The effect of acclimation temperature on the activation energies of state III respiration and on the unsaturation of membrane lipids of goldfish mitochondria.

The influence of the acclimation temperature on the thermotropic behaviour of mitochondrial respiration and on the degree of unsaturation of mitochondrial membrane lipids has been studied. The mitochondria were isolated from red muscle, white muscle and liver of goldfish acclimated to 5, 20 and 30 degrees C. ADP-activated succinate oxidation was measured at different temperatures and resulted in non-linear Arrhenius-plots with breaks between 10 and 23 degrees C. As for the break-temperatures, there was found a shift downwards in preparations of decreased acclimation temperatures. This could be caused by a changed composition of membrane lipids and a simultaneous shift of the membrane phase transition temperature. Therefore, the fatty acid composition of all membrane preparations was analyzed. However, no consistent change of the degree of unsaturation due to a changed acclimation temperature could be found.     

Thermal adaptation of Tetrahymena membranes with special reference to mitochondria Role of cardiolipin in fluidity of mitochondrial membranes

During temperature acclimation of Tetrahymena pyriformis, the changes in fluidity and composition of total lipids from three membrane fractions, mitochondria, pellicles and microsomes were studied by a spin-label technique using a stearate probe and thin-layer and gas-liquid chromatography. The increase of fluidity observed in microsomal and pellicular lipids following the temperature shift from 39 to 15°C corresponds with the increase of the ratio of total unsaturated to saturated fatty acid content. However, despite the increase of this ratio, the fluidity of mitochondrial lipids was found to be constant up to 10 h after the temperature shift. The fluidity of total lipids of mitochondria isolated from Tetrahymena cells grown at 39°C was not changed by removal of cardiolipin, whereas cardiolipin-depleted lipids of mitochondria from 15°C-acclimated cells showed a decrease in fluidity. The re-addition of cardiolipin to the mitochondrial lipids depleted of cardiolipin restored the fluidity to the initial leve, thereby confirming the rigidifying effect of cardiolipin in cold-acclimated cells. These results suggest that cardiolipin may be implicated in maintaining consistent fluidity of mitochondrial membranes against change in thermal environment.     

A steady state and differential polarised phase fluorimetric study of the liver microsomal and mitochondrial membranes of thermally acclimated green sunfish (Lepomis cyanellus)

The liver mitochondrial and microsomal membranes of green sunfish and rat were examined by steady state polarisation and differential polarised phase fluorimetry to determine the effects of seasonal adaptation of membrane dynamic structure to temperature. Steady state polarisation studies indicated that the liver mitochondria of green sunfish acclimated to different temperatures showed a greater partial compensation of membrane fluidity for the altered acclimation temperature than did liver microsomal membranes. The fatty acid composition of both membrane preparations generally became more unsaturated at lower acclimation temperatures, though the differences between 5°C and 25°C acclimated fish were more pronounced in the mitochondrial fraction than in the microsomal fraction.Differential polarised phase fluorimetric studies indicated that the rotations of diphenylhexatriene in mitochondrial and microsomal membranes were highly hindered, though the hindrance offered by membranes of 25°C acclimated green sunfish was far greater than that offered by the membranes of 5°C acclimated fish, thus supporting the concept of homeoviscous adaptation. The absolute rotational rate was not consistently affected by acclimation treatment.     

Influence of environmental temperature on mitochondrial membranes

Mitochondrial phospholipids from goldfish lateral line muscle were analysed with respect to polar and apolar groups. Groups of 20 goldfish, acclimated to 5, 20 and 30°C, were used. Temperature-induced shifts of both polar and apolar groups of the mitochondrial phospholipids were observed. The fatty acid composition of mitochondrial phospholipids is characterized by a large amount of polyenoic acids, dominated by docosahexaenoic acid and by octadecadienoic acid. At the higher acclimation temperatures, a significant decrease in docosahexaenoic acid is found. However, the resultant effect of environmental temperature on the degree of unsaturation is small, in contrast to the marked effect on mean chain length. Pronounced changes in the molar ratio of phosphatidylcholine and phosphatidylethanolamine are seen; a decrease in mitochondrial phosphatidylcholine is observed at low acclimation temperature, which is compensated for by a nearly equal increase in phosphatidylethanolamine. The main phospholipids are, apparently, phosphatidylcholine, phosphatidylethanolamine and cardiolipin, comprising 90% of the total pool of 12 species. It is found that the anionic nature of the phospholipids is increased at low acclimation temperatures. We discuss this effect and its probable importance in the stabilization of the surface potential of the mitochondrial membranes.     

Hormones and liver mitochondria: Influence of growth hormone,thyroxine, testosterone,and insulin on thermotropic effects of respiration and fatty acid composition of membranes

The effects of hypophysectomy and subsequent administration of growth hormone, thyroxine, insulin, and testosterone were examined in rat liver for the relationship between the thermotropic effects on State 3 respiration (ADP induced) and fatty acid composition of the phospholipid fraction of intact mitochondria as well as of inner membrane vesicles. The Arrhenius profile for energy-linked (succinate) State 3 respiration of mitochondria from hypophysectomized rats lacked the discontinuity at 23.5 °C seen with mitochondria from normal rats. After injections of the hormones the discontinuity representing the transition temperature from gel to liquid crystalline state of lipids occurred at different temperatures: 18.5 °C for growth hormone, 26.0 °C for thyroxine, 19.5 °C for growth hormone + thyroxine, 27.6 °C for insulin, and 25.3 °C for testosterone. The energy of activation between 37.5 and 23.5 °C was 1.9 times greater for hypophysectomy than for controls. Growth hormone was the most effective in restoring the energy of activation to normal, above as well as below transition temperature. The effect of thyroxine appears to be due to a larger stimulation of the State 4 respiration than that of growth hormone, insulin, or testosterone, especially at higher temperatures. Phospholipids extracted from intact mitochondria or inner membrane vesicles of hypophysectomized rats contained less arachidonic acid (20:4) and more linoleic acid (18:2) than those of normal rats. In addition, the contents of some of the minor fatty acids were also changed. Calculated unsaturation index showed an 18.8 and 14.9% depletion in unsaturation in whole mitochondria and inner membranes, respectively. Among the different hormones used to treat the hypophysectomized rats, growth hormone was the most effective in restoring the transition temperature and fatty acid composition to normal levels and increasing the gain in body weight. Although the other hormones increased total unsaturation index to some extent, some of the individual fatty acids were affected differently. Good correlation exists between the unsaturation index of mitochondrial fatty acids and transition temperature of State 3 respiration. These results strongly suggest a role for the hormones, particularly growth hormone, in the control of mitochondrial membrane fluidity of hypophysectomized rat liver, through fatty acid composition of phospholipids.     

Temperature-induced specific lipid desaturation in the thermophilic cyanobacterium Synechococcus vulcanus

Cyanobacteria desaturate fatty acids in the membrane lipids in response to decrease in temperature. We examined the changes in lipid and fatty acid composition in the thermophilic cyanobacterium Synechococcus vulcanus, which is characterized by an optimum growth temperature of 55°C. During temperature acclimation to 45°C or 35°C, the cells synthesized oleic acid at the expense of stearic acid in the membrane lipids. Unlike mesophilic cyanobacteria, S. vulcanus did not show any significant adaptive desaturation in the galactolipids monogalactosyl diacylglycerol and digalactosyl diacylglycerol, that comprise 50% and 30% of total membrane lipids, respectively. The major changes in fatty acid unsaturation were observed in the sulfolipid sulfoquinovosyl diacylglycerol.     

Correlations between the thermal stability of chloroplast (thylakoid) membranes and the composition and fluidity of their polar lipids upon acclimation of the higher plant,Nerium oleander,to growth temperature

The thermal stability of excitation transfer from pigment proteins to the Photosystem II reaction center of Nerium oleander adjusts by 10 Celsius degrees when cloned plants grown at 20°C/15°C, day/night growth temperatures are shifted to 45°C/32°C growth temperature or vice versa. Concomitant with this adjustment is a decrease in the fluidity of thylakoid membrane polar lipids as determined by spin labeling. The results are consistent with the hypothesis that there is a limiting maximum fluidity compatible with maintenance of native membrane structure and function. This limiting fluidity was about the same as for a number of other species which exhibit a range of thermal stabilities. Inversely correlated shifts in lipid fluidity and thermal stability occurred during the time course of acclimation of N. oleander to new growth temperatures. Thus, the temperature at which the limiting fluidity was reached changed during acclimation while the limiting fluidity remained constant. Although the relative proportion of the major classes of membrane polar lipids remained constant during adjustments in fluidity, large changes occured in the abundance of specific fatty acids. These changes were different for the phospho- and galacto-lipids suggesting that the fatty acid composition of these two lipid classes is regulated by different mechanisms. Comparisons between membrane lipid fluidity and fatty acid composition indicate that fluidity is not a simple linear function of fatty acid composition.     

Thermal adaptation of Tetrahymena membranes with special reference to mitochondria. Role of cardiolipin in fluidity of mitochondrial membranes

During temperature acclimation of Tetrahymena pyriformis, the changes in fluidity and composition of total lipids from three membrane fractions, mitochondria, pellicles and microsomes were studied by a spin-label technique using a stearate probe and thin-layer and gas-liquid chromatography. The increase of fluidity observed in microsomal and pellicular lipids following the temperature shift from 39 to 15 degrees C corresponds with the increase of the ratio of total unsaturated to saturated fatty acid content. However, despite the increase of this ratio, the fluidity of mitochondrial lipids was found to be constant up to 10 h after the temperature shift. The fluidity of total lipids of mitochondria isolated from Tetrahymena cells grown at 39 degrees C was not changed by removal of cardiolipin, whereas cardiolipin-depleted lipids of mitochondria from 15 degrees C-acclimated cells showed a decrease in fluidity. The re-addition of cardiolipin to the mitochondrial lipids depleted of cardiolipin restored the fluidity to the initial level, thereby confirming the rigidifying effect of cardiolipin in cold-acclimated cells. These results suggest that cardiolipin may be implicated in maintaining consistent fluidity of mitochondrial membranes against change in thermal environment.     

Thermal adaptation of Tetrahymena membranes with special reference to mitochondria II. Preferential interaction of cardiolipin with specific molecular species of phospholipid

A specific effect of cardiolipin on fluidity of mitochondrial membranes was demonstrated in Tetrahymena cells acclimated to a lower temperature in the previous report (Yamauchi, T., Ohki, K., Maruyama, H. and Nozawa, Y. (1981) Biochim. Biophys. Acta 649, 385–392). This study was further confirmed by the experiment using fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH). Anisotropy of DPH for microsomal and pellicular total lipids from Tetrahymena cells showed that membrane fluidity of these lipids increased gradually as the cells were incubated at 15°C after the shift down of growth temperature from 39°C. However, membrane fluidity of mitochondrial total lipids was kept constant up to 10 h. This finding is compatible with the result obtained using spin probe in the previous report. Additionally, the break-point temperature of DPH anisotropy was not changed in mitochondrial lipids whereas those temperatures in pellicular and microsomal lipids lowered during the incubation at 15°C. Interaction between cardiolipins and various phospholipids, which were isolated from Tetrahymena cells grown at 39°C or 15°C and synthesized chemically, was investigated extensively using a spin labeling technique. The addition of cardiolipins from Tetrahymena cells grown at either 39°C or 15°C did not change the membrane fluidity (measured at 15°C) of phosphatidylcholine from whole cells grown at 39°C. On the other hand, both cardiolipins of 39°C-grown and 15°C-grown cells decreased the membrane fluidity of phosphatidylcholine from Tetrahymena cells grown at 15°C. The same results were obtained for phosphatidylcholines of mitochondria and microsomes. Membrane fluidity of phosphatidylethanolamine, isolated from cells grown at 15°C, was reduced to a small extent by Tetrahymena cardiolipin whereas that of 39°C-grown cells was not changed. Representative molecular species of phosphatidylcholines of cells grown at 39°C and 15°C were synthesized chemically; 1-palmitoyl-2-oleoylphosphatidylcholine for 39°C-grown cells and dipalmitoleoylphosphatidylcholine for 15°C-grown ones. By the addition of Tetrahymena cardiolipin, the membrane fluidity of 1-palmitoyl-2-oleoylphosphatidylcholine was not changed but that of dipalmitoleoylphosphatidylcholine was decreased markedly. These phenomena were caused by Tetrahymena cardiolipin. However, bovine heart cardiolipin, which has a different composition of fatty acyl chains from the Tetrahymena one, exerted only a small effect.     

Temperature-induced changes in fatty acid dynamics of the intertidal grazer Platychelipus littoralis (Crustacea,Copepoda, Harpacticoida): Insights from a short-term feeding experiment

Dietary lipids, and in particular the essential fatty acids (EFA), EPA (20:5ω3) and DHA (22:6ω3), guarantee the well-being of animals and are recognized for their potential bottom-up control on animal populations. They are introduced in marine ecosystems through primary producers and when grazed upon, they are consumed, incorporated or modified by first-level consumers. As the availability of EFA in the ecosystem is affected by ambient temperature, the predicted rise in ocean temperature might alter the availability of these EFA at the basis of marine food webs. Despite the FA bioconversion capacity of certain benthic copepod species, their lipid (FA) response to varying temperatures is understudied. Therefore, the temperate, intertidal copepod Platychelipus littoralis was offered a mono and mixed diatom diet at 4, 15 °C (normal range) and at 24 °C (elevated temperature) to investigate the combined effects of temperature and resource availability on its FA content and composition. P. littoralis showed a flexible thermal acclimation response. Cold exposure increased the degree of FA unsaturation and the EPA%, and induced a shift towards shorter chain FA in the copepod’s membranes. Furthermore, a mixed diet reduced the impact of heat stress on the copepod’s membrane FA composition. Temperature affected the trophic transfer of EPA and DHA differently. While dietary resources could fully compensate for the temperature effects on total lipid and EPA content in the copepods, no such counterweigh was observed for the DHA dynamics. Heat stress lowered the DHA concentration in copepods regardless of the resources available and this implies negative effects for higher trophic levels.     

Change in lipid composition in eastern oyster (Crassostrea virginica Gmelin) exposed to constant or fluctuating temperature regimes

A temperature decrease usually induces an ordering effect in membrane phospholipids that can lead to membrane dysfunction. Ectotherms typically counteract this temperature effect by remodeling membrane lipids as stipulated in the homeoviscous adaptation theory (HVA). Previous studies mostly focused on the remodeling of membrane lipids during long-term acclimatization or acclimation at constant temperature regimes, whereas in nature, many organisms experience variations in temperature on a daily basis and must react to this changing thermal environment. The objective of this study was to examine the composition of membrane lipids in oysters subjected to long-term acclimation at constant temperatures (12 or 25 degrees C) or to environmentally realistic daily fluctuations in temperature between 12 and 25 degrees C for 7 d. The lipid composition of gill in oysters subjected to long-term acclimation at a constant temperature or to daily temperature fluctuations varied in a way consistent with HVA: oysters adjusted their phospholipid to sterol ratio in response to long-term acclimation to a constant temperature but not to daily temperature fluctuations. In contrast, the unsaturation index of polar lipids in oysters varied in response to both long-term acclimation to a constant temperature and to daily temperature fluctuations, mainly due to changes in 22:6n-3 and 20:5n-3. The 20:4n-6 levels in oyster gills increased as temperature rose, suggesting an increasing availability of this fatty acid for eicosanoid biosynthesis during stress responses.     

Low temperature acclimation of root fatty acid composition, leaf water potential, gas exchange and growth of soybean seedlings

Abstract Root fatty acid composition, photosynthesis, leaf water potentials, stomatal resistances, leaf specific weights, and root: shoot ratios of soybean were measured in two temperature regimes. Groups of soybean plants were grown in controlled chambers of the Duke University Phytotron under two thermoperiods. One group of the plants was grown from seed for 3 weeks in either 29/23°C or 17/11°C thermoperiods, and another group was grown for 2 weeks in 29/23°C and then transferred to the 17/11°C thermoperiod where it remained for 8 days. Broccoli was also grown in either 29/23°C or 17/11°C thermoperiods. Soybean roots contained more unsaturated fatty acids than broccoli roots, although broccoli roots showed a larger increase in unsaturation than soybean roots with decreased temperature. The fatty acid unsaturation in the roots of soybean began to increase rapidly after the temperature regime was changed. The increase was in the new roots produced in the cold regime rather than in the pre-existing roots. The soybean leaf water and osmotic potentials decreased about 0.4 MPa, beginning one day after the transfer from 29/23°C to 17/11°C, but recovered significantly after 8 d. Plants grown at 17/11 °C had lower rates of photosynthesis and adaxial stomatal resistances, but higher root: shoot ratios and specific leaf weights compared to plants grown at 29/23°C. Plants grown and maintained at 29/23°C showed a steady increase in photosynthetic rates over the 8-d experimental period, whether rates were measured in 1 mol m^?3 or 9 mol m^?3 oxygen. Plants transferred to 17/11°C however maintained constant rates of photosynthesis at 1 mol m^?3 O₂, whereas at 9 mol m^?3 rates declined for 2 d then were constant for the remaining 6 d of the experimental period. These results suggest that changes in membrane fatty acid unsaturation is an important aspect of plant acclimation to chilling temperatures in terms of maintaining root permeability and water uptake. However, the degree of unsaturation is not a good indicator of differences in chilling tolerance among species. The apparent acclimation of photorespiration to a constant percentage of photosynthesis suggests a role of photorespiration in the plant.     

The effect of temperature on hairy root cultures of Catharanthus roseus: Growth,indole alkaloid accumulation and membrane lipid composition

Cultivation of Catharanthus roseus hairy root cultures at different temperatures was found to have an effect on growth rate and indole alkaloid content as well as lipid composition. When lowering the temperature, the roots responded by increasing the degree of unsaturation of cellular lipids, which was mainly due to an increased proportion of linolenic acid in the main lipid classes. The modifications in lipid composition were obviously necessary for the roots to retain the proper cell membrane fluidity at each temperature. Despite of changes in membrane lipids, no effect on the distribution of indole alkaloids between the roots and the medium could be detected. Instead, the level of alkaloid accumulation showed a clear increase with lowering temperature.Abbreviations PC phosphatidylcholine - PE phosphatidylethanolamine - PI phosphatidylinositol - PS phosphatidylserine - PG phosphatidylglycerol - CL cardiolipin - DGD digalactosyldiglyceride - MGD monogalactosyldiglyceride - NL neutral lipids - DU degree of fatty acid unsaturation     

Cold acclimation of Ilex aquifolium under natural conditions with special regard to the photosynthetic apparatus

Frost resistance of leaves of holly ( Ilex aquifolium L.) increased from about −9°C in late summer to −24°C in mid-winter. The gradual rise in cold hardiness occurred when the minimum air temperature dropped to 0°C or below and was closely related to increase in the cellular sap concentration. Predominantly, the decrease in the osmotic potential of the cellular sap was caused by sugar accumulation, mainly of sucrose. The capacity of net photosynthesis of the leaves, as well as the total lipid and protein content and the proportion of individual lipids of the thylakoid membranes, did not significantly change during cold acclimation. The gradual shift towards desaturation in the fatty acids of the thylakoid lipids during the hardening period was neither correlated with alterations in the frost resistance nor did it affect the potential efficiency for various light-induced chloroplast membrane reactions such as linear photosynthetic electron transport, photophosphorylation and the proton gradient (ΔpH). It is suggested that in holly leaves reduction in cell volume changes during freeze-thawing and cryoprotection by sugars could play a dominant role for the increase in frost resistance. Seasonal changes in the degree of unsaturation of polar lipids of the thylakoids could contribute to maintain optimal functional efficiency of the membranes at low temperatures rather than to avoid freezing damage.     

Temperature adaptation of biological membranes. Compensation of the molar activity of cytochrome c oxidase in the mitochondrial energy-transducing membrane during thermal acclimation of the carp (cyprinus carpio L.)

The phospholipid composition, fatty acid pattern and cholesterol content are studied in mitochondria of red lateral muscle of carp acclimated to high and low environmental temperatures.The results of the experiments are: mitochondria from cold-acclimated carp contain higher proportions of ethanolamine phosphatides than mitochondria from warm-acclimated fish, the opposite is true for the choline phosphatides. Thus, at constant pH, the membrane phospholipids are slightly more negatively charged at low acclimation temperature. The total plasmalogen content is reduced in the cold; this reduction is caused by a decrease in the proportion of the choline plasmalogens. The ethanolamine phosphoglycerides contain approx. 20% of the alk-1-enyl acyl type, irrespective of the acclimation temperature. There is no temperature-dependent difference in the low proportion of cholesterol.The fatty acids of total mitochondrial phospholipids are characterized by large amounts of the n-3 and n-6 families. The ratio of unsaturated to saturated fatty acids and the unsaturation index are remarkably higher than those reported for comparable mammalian phospholipids. Cold acclimation of carp does not significantly increase the unsaturation of total phospholipids. A fatty acid analysis of the main isolated phospholipids, however, shows that cold acclimation considerably increases unsaturation of the neutral phosphatidylcholine, whereas it dramatically decreases unsaturation of the negatively charged cardiolipin. It is suggested that the observed fatty acid substitution in phosphatidylcholine indicates a temperature-induced fluidity adaptation within the mitochondrial lipid bilayer, whereas the inverse acclimation pattern of cardiolipin provides a suitable lipid to accommodate the temperature-dependent modifications in the dynamic surface shape of integral membrane proteins.     

Temperature adaptation of biological membranes. The effects of acclimation temperature on the unsaturation of the main neutral and charged phospholipids in mitochondrial membranes of the carp (cyprinus carpio L.)

The phospholipid composition, fatty acid pattern and cholesterol content are studied in mitochondria of red lateral muscle of carp acclimated to high and low environmental temperatures.The results of the experiments are: mitochondria from cold-acclimated carp contain higher proportions of ethanolamine phosphatides than mitochondria from warm-acclimated fish, the opposite is true for the choline phosphatides. Thus, at constant pH, the membrane phospholipids are slightly more negatively charged at low acclimation temperature. The total plasmalogen content is reduced in the cold; this reduction is caused by a decrease in the proportion of the choline plasmalogens. The ethanolamine phosphoglycerides contain approx. 20% of the alk-1-enyl acyl type, irrespective of the acclimation temperature. There is no temperature-dependent difference in the low proportion of cholesterol.The fatty acids of total mitochondrial phospholipids are characterized by large amounts of the $\text{n-3}\text{and}\text{n-6}$ families. The ratio of unsaturated to saturated fatty acids and the unsaturation index are remarkably higher than those reported for comparable mammalian phospholipids. Cold acclimation of carp does not significantly increase the unsaturation of total phospholipids. A fatty acid analysis of the main isolated phospholipids, however, shows that cold acclimation considerably increases unsaturation of the neutral phosphatidylcholine, whereas it dramatically decreases unsaturation of the negatively charged cardiolipin. It is suggested that the observed fatty acid substitution in phosphatidylcholine indicates a temperature-induced fluidity adaptation within the mitochondrial lipid bilayer, whereas the inverse acclimation pattern of cardiolipin provides a suitable lipid to accommodate the temperature-dependent modifications in the dynamic surface shape of integral membrane proteins.     

Lipid response to short-term chilling shock and long-term chill hardening in Jatropha curcas L. seedlings

Low non-freezing temperature is one of the major environmental factors that affect metabolism, growth, development and geographical distribution of chilling-sensitive plants, Jatropha curcas, a chilling-sensitive plant, which is considered as a sustainable energy plant with great potential for biodiesel production. Our previous studies showed that short-term chilling shock at 5 °C for 4 h and long-term chill hardening at 12 °C 1 or 2 days could improve chilling tolerance of J. curcas seedlings, but lipidomic response to chilling shock and chill hardening has not been elucidated. In this study, membrane lipid composition change in J. curcas seedlings during chilling shock and chill hardening was investigated by liquid chromatography-electrospray ionization-mass spectrometry (LC–ESI–MS) approach. The results indicated that the relative abundances of nine classes and 72 species of membrane lipids, such as phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylinositol (PI), lysophosphatidylcholine (lysoPC) and lysophosphatidylglycerol (lysoPG), two glycolipids digalactosyldiacylglycerol (DGDG) and monogalactosyldiacylglycerol (MGDG) and a sulfoquinovosyldiacylglycerol (SQDG), were significantly changed, and the degree of unsaturation of above-mentioned cellular membrane lipids with fatty acid differing in chain lengths and the number of double bonds also increased in varying degrees during chilling shock and chill hardening. These results suggested that remodeling and increase in the degree of unsaturation of membranes lipids may be a common physiological basis for short-term chilling shock- and long-term chill hardening-induced chilling tolerance of J. curcas seedlings.     

The effects of temperature acclimation on membrane sterols and phospholipids of Neurospora crassa

Experiments were conducted to examine the effects of temperature acclimation on sterol and phospholipid biosynthesis in Neurospora crassa. Cultures grown at high (37 degrees C) and low (15 degrees C) temperatures show significant differences in free and total sterol content, sterol/phospholipid ratios and distribution of major phospholipid species in total lipids and two functionally distinct membrane fractions. The ratio of free sterols to phospholipids in total cellular lipids from 15 degrees C cultures was found to be about one-half that found at 37 degrees C, whereas sterol/phospholipid ratios of mitochondrial and microsomal membranes were found to be higher at the low growth temperature. Total sterol and phospholipid biosynthetic rates showed parallel reductions in cultures acclimating to a shift from 37 to 15 degrees C growth conditions. Distribution of [14C]acetate label into free sterols was significantly lower under these conditions, however; indicating an increase in the conversion rate of sterols to sterol esters at the lower temperature. Mitochondrial and microsomal membrane fractions showed distinct phospholipid distributions which also differed from total lipid distributions at the two growth temperatures. In each case there was a consistent decrease in phosphatidylcholine and a corresponding increase in phosphatidylethanolamine as growth temperatures were lowered.     

Seasonal changes in lipid composition and glycogen storage associated with freeze-tolerance of the earthworm, Dendrobaena octaedra

The earthworm, Dendrobaena octaedra, is a common species in the uppermost soil and humus layers of coniferous forests and tundra in temperate and subarctic regions. The species is freeze-tolerant and may survive several months in a frozen state. Upon freezing, glycogen reserves are rapidly converted to glucose serving as a cryoprotectant and fuel for metabolism. In the present study we investigated the induction of freeze-tolerance under field conditions, and sought to find relationships between temperature, glycogen and fat reserves, membrane phospholipid composition and the degree of freeze-tolerance. Freeze-tolerance was induced when worms had experienced temperatures below 5°C for 2 weeks or more. Freeze-tolerance was linked to the magnitude of glycogen reserves, which also fluctuated with field temperatures being highest in autumn and winter. On the other hand fat reserves seemed not to be linked with freeze-tolerance at all. However, high glycogen alone did not confer freeze-tolerance; alterations in the membrane phospholipid fatty acid composition (PLFA) were also necessary in order to secure freeze-tolerance. The changes in PLFA composition were generally similar to changes occurring in other ectothermic animals during winter acclimation with an increased degree of unsaturation of the PLFAs.     

Thermal acclimation,mitochondrial capacities and organ metabolic profiles in a reptile (<Emphasis Type="Italic">Alligator mississippiensis</Emphasis>)

Reptiles thermoregulate behaviourally, but change their preferred temperature and the optimal temperature for performance seasonally. We evaluated whether the digestive and locomotor systems of the alligator show parallel metabolic adjustments during thermal acclimation. To this end, we allowed juvenile alligators to grow under thermal conditions typical of winter and summer, providing them with seasonally appropriate basking opportunities. Although mean body temperatures of alligators in these groups differed by approximately 10°C, their growth and final anatomic status was equivalent. While hepatic mitochondria isolated from cold-acclimated alligators had higher oxidative capacities at 30°C than those from warm-acclimated alligators, the capacities did not differ at 20°C. Cold acclimation decreased maximal oxidative capacities of muscle mitochondria. For mitochondria from both organs and acclimation groups, palmitate increased oligomycin-inhibited respiration. GDP addition reduced palmitate-uncoupled rates more in liver mitochondria from warm- than cold-acclimated alligators. In muscle mitochondria, carboxyatractyloside significantly reduced palmitate-uncoupled rates. This effect was not changed by thermal acclimation. The aerobic capacity of liver, skeletal muscle and duodenum, as estimated by activities of cytochrome c oxidase (COX), increased with cold acclimation. At acclimation temperatures, the activities of COX and citrate synthase (CS) in these organs were equivalent. By measuring COX and CS in isolated mitochondria and tissue extracts, we estimated that cold acclimation did not change the mitochondrial content in liver, but increased that of muscle. The thermal compensation of growth rates and of the aerobic capacity of the locomotor and digestive systems suggests that alligators optimised metabolic processes for the seasonally altered, preferred body temperature. The precision of this compensatory response exceeds that typically shown by aquatic ectotherms whose body temperatures are at the mercy of their habitat.