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Biomedical subjects

J K Raison

Publications and source records attributed to J K Raison.

At least 19 recordsLinked to original sources

Membrane function in mammalian hibernation.

For homeotherms the maintenance of a high, uniform body temperature requires a constant energy supply and food intake. For many small mammals, the loss of heat in winter exceeds energy supply, particularly when food is scarce. To survive, some animals have developed a capacity for adaptive hypothermia in which they lower their body temperature to a new regulatory set-point, usually a few degrees above the ambient. This process, generally known as hibernation, reduces the temperature differential, metabolic activity, as well as the energy demand, and thus facilitates survival during winter. Successful hibernation in mammals requires that the enzymatic processes are regulated in such a manner that metabolic balance is maintained at both the high body temperature of the summer-active animal (37 degrees C) and the low body temperature of the winter-torpid animal (approx. 5 degrees C). This means that the cellular membranes have thermal properties capable of maintaining a balanced metabolism at these extreme physiological temperatures. The available evidence indicates that, for some tissues, preparation for hibernation involves an alteration in the lipid composition and thermal properties of cellular membranes. Marked differences in the thermal response of cellular membranes have been observed on a seasonal basis and, in some membranes, differences in lipid composition have been associated with the torpid state. However, to date, no consistent changes in lipid composition which would account for, or explain, the changes in membrane thermal response, have been detected. An important point to emphasize is that the process of 'homeoviscous adaptation', which occurs in procaryotes and some poikilotherms during acclimation to low temperatures, is not a characteristic feature of most membranes of mammalian hibernators.

Animals↗

Sensitivity of Altitudinal Ecotypes of the Wild Tomato Lycopersicon hirsutum to Chilling Injury.

The transition temperature of the leaf polar lipids and the critical temperature for chill-induced inhibition of photosynthesis was determined for three altitudinal ecotypes of the wild tomato Lycopersicon hirsutum. Photosynthesis was measured as CO(2)-dependent O(2) evolution at 25 degrees C after leaf slices were exposed to chilling temperatures for 2 hours at a moderate photon flux density of 450 micromoles per square meter per second. The transition temperature of the leaf polar lipids was detected from the change in the temperature coefficient of the fluorescence intensity of trans-parinaric acid. Chill-induced photoinhibition was evident in the three tomato ecotypes when they were chilled below a critical temperature of 10 degrees , 11 degrees , and 13 degrees C, respectively, for the high (LA1777), mid (LA1625), and low (LA1361) altitudinal ecotypes. The temperature differential, below the critical temperature, required to produce a 50% inhibition was also similar for the three ecotypes. A transition was detected in the leaf polar lipids of these plants at a temperature similar to that of the critical temperature for photoinhibition. The results show that the three tomato ecotypes are similar with respect to their critical temperature for chilling-induced photoinhibition and the rate of their response to the chilling stress. They are, thus, similarly sensitive to chilling.

Journal Article↗

The influence of storage temperature on the transition, activation enthalpy, and activity of enzymes associated with inner mitochondrial membranes.

The effects of storage at low temperature on the transition in enzyme function, Tf*, and the Arrhenius activation energy, Ea, were determined for several enzymes associated with the inner membrane of rat liver mitochondria. The enzymes studied were succinate:cytochrome c reductase, cytochrome c oxidase, beta-hydroxybutyrate dehydrogenase, and oligomycin-sensitive, Mg2+-activated ATPase. For freshly isolated mitochondria the Tf*, for succinate:cytochrome c reductase and cytochrome c oxidase, occurred at approximately 23 degrees C and was coincident with a transition in structure, Ts*, determined as the change in temperature coefficient of motion for a spin label intercalated with the membrane lipids. This suggest that the change in thermal response of the membrane-associated enzymes is related to a change in molecular ordering of the membrane lipids. When mitochondria were stored at -12 degrees C, the specific activities of succinate:cytochrome c reductase and cytochrome c oxidase decreased. Concomitant with these changes the Ea, above Tf*, increased. After 100 days storage at -12 degrees C, Ea above Tf* approached the value for Ea below Tf* such that the transition in thermal response could no longer be detected. In contrast, for mitochondria stored at -196 degrees C, although the specific activity declined over the 100 days storage, no changes in either Ea or Tf* were evident. The results indicate a need for caution in evaluating comparative studies of Tf and Ea, for membrane-associated enzymes, using mitochondria which have been frozen and stored.

Animals↗

Mitochondrial membrane transitions in heart and other organs of a hibernator.

Critical temperatures (T) for transitions in both lipid structure and enzyme function of mitochondrial membranes from liver, kidney, brown fat, and heart tissues were determined for the hibernator Spermophilus lateralis at two weekly intervals from early summer to late autumn and during hibernation. For all tissues T fell into one of three groups: those below 4 degrees C (the minimal level of accurate determination), those centered about a mean of 11.9 +/- 1.4 degrees C, and those centered about a mean of 20.9 +/- 1.8 degrees C. The T for tissues from torpid animals and from heart, at all sampling periods, was below 4 degrees C. For liver, kidney, and brown fat the mean T was approximately 21 degrees C in early summer but was lowered later in the season in a two-step process, falling to below 4 degrees C before the animals were exposed to cold and entering torpor. It is concluded that for mitochondria the thermal response of the membrane lipids is altered such that the transition in structure and function is always below the minimum body temperature likely to be experienced by this animal. Heart tissue is exceptional in that the transition is at a temperature consistent with a body temperature of torpor even in summer-active animals.

Animals↗

Radiation-induced lipid peroxidation and the fluidity of erythrocyte membrane lipids.

The effect of radiation-induced peroxidation on the fluidity of the phospholipids of the erythrocyte membrane was studied using both erythrocyte ghosts and liposomes formed from the polar lipids of erythrocytes. In liposomes, the oxidation of the phospholipids increased with radiation dose, but there was no change in the fluidity of the lipids as measured by spin-label motion. Under the same conditions of irradiation, no oxidation of phospholipid was detected in erythrocyte ghosts, although changes occurred in the motion of spin labels intercalated with the membrane. These changes were attributed to radiation-induced alterations in the membrane proteins. It is concluded that alterations in motion of spin labels, observed with intact membranes after irradiation, are most likely the result of changes in the structure of membrane proteins rather than the lipids.

Dose-Response Relationship, Radiation↗

Compositional and Thermal Properties of Thylakoid Polar Lipids of Nerium oleander L. in Relation to Chilling Sensitivity.

The polar lipid classes from thylakoids of Nerium oleander L. were studied with the aim of relating changes in their composition and thermal behavior with reported changes in the transition temperature of their polar lipids and chilling sensitivity of their leaves. With an increase in growth temperature, the transition temperature of phosphatidylglycerol increased from 16 degrees C to 26 degrees C, and for sulfoquinovosyldiacylglycerol from 19 degrees C to 24 degrees C. Transitions in the other lipid classes were below -10 degrees C for plants grown at both growth temperature. The major changes in the molecular species of phosphatidylglycerol, with increasing growth temperature, were an increase in 1-oleoyl-2-palmitoyl phosphatidylglycerol from 21 to 39% and a decrease in 1-oleoyl-2-trans-3-hexadecanoic phosphatidylglycerol from 51 to 25%. Although the disaturated species increased from 8 to 23%, the maximum was less than that reported for chilling-sensitive plants. There was no change in the sum of the palmitic, hexadeca-trans-3-enoic and stearic acids. Dipalmitoyl sulfoquinovosyldiacylglycerol increased from 12 to 20% and 1-linolenoyl-2-palmitoyl sulfoquinovosyldiacylglycerol decreased from 40 to 30%. It is concluded that the increase in the transition temperature of the polar lipids and the sensitivity of acclimated oleander plants to chilling could not be predicted by the absolute sum of the saturated fatty acids or disaturated molecular species in phosphatidylglycerol. The polar lipid transition appears to be a product of mixing of both high and low melting-point lipids.

Journal Article↗

Phase transitions in thylakoid polar lipids of chilling-sensitive plants: a comparison of detection methods.

The phase behavior of thylakoid polar lipids from plants sensitive to chilling injury was investigated by calorimetry, electron spin resonance spectroscopy of spin labels, and fluorescence intensity after labeling with trans-parinaric acid. The plants used were oleander (Nerium oleander), mung bean (Vigna radiata L. var Mungo), and tomato (Lycopersicon esculentum cv Grosse Lisse). For all plants the initiation temperature for the calorimetric exotherm was coincident (+/-1 degrees C) with the transition determined by the increase in the temperature coefficient of spin label motion and fluorescence intensity of trans-parinaric acid. For oleander plants, grown at 45 degrees C, the transition was at 7 degrees C while for plants from the same clone, grown at 20 degrees C, it was at -2 degrees C. For mung bean and tomato the transition was between 9 and 12 degrees C. The similarity in the transition detected by spin labeling and fluorescence intensity suggest that spin labels, like the fluorescent label trans-parinaric acid, preferentially partition into domains of ordered lipid. The coincidence of the temperature for initiation of the transition, determined by the three techniques, shows that each is a valid method of assessing a phase transition in membrane polar lipids.

Journal Article↗

Phase transitions in liposomes formed from the polar lipids of mitochondria from chilling-sensitive plants.

The thermal response of mitochondrial polar lipids from a variety of chilling-sensitive and chilling-insensitive plants was determined by differential scanning calorimetry. A phase transition was observed at 15 degrees C for mitochondria from soybeam (Glycine max. cv Davis) hypocotyl, at 16 degrees C for tomato (Lycopersicon esculentum cv Flora-Dade and cv Grosse Lisse) fruit, at 15 degrees C for cucumber (Cucumus sativus L.) fruit, at 14 degrees C for mung bean (Vigna radiata var Berken) hypocotyl, and at 15 degrees C for sweet potato (Ipomea batatas L.) roots. The transition temperature was not significantly altered by the scan rate and was reversible. Changes in the temperature coefficient of motion for a spin label, intercalated with the polar lipids, occurred at a temperature slightly below that of the phase transition, indicating that the polar lipids phase separate below the transition. No phase transition was observed for mitochondrial polar lipids from barley (Hordeum vulgare) roots, wheat (Triticum aestivum L. cv Falcon) roots, and Jerusalem artichoke (Helianthus tuberosus L.) tubers. The results show that a phase change occurs in the membrane lipids of mitochondria a few degrees above the temperature below which chilling injury is evident in the sensitive species. Thus they are consistent with the hypothesis that sensitivity to chilling injury is related to a temperature-induced alteration in the structure of cell membranes.

Journal Article↗

Thermal behavior and lipid composition of cauliflower plasma membranes in relation to ATPase activity and chilling sensitivity.

A plasma-membrane fraction rich in ion-stimulated ATPase activity was isolated from cauliflower (Brassica oleracea L.) buds. The activity of the ATPase was dependent on Mg(2+) and stimulated 4-fold by K(+). The lipids of the membrane fraction contained 57% by weight of phospholipid, 16% glycolipid including sterol glycosides, and 27% neutral lipids. Sterols and sterol esters comprised 9% by weight of the total lipid fraction, and the m ratio of total sterol to phospholipid was 0.5. Fatty acid unsaturation of the membrane lipids was 75%. Arrhenius plots of the Mg(2+) and Mg(2+) + K(+) stimulated ATPase activity were biphasic with an increase in activation energy occurring below about 12 degrees C, a response typical of some membrane-associated enzymes of chilling-sensitive plants. No thermal transitions were detected in the membranes or membrane lipids between 0 and 30 degrees C using differential scanning calorimetry and electron spin resonance spectroscopy. This type of thermal behavior is typical of membranes of chilling-resistant plants. It was concluded that the low temperature increase in activation energy of the ion-stimulated, membrane-associated ATPase is an intrinsic property of the enzyme system and not the result of a transition in the bulk membrane lipid.

Journal Article↗

Growth Temperature-Induced Alterations in the Thermotropic Properties of Nerium oleander Membrane Lipids.

The temperature boundary for phase separation of membrane lipids extracted from Nerium oleander leaves was determined by analysis of spin label motion using electron spin resonance spectroscopy and by analysis of polarization of fluorescence from the probe, trans-parinaric acid. A discontinuity of the temperature coefficient for spin label motion, and for trans-parinaric acid fluorescence was detected at 7 degrees C and -3 degrees C with membrane lipids from plants grown at 45 degrees C/32 degrees C (day/night) and 20 degrees C/15 degrees C, respectively. This change was associated with a sharp increase in the polarization of fluorescence from trans-parinaric acid indicating that significant domains of solid lipid form below 7 degrees C or -3 degrees C in these preparations but not above these temperatures. In addition, spin label motion indicated that the lipids of plants grown at low temperatures are more fluid than those of plants grown at higher temperatures.A change in the molecular ordering of lipids was also detected by analysis of the separation of the hyperfine extrema of electron spin resonance spectra. This occurred at 2 degrees C and 33 degrees C with lipids from the high and low temperature grown plants, respectively. According to previous interpretation of spin label data the change at 29 degrees C (or 33 degrees C) would have indicated the temperature for the initiation of the phase separation process, and the change at 7 degrees C (or -3 degrees C) its completion. Because of the present results, however, this interpretation needs to be modified.Differences in the physical properties of membrane lipids of plants grown at the hot or cool temperatures correlate with differences in the physiological characteristics of plants and with changes in the fatty acid composition of the corresponding membrane lipids. Environmentally induced modification of membrane lipids could thus account, in part, for the apparently beneficial adjustments of physiological properties of this plant when grown in these regimes.

Journal Article↗

Maintenance of Membrane Fluidity during Development of Freezing Tolerance of Winter Wheat Seedlings.

Fluidity of membrane lipids of shoot and root tissue and of chloroplasts from young wheat seedlings of contrasting freezing tolerance was investigated by measuring the motion and order parameters after spin labeling. A striking similarity was observed in membrane lipid fluidity of the five cultivars grown at 22 C. After cold hardening by growth at 2 C, a small change in membrane lipid fluidity was observed, but this was not correlated with the development of freezing tolerance, and there was no alteration in the transition temperature of membrane lipids. The results show that neither changes in membrane lipid fluidity nor transition temperature are a necessary feature of cold acclimation in wheat.

Journal Article↗

Correlation between Changes in Mitochondrial Membranes of Artichoke Tubers and Their Hardening and Dormancy.

Spin labeling studies using mitochondrial membranes of Jerusalem artichoke (Helianthus tuberosus L.) showed that the decrease during winter in the temperatures of the upper and lower lipid transitions correlated with the development of freezing hardiness of the tubers. The killing temperature for tuber tissue reached a minimum of -12 C, about 5 C degrees lower than the lower transition. Freeze-hardiness decreased when the lower transition increased at the time of sprouting.Low temperature storage was not required to induce freeze-hardiness or to lower the transitions. These changes occurred in tubers under field conditions and at a constant growth temperature of 25 +/- 1 C. In both conditions, tuber dormancy preceded the mitochondrial membrane changes.In young nondormant tubers, cessation of growth by storage at 0 or 4 C and the induction of dormancy with abscisic acid led to decreased temperatures of the upper and lower transitions.Changes in Arrhenius activation energy of succinate oxidase were correlated with the seasonal changes in the upper and lower transitions confirming that these were indeed a reflection of altered thermal responses of mitochondrial membranes, which might be part of a general mechanism in this plant of insuring membrane integrity during the freeze-thaw process. In contrast with chilling injury, there is no precise correlation between the temperature of the lower transition and the temperature below which freezing injury occurs.

Journal Article↗

The fluidity of chloroplast thylakoid membranes and their constituent lipids: a comparative study by ESR.

Comparative measurements were made of the fluidity of chloroplast thylakoids, total membrane lipids and polar lipids utilizing the order parameter and motion of spin labels. No significant differences were found in the fluidity of membranes or total membrane lipids from a wild type and a mutant barley (Hordeum vulgare chlorina f2 mutant) which lacks chlorophyll b and a 25 000 dalton thylakoid polypeptide. Redistribution of intrinsic, exoplasmic face (EF) membrane particles by unstacking thylakoid membranes in low salt medium also had no effect on membrane fluidity. However, heating of isolated thylakoids decreased membrane fluidity. The fluidity of vesicles composed of membrane lipids is much greater than that of the corresponding membranes. Fluidity of the membranes, however, increased during greening indicating that the rigidity of the membranes, compared with that of total membrane lipids, is not caused by chlorophyll or its associated peptides. It is concluded that the restriction of motion in the acyl chains in the thylakoids is not caused by chlorophyll or the major intrinsic polypeptide but by some other protein components.

Chloroplasts↗

Membrane lipid fluidity and its effect on the activation energy of membrane-associated enzymes.

1. The fatty acid composition of mitochondrial membranes from sheep and rats was altered by feeding these animals diets which were rich in unsaturated fatty acids. Changes in membrane lipid fluidity resulting from the altered membrane lipid composition were assessed by determining the upper temperature limit of the disorder-order transition (Tf) and the Arrhenius activation energy (Ea) of succinate oxidase. 2. After feeding the unsaturated fatty acid-rich diet to sheep the Ea, in the temperature range above Tf, increased from 8 to 63 kJ . mol-1 while Tf decreased from 32 to 15 degrees C. Rats fed an unsaturated fatty acid-rich diet exhibited an increase in Ea from 17 to 63 kJ . mol-1 and a decrease in Tf from 23 to 4 degrees C. 3. This decrease in Tf was related to an increase in the ratio of linoleic acid to stearic acid in the membrane lipid. Tf was not related to the proportion of unsaturated fatty acids in the membrane lipids, although an increase in unsaturation usually led to a decrease in Tf. 4. The results show that membrane lipid fluidity has a direct influence on the conformation of the active site of some membrane-associated enzymes, with the result that such enzymes display a higher Ea when the membrane lipids are comparatively more fluid. The increase in Ea of membrane-associated enzymes which accompanies changes in the physical state of membrane suggests that some proteins may phase separate with the more fluid lipids at low temperatures.

Animals↗

Seasonal Changes in the Structure and Function of Mitochondrial Membranes of Artichoke Tubers: A Requisite for Surviving Low Temperatures during Dormancy.

The temperature limits of the order-disorder transition, and the Arrhenius activation energy of succinate oxidase activity for mitochondria of Jerusalem artichoke (Helianthus tuberosus L.) tubers were determined from the initiation to the termination of dormancy. The temperature limits for the transition at the initiation of dormancy were 25 and 3 C. These changed to 9 and -5 C at mid-dormancy and returned to 25 and 2 C at the termination of dormancy. The Arrhenius activation energy measured in the temperature range above the transition was 35 kilojoules per mole at middormancy and decreased to 17 kilojoules per mole at the termination of dormancy when sprouting was evident. The coincidence of the changes in membrane structure and function with dormancy suggests that artichokes possess a mechanism for regulating membrane lipid structure so that cellular integrity of tuber tissue is maintained even when the tubers are exposed to low temperatures.

Journal Article↗

Seasonal changes in the structure and function of mitochondrial membranes of artichoke tubers: acyl Fatty Acid composition and the effect of growth conditions.

Changes in the temperature response, fluidity, function and the acyl fatty acid composition, were determined for a mitochondria-rich membrane fraction from Jerusalem artichoke (Helianthus tuberosus L.) tubers during dormancy for a crop which matured in midsummer. The temperature of both the upper and lower limits of the membrane lipid transition decreased during dormancy from 26 C and 1 C to 4 C and -5 C, respectively. This was similar to the changes observed with crops maturing in late autumn. The order parameter of a spin label intercalated into the membrane lipids decreased from about 0.6 to 0.5 during dormancy and returned to the original value before sprouting, showing that membrane fluidity increased during dormancy. The activation energy of succinate oxidase of tuber mitochondria was generally high at middormancy when membrane lipids were more fluid and decreased as the membranes became more rigid at the end of dormancy. The fatty acid composition of the membrane lipids did not alter significantly during dormancy. The results indicate that neither decreasing day length nor low soil temperature during tuber maturation is essential for the initiation of the membrane changes necessary for tubers to avoid low temperature injury during dormancy. The increase in membrane fluidity during dormancy could not be accounted for by an increase in the proportion of unsaturated fatty acids in the membrane lipids.

Journal Article↗

Seasonal changes in membrane lipid transitions and thyroid function in the hedgehog.

Upper (Tf) and lower (Ts) temperature limits of order-disorder transitions in blood cell lipids of hedgehogs, Erinaceus europaeus, were determined over an annual cycle. There was a significant decrease in the temperature of both Tf and Ts from values of 19 and 6 degrees C, respectively, for summer animals to values of 14 and -2.0 degrees C for winter animals. Plasma thyroxine levels decreased from a summer mean of 16.0 nmol/liter to a mean of 2.3 nmol/liter in winter. Basal oxygen consumption also decreased from the summer mean 0.45 ml/g body wt/h to a mean 0.39 ml/g body wt/h in winter. In winter a group of hedgehogs kept indoors at room temperature was compared with a group kept outdoors exposed to natural winter conditions, and there was no significant difference between them in the above parameters. We conclude that the winter membrane lipid and metabolic changes are not a response to low temperature per se but part of a circannual homeostatic adjustment at least partly regulated by thyroid hormone.

Animals↗