalon slices was studied in two incubation media in the presence or in the absence of a continuous oxygenation. Medium 1 has a composition and a tonicity similar to cerebrospinal fluid, medium 2 is hypertonic and does not contain any K⁺ ions. The incorporation of glucose carbon into amino acids and the distribution of radioactivity between the different amino acids are close to the ones observed in the chick brain in vivo only when the slices are incubated in medium 1, with oxygen at 30 days and without oxygen for the 1-day-old chick. It also appears that if oxygenation is necessary for incubation of mature brain tissue in vitro, the absence of the medium oxygenation is more suitable for the study of glucose metabolism in 1-day-old chick brain slices.     

Ecological aspects of thermoregulation at high altitudes: the case of andean Liolaemus lizards in northern Chile

Summary We document activity field temperatures, daily activity patterns, and extent of thermoregulation in four species of Liolaemus lizards inhabiting at high altitude (above 3500 m) in the Andes of northern Chile. These four species have similar activity field temperature (Tb near 29°C) despite their being distributed at different altitudinal belts. However, conspicuous differences exist between higher-altitude (L. alticolor and L. jamesi) and lower-altitude (L. islugensis and L. ornatus) lizards regarding extent of thermoregulation and activity period. Some differences in morphology, behavior, and patterns of microhabitat occupancy are also apparent among these four species and are seemingly related to the thermal environment to which they are subjected. In comparison to eight low-altitude Liolaemus species in central Chile (Tb near 35°C) the four high-altitude species in northern Chile have lower activity field temperature. The latter is apparently due to the constraints imposed by the harsh Andean thermal environment, a hypothesis supported by the fact that high-altitude Liolaemus lizards under laboratory conditions demonstrate body temperatures that exceed by 5°C or more, those recorded in the field.     

Putrescine metabolism in excised cotyledons of Pinus radiata cultured in vitro

The metabolism of ¹⁴C-putrescine and the changes in the endogenous concentrations of putrescine, spermidine and spermine were studied when cotyledons of Pinus radiata D. Don were cultured under shoot-forming (SF, + N⁶-benzyladenine) and non-shoot-forming (NSF, - N⁶-benzyladenine) conditions. Differences in the total uptake of ¹⁴C-putrescine during a 2 h pulse feeding were not significant between the SF and NSF cotyledons except on day 3. The maximum uptake of label was on day 3 in the SF cotyledons, which released the highest amount of ¹⁴CO₂ as well. ¹⁴C from the labeled putrescine was incorporated mainly into γ-aminobutyric acid, aspartate and glutamate. High performance liquid chromatography of the endogenous polyamines indicated that spermidine was the most predominant polyamine in the cultured cotyledons of radiata pine. Spermine increased by about 60% in the SF and 25% in the NSF cotyledons between days 0 and 3 of culture.     

Nuclear-magnetic-resonance imaging of leaves ofMesembryanthemum crystallinum L. plants grown at high salinity

Differences in water binding were measured in the leaf cells ofMesembryanthemum crystallinum L. plants grown under high-salinity conditions by using nuclear-magnetic-resonance (NMR) imaging. The 7-Tesla proton NMR imaging system yielded a spatial resolution of 20·20·100 m³. Images recorded with different spin-echo times (4.4 ms to 18 ms) showed that the water concentrations in the bladder cells (located on the upper and lower leaf surface), in the mesophyll cells and in the water-conducting vessels were nearly identical. All of the water in the bladder cells and in the water-conducting vessels was found to be mobile, whilst part of the water in the mesophyll cells was bound. Patches of mesophyll cells could be identified which bound water more strongly than the surrounding mesophyll cells. Optical investigations of leaf cross-sections revealed two types of mesophyll cells of different sizes and chloroplast contents. It is therefore likely that in the small-sized mesophyll cells water is strongly bound. A long-term asymmetric water exchange between the mesophyll cells and the bladder cells during Crassulacean acid metabolism has been described in the literature. The high density of these mesophyll cells in the lower epidermis is a possible cause of this asymmetry.Abbreviations CAM Crassulacean acid metabolism - NMR nuclear magnetic resonance - T_E spin-echo time     

Water storage and osmotic pressure influences on the water relations of a dicotyledonous desert succulent

Abstract Water storage and nocturnal increases in osmotic pressure affect the water relations of the desert succulent Ferocactus acanthodes, which was studied using an electrical circuit analog based on the anatomy and morphology of a representative individual. Transpiration rates and osmotic pressures over a 24-h period were used as input variables. The model predicted water potential, turgor pressure and water flow for various tissues. Plant capacitances, storage resistances and nocturnal increases in osmotic pressure were varied to determine their role in the water relations of this dicotyledonous succulent. Water coming from storage tissues contributed about one-third of the water transpired at night: the majority of this water came from the nonphotosynthetic, water storage parenchyma of the stem. Time lags of 4 h were predicted between maximum transpiration and maximum water uptake from the soil. Varying the capacitance of the plant caused proportional changes in osmotically driven water movement but changes in storage resistance had only minor effects. Turgor pressure in the chlorenchyma depended on osmotic pressure, but was fairly insensitive to doubling or halving of the capacitance or storage resistance of the plant. Water uptake from the soil was only slightly affected by osmotic pressure changes in the chlorenchyma. For this stem succulent, the movement of water from the chlorenchyma to the xylem and the internal redistribution of water among stem tissues were dominated by nocturnal changes in chlorenchyma osmotic pressure, not by transpiration.