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Rapid changes in the phospholipid composition of gill membranes during thermal acclimation of the rainbow trout, Salmo gairdneri.

The phospholipid composition of gill tissue was determined in rainbow trout (Salmo gairdneri) undergoing thermal acclimation between 5 degrees C and 20 degrees C for a period of up to 28 days. Proportions of phosphatidylethanolamine (PE) and cardiolipin (CL) increased during cold acclimation and decreased during warm acclimation; proportions of phosphatidylcholine (PC) changed in the opposite direction (i.e., decreased during cold acclimation). In contrast, levels of phosphatidylserine, -inositol, and sphingomyelin did not vary significantly. Thermal modulation of headgroup composition occurred rapidly as reflected by changes in the ratio of PC-to-PE, which rose significantly from 2.40 +/- 0.09 to 2.92 +/- 0.09 within 72 h of transfer from 5 to 20 degrees C; adaptation to 5 degrees C was equally rapid. Proportions of PE changed more rapidly than those of PC during cold adaptation, whereas the opposite was true during warm acclimation. Both the time course and the direction of the observed changes in phospholipid composition suggest that such adjustments may contribute to the homeoviscous regulation of membrane properties, particularly during the initial stages of thermal adaptation.

Acclimatization↗

The effects of temperature- and oxygen-acclimation on phospholipids of goldfish (Carassius auratus L.) brain microsomes.

Brain microsome phospholipids and their acyl groups, from temperature and oxygen acclimated goldfish, were investigated. At the lower acclimation temperature (5C) the proportion of ethanolamine- to choline-glycerophosphatides (GPE/GPC) was increased, and the proportion of phosphatidal ethanolamine value decreased. A rise in the n-6/n-3 fatty acyl group also occurred in cold acclimation. Irrespective of acclimation temperature, 25 degrees C or 5 degrees C, a partial replacement of GPC by GPE occurred when the concentration of oxygen was increased; conversely the GPE/GPC ratio decreased at the hypoxic level. The plasmalogen GPE content increased as the oxygen concentration was raised. A rise in the n-6/n-3 ratio, for ethanolamine glycerophosphatides and phosphatidyl choline, occurred when the oxygen concentration was increased (hypoxia to hyperoxia). It is concluded that the lipid alterations associated with thermal acclimation are, in part, attributable to the concomitant change in oxygen concentration.

Acclimatization↗

Cold-acclimation-induced protein hypertrophy in channel catfish and green sunfish.

1. Following acclimation of channel catfish to a reduction in temperature from 25 degrees to 15 degrees C, there were approximately two-fold increases in liver mass, cell size, total protein, and total enzyme activity, relative to activity per milligram of protein and per gram wet weight of tissue, indicating tissue hypertrophy. There was no change in either total liver DNA content or protein concentration per gram weight. 2. Green sunfish, unlike catfish, showed virtually no change in liver mass following cold acclimation. However, sunfish showed a net increase in total liver protein content and an increase in protein concentration. The increase in protein content was balanced by a reciprocal and equivalent decrease in glycogen content. Consequently, liver mass was maintained. 3. During cold acclimation both catfish and sunfish showed an increase in ventricular heart mass and protein content, but no change in protein concentration. 4. The activities of several enzymes were measured in liver from 15 degrees C and 25 degrees C steady-state-acclimated catfish and at intervals following transfer from 15 degrees to 25 degrees C and from 25 degrees to 15 degrees C. Total tissue enzyme activity showed positive compensation which correlated with the change in liver mass and protein content. Specific activities based on protein and on wet weight showed dissimilar acclimatory patterns. Two enzymes - cytochrome oxidase and lactate dehydrogenase - showed inverse compensation in specific activity but positive compensation in total activity. Citrate synthase, glucose-6-phosphate-dehydrogenase and 6-phosphogluconate dehydrogenase showed positive compensation in both specific and total activities. 5. The increase in tissue protein content or 'protein hypertrophy' occurred with cell hypertrophy in cold-acclimated catfish, while protein hypertrophy occurred as an increased protein concentration without cell hypertrophy in sunfish. This phenomenon is considered adaptive in that it permits a compensatory increase in the total enzymatic capacity of a tissue. The two-fold increases in total enzyme activities, superimposed on either an increase or decrease in specific activity, suggest that two biochemical mechanisms may be operative during cold-induced liver hypertrophy, one effecting a specific step in protein translation at a point common to the synthesis of all proteins and a second targetted pretranslationally, i.e., transcriptional regulation.

Acclimatization↗

Accelerated Biodegradation of High and Low Concentrations of p-Nitrophenol (PNP) by Bacterial Inoculation in Industrial Wastewater: The Role of Inoculum Size on Acclimation Period

The effect of inoculum size on the acclimation period and rate and extent of p-nitrophenol (PNP) degradation at high (1-10 mg/L) and low (26 &mgr;g/L) concentrations for two bacteria was determined in defined media as well as industrial wastewater. Increased inoculum size did not affect the acclimation period of either bacterium at high (1-10 mg/L) PNP concentrations. At low PNP concentrations (26 &mgr;g/L), the two bacteria behaved differently. The acclimation period was shortened and both the rate and extent of mineralization of PNP were enhanced by increasing the Corynebacterium sp. inoculum size from 3 x 10(5) to 3 x 10(6) cells/ml. Addition of phosphate or elimination of predators also reduced the acclimation period. Conversely, increasing the inoculum size from 3 x 10(5) to 5 x 10(6) cells/ml of Pseudomonas putida lengthened the acclimation period and reduced both the rate and extent of mineralization. It is suggested that, in a given environment, the success of an introduced species to enhance the degradation of a chemical depends upon (i) concentration of the chemical, (ii) selection of an appropriate microorganism, and (iii) utilization of a suitable inoculum size.

Journal Article↗

Plasticity of myosin heavy chain expression with temperature acclimation is gradually acquired during ontogeny in the common carp (Cyprinus carpio L.).

Common carp (Cyprinus carpio L.) were reared from hatching until 61 mm total length (TL) at 21 degrees C. At 14 weeks and 20 weeks post-hatch, corresponding to initial lengths of 30 mm and 44 mm respectively, fish were acclimated to 10 degrees C using a rate of cooling of 1 degrees C per day. A statistical model was used to compare the time course in the change of white muscle myofibrillar ATPase activity with temperature acclimation. The myosin heavy chain (MHC) composition of white muscle myofibrils was investigated using peptide mapping. A significant increase in myofibrillar ATPase activity was observed after 2-3 weeks in the 44 mm group, but not until 4-5 weeks in the 30 mm group. when they had reached 37 mm TL. The MHC banding pattern of 120 mm TL fish acclimated to 10 degrees C or 21 degrees C for a minimum of 6 weeks were distinct from each other. The MHC peptide map characteristic of 10-degrees C-acclimated fish was not observed in individuals less than 37 mm length. We therefore conclude that the capacity to alter the composition and properties of myofibrils with cold acclimation is acquired in juvenile carp at around 37 mm TL.

Acclimatization↗

Alterations of calf venous and arterial compliance following acclimation to heat administered at a fixed daily time in humans.

We investigated the effects of heat acclimation on venous and arterial compliance in humans. Four male and four female volunteers were exposed to an ambient temperature of 40 degrees C and relative humidity of 40% for 4 h (1330-1730 hours) per day for 9-10 consecutive days. The calf venous compliance (CV) was estimated using venous occlusion plethysmography with a mercury-in-silastic strain gauge placed around the right calf at its maximum girth. The compliance of the small (CSA) and large (CLA) arteries were assessed by reflective and capacitance compliance by analyzing the radial artery blood pressure waveforms, basing on the use of a modified Windkessel model. The calf CV, CSA, CLA, systolic and diastolic blood pressures, heart rate and core temperature were determined twice a day, 0930-1100 hours (AM test) and 1500-1630 hours (PM test), in both heat-acclimated and non-heat-acclimated (control) conditions. Heat acclimation appeared to decrease blood pressures, heart rate and significantly lowered core temperature only in the PM test. In the control condition, the calf CV was not affected by the time of day and the CSA was significantly depressed in the PM test. After acclimation to heat, the calf CV significantly increased and the CSA did not decrease in the PM test. The results presented suggest that repeated heat exposure in humans, for 4 h at a fixed time daily, increases the calf CV and the CSA particularly during the period when the subjects were previously exposed to heat.

Adult↗

Monitoring expression profiles of Arabidopsis genes during cold acclimation and deacclimation using DNA microarrays.

A comparative analysis of gene expression profiles during cold acclimation and deacclimation is necessary to elucidate the molecular mechanisms of cold stress responses in higher plants. We analyzed gene expression profiles in the process of cold acclimation and deacclimation (recovery from cold stress) using two microarray systems, the 7K RAFL cDNA microarray and the Agilent 22K oligonucleotide array. By both microarray analyses, we identified 292 genes up-regulated and 320 genes down-regulated during deacclimation, and 445 cold up-regulated genes and 341 cold down-regulated genes during cold acclimation. Many genes up-regulated during deacclimation were found to be down-regulated during cold acclimation, and vice versa. The genes up-regulated during deacclimation were classified into (1) regulatory proteins involved in further regulation of signal transduction and gene expression and (2) functional proteins involved in the recovery process from cold-stress-induced damages and plant growth. We also applied expression profiling studies to identify the key genes involved in the biosynthesis of carbohydrates and amino acids that are known to play important roles in cold acclimation. We compared genes that are regulated during deacclimation with those regulated during rehydration after dehydration to discuss the similarity and difference of each recovery process.

Acclimatization↗

Long-term temperature acclimation of photosynthesis in steady-state cultures of the polar diatom Fragilariopsis cylindrus.

Cultures of the obligate psychrophilic diatom Fragilariopsis cylindrus (Grunow) were grown for 4 months under steady-state conditions at -1 degrees C and +7 degrees C (50 micromol photons m(-2) s(-1)) prior to measurements in order to investigate long-term acclimation of photosynthesis to both temperatures. No differences in maximum intrinsic quantum yield of PS II (F(V)/F(M)) and relative electron transport rates could be detected at either temperature after 4 months of acclimation. Measurements of photosynthesis (relative electron transport rates) vs. irradiance (P vs. E curves) revealed similar values for relative light utilization efficiency (alpha = 0.57 at -1 degrees C, alpha = 0.60 at +7 degrees C) but higher values for irradiance levels at which photosynthesis saturates (E(K)) at -1 degrees C and, therefore, higher maximum photosynthesis (P(MAX) = 54 (relative units) at -1 degrees C, P(MAX) = 49 at +7 degrees C). Nonphotochemical quenching (NPQ) measurements at 385 mumol photons m(-2) s(-1) indicated higher (37%) NPQ for diatoms grown at -1 degrees C compared to +7 degrees C, which was possibly related to a 2-fold increase in the concentration of the pigment diatoxanthin and a 9-fold up-regulation of a gene encoding a fucoxanthin chlorophyll a,c-binding protein. Expression of the D1 protein encoding gene psbA was ca. 1.5-fold up-regulated at -1 degrees C, whereas expression levels of other genes from Photosystem II (psbC, psbU, psbO), as well as rbcL, the gene encoding the Rubisco large subunit were similar at both temperatures. However, a 2-fold up-regulation of a plastid glyceraldehyde-P dehydrogenase at -1 degrees C indicated enhanced Calvin cycle activity. This study revealed for the first time that a polar diatom could efficiently acclimate photosynthesis over a wide range of polar temperatures given enough time. Acclimation of photosynthesis at -1 degrees C was probably regulated similarly to high light acclimation.

Acclimatization↗

Rapid cold-induced changes of membrane order and delta 9-desaturase activity in endoplasmic reticulum of carp liver: a time-course study of thermal acclimation.

The membrane order of liver endoplasmic reticulum (ER) membranes of 10 degrees C- and 30 degrees C-acclimated carp has been compared using the fluorescence polarization technique with DPH as probe. Membranes from cold-acclimated fish displayed lower polarizations than corresponding membranes from warm-acclimated fish, the difference compensating for 34-50% of the direct effects of temperature upon polarization. The changes in delta 9-desaturase activity and fluorescence polarization of DPH in ER membranes have been monitored as a function of time during cold acclimation of 30 degrees C-acclimated carp. Cooling was achieved in three stages over 48 h. Desaturase activity in both rough and smooth ER showed a rapid increase in activity for the first three days followed by a decline on day 4 and a second increase up to day 10. Polarization of DPH (measured at 10 degrees C) was rapidly reduced on cooling with no further change after day 4. The halftime for change in polarization and for the first desaturase induction were both approx. 2 days although large changes in polarization were evident within 24 h after the onset of cooling. During the cooling phases the daily changes in DPH polarization were quantitatively related to increments in desaturase capacity. The second desaturase induction had no effect upon membrane structure, at least as indicated by the polarization technique.

Animals↗

Lipid adaptation in liver mitochondrial membranes of carp acclimated to different environmental temperatures: phospholipid composition, fatty acid pattern and cholesterol content.

The lipid fraction of liver mitochondria has been studied in carp acclimated to high and low environmental temperatures. Evidence is provided for a temperature-induced lipid adaptation which might control membrane fluidity. This supports suggestions made in a recent communication on temperature-induced changes in the Arrhenius functions of mitochondrial oxidase systems from carp liver ((Wodtke, E. (1976) J. Comp. Physiol. 110, 145--157). The results of the analysis of lipid composition are: the ratio of phospholipid to protein does not differ at cold and warm acclimation temperatures. Fish kept at low environmental temperature show a decreased molar ratio of cholesterol : phospholipid in mitochondrial membrane lipids; the diminished complexation renders phospholipids more sensitive to fluidity control by fatty acid substitution. A decrease of mitochondrial phosphatidylcholine at low acclimation temperature is observed, which is compensated by increased amounts of phosphatidylethanolamine and phosphatidylinositol. This means there is an increase in the acidic character of the phospholipids at low environmental temperature, and might be the cause of fluidization of the membrane and a decrease in transition temperature. The fatty acid pattern of carp mitochondria differs markedly from that of mammalian mitochondria; it is not identical for total lipids, phosphatidylcholine and phosphatidylethanolamine, and is roughly characterized by high amounts in n--3, but low amounts in n--6 and mono-unsaturated fatty acids. Low environmental temperature decreases the proportion of saturated species and markedly lowers the ratio of the alpha-linolenic acid family, the latter perhaps being caused by the increased activity of delta6-desaturase, as calculated for low acclimation temperature. An increase in mean unsaturation and in the proportion of 1,2-diunsaturated phospholipids most probably increases membrane fluidity at low acclimation temperature.

Adaptation, Physiological↗

Effect of acclimation temperature on the axon and fiber diameter spectra and thickness of myelin of fibers of the optic nerve of goldfish.

The optic nerves of common goldfish acclimated to 5 and 25 degrees C were fixed with glutaraldehyde in either phosphate buffer or PIPES with EGTA, post-fixed with osmium tetroxide, and examined by electron microscopy. The axon diameter spectra, from axons measured in electron micrographs and those measured on the electron microscope screen, differ noticeably with acclimation temperature. At the lower temperature, there is a definite shift toward the occurrence of larger fibers compared with the spectrum of the 25 degrees C fish. Although the number of fibers assessed is small compared with the total number in the goldfish nerve, these results confirm our previous study. These findings could be attributed to an increase in the number of new fibers during the acclimation to the higher temperature. We discuss this possibility and on the available evidence find it unlikely. Other changes in the axon and fiber are also seen with acclimation temperature. The axon to fiber diameter ratio, made directly from the electron micrographs, shows that axons from the nerves of the higher acclimation temperature fish possess consistently thicker myelin sheaths than are found for axons in nerves of the lower temperature fish. This finding is also in agreement with results obtained by us from measurements independent of each other.

Acclimatization↗

Absence of metabolic acclimation to cycling temperature conditions in the goldfish.

Studies of whole organism metabolism have failed to confirm an effect of cyclic acclimation in fish while studies at the hematological level show clear changes due to cyclic thermal regimes. To reevaluate the effect of cycling temperatures on fish metabolism, oxygen consumption was measured in goldfish acclimated to either a constant temperature (28 degrees C) or a diurnal sinewave cycle (20-36 degrees C). Fish from both acclimation conditions were tested at one of three temperatures (20, 28, 36 degrees C). Oxygen consumption increased with temperature but did so independently of the acclimation regime. Thus, cyclic acclimation did not alter metabolic sensitivity.

Acclimatization↗

Effect of acclimation temperature on oxygen transport in the blood of the carp, Cyprinus carpio.

The temperature dependence of the O2 equilibrium in whole blood was measured in carp acclimated for more than 3 weeks at 10 degrees C or 20 degrees C water temperature. O2 combining curves were obtained for the same samples at 10 degrees C or 20 degrees C using blood from the 10 degrees C or 20 degrees C acclimated fish (group A) or only at the acclimation temperature (group B). Whereas in group B the P50 was about the same at both temperatures, in group A P50 was higher at 20 degrees C than at 10 degrees C, yielding an apparent heat of oxygenation delta H = -9.9 kcal/mol at pH = 8.0. The CO2 Bohr effect in group A was delta log P50/delta pH = -0.93 at 20 degrees C and -1.17 at 10 degrees C, whereas in group B no temperature effect was seen (-0.98 and -0.97). The acclimation temperature had no effect on the electrophoretic Hb pattern. As expected, the in vivo pH changed inversely with temperature from 8.06 at 10 degrees C to 7.73 at 20 degrees C, enhancing the temperature-induced shift in P50. Acclimation reverses partly the changes in O2 affinity, thereby improving the uptake of oxygen in the gills.

Acclimatization↗

Role of the carotid bodies in ventilatory acclimation to chronic hypoxia by the awake cat.

Steady-state breathing patterns during air and hypoxia (PIO2 = 84 Torr) were measured in awake cats in the following conditions: (1) during 7 months of exposure to air following carotid body resection (CBR; N = 6); (2) during 7 months of hypobaric hypoxia (PIO2 = 84 Torr; N = 5) following CBR; (3) during 5 months of exposure to hypobaric hypoxia (N = 4) while intact and then following CBR. Also, in groups (1) and (2) the aortic nerves were sectioned (ANX) at the end of the acclimation periods. The results show that the awake cat hypoventilates if the carotid bodies have been removed, and hypoxic sensitivity is reduced during long-term exposures to either hypoxia or normoxia. ANX caused a slight increase in respiratory frequency, indicating a minor role for the aortic bodies. CBR after acclimation to hypoxia resulted in decreased tidal volume but no change in respiratory frequency. The slight ventilatory acclimation to hypoxia in CBR cats was solely due to increased respiratory frequency. The phenomenon of 'hypoxic tachypnea' was modulated by acclimation, indicating that the effect of hypoxic acclimation upon respiratory frequency is due to central mechanisms.

Adaptation, Physiological↗

Cold acclimation and endurance training in guinea pigs: changes in lung, muscle and brown fat tissue.

The effects of an intermittent high intensity stimulus (running) or a chronic low intensity stimulus (cold acclimation) of oxidative metabolism on maximal oxygen uptake (VO2,max), lung O2 diffusing capacity (DLO2) and skeletal muscle as well as fat tissue mitochondrial content in growing guinea pigs are described. Young male guinea pigs were assigned to three experimental groups (n = 5): control (C), endurance trained (T; at 70% VO2max) or cold acclimated (CA; 5-7 degrees C) for six weeks. Animals were sacrificed at the end of the experimental period and tissue for morphometric analysis of the lung, muscle and interscapular fat was sampled. T and CA animals significantly increased weight specific VO2max by 23% and 29%, respectively. Despite a significant increase in absolute lung volume in T (+10%) and in weight specific lung volume in CA (+20%) neither absolute nor weight specific DLO2 was significantly affected by the experimental treatments. In trained animals the total volume of mitochondria remained unchanged in samples representative for the entire musculature but was significantly increased in M. vastus intermedius (+72%). Intramyocellular lipids increased significantly both in M. vastus intermedius (+244%) as well as in the whole body musculature (+164%). Cold acclimation increased the mitochondrial content of the interscapular fat pad by approximately 20-fold but had no effect on total mitochondrial volume in muscle. We conclude that the increase in oxygen demand resulting from exercise training or from cold acclimation could be accomodated by the existing lung diffusing capacity and did not induce a global change of oxidative capacity in skeletal muscle tissue in growing guinea pigs. Exercise training caused oxidative capacity to increase only in a locomotor muscle activated during running whereas cold acclimation greatly increased interscapular fat tissue oxidative capacity.

Acclimatization↗

Preexposure temperature acclimation and diet as modifying factors for the tolerance of golden ide (Leuciscus idus melanotus) to short-term exposure to 4-chloroaniline.

The influence of different temperature and nutrition regimes on the acute toxicity of 4-chloroaniline to golden ide (Leuciscus idus melanotus) was investigated. Acute toxicity was determined over 48 hr at 20 degrees C without feeding after a 2-day acclimation period. In an attempt to reveal underlying mechanisms accounting for diet- and temperature-related differences in the toxicant resistance of golden ide, biochemical and quantitative morphological parameters of the liver, a central organ in the xenobiotic metabolism of fish, were recorded throughout the 12-week acclimation period. In cold-acclimated fish, acute toxicity of 4-chloroaniline was 40% higher than in fish acclimated to 20 degrees C. If compared to 20 degrees C, preacclimation to 14 degrees C was characterized by a lower specific growth rate, an increase of hepatic glycogen, and a decrease of body and liver lipid deposits. The organelle composition of hepatocytes was not significantly altered by temperature acclimation. For the nutrition experiment, commercially available diets A and B of similar crude composition were used. Acute toxicity of 4-chloroaniline was 60% lower with diet B than with diet A. If compared to diet B, diet A induced a higher specific growth rate and increased hepatocellular volume of endoplasmic reticulum and Golgi fields, whereas glycogen and lipid of the liver as well as body lipid contents were reduced. The toxicity of 4-chloroaniline was correlated with the development of the endoplasmic reticulum, the major site of biotransformation enzymes. A consistent correlation with lipid contents could not be established. Results illustrate not only that assay conditions during the actual test may profoundly interact with results of toxicity studies, but also that maintenance conditions before the test can have significant consequences on results. In order to improve reproducibility of the results of acute toxicity tests, more consideration should be given to the standardization of pretest maintenance conditions of fish.

Adaptation, Physiological↗

Fluorescent histochemical demonstration of catecholamines in brown adipose tissue from obese (ob/ob) and lean mice acclimated at different temperatures.

Fluorescent histochemistry was used to visualize catecholamines in brown adipose tissue (BAT) of lean and genetically obese mice after they had been acclimated at different temperatures. At all temperatures, strong catecholamine-dependent fluorescence, attributable to the sympathetic innervation, was seen around the blood vessels of BAT from both lean and obese animals. Additionally, catecholamine-dependent fluorescent varicosities, in direct contact with the adipocytes were seen in abundance in lean mice acclimated at 23 degrees, 13 degrees or 4 degrees C and in obese mice acclimated at 13 degrees C. This latter compartment was greatly reduced in lean mice acclimated at 33 degrees C and in obese mice acclimated at 23 degrees and 33 degrees C. Three acute treatments (pretreatment with a monoamine oxidase inhibitor; 24 h food deprivation; and short-term cold exposure followed by short-term warm exposure) all increased the varicose fluorescence associated with adipocytes in obese mice housed at 23 degrees C, which suggests that the low resting level in these animals is attributable, at least in part, to subthreshold concentrations of catecholamines in existing varicosities rather than the absence of sympathetic varicosities per se. These results are in accordance with the results from noradrenaline turnover studies which suggest that the difference in sympathetic nervous system (SNS) activity in BAT from lean and obese (ob/ob) mice is best demonstrated at normal environmental temperatures. The reduced SNS activity in BAT of obese mice (which our studies show to be at the 'cellular' level) is likely to be a major factor in their reduced non-shivering thermogenesis and resultant high efficiency of energy storage as previously suggested by other workers.

Acclimatization↗

Altered responsiveness to parasympathetic activation of submaxillary salivary gland in the heat-acclimated rat.

Submaxillary salivary gland responsiveness during the heat acclimation procedure (34 +/- 1 degree C) was studied in the rat. Gland responsiveness was evaluated by measuring saliva flow rate of the anesthetized animals following either parasympathetic nerve stimulation or i.v. application of pilocarpine. A thirty percent decrease in glandular responsiveness for the two modes of stimulation was measured during the first 10 days of acclimation. Following 60 days of acclimation recovery was observed. It is concluded that decreased responsiveness of the heat-acclimated gland is a glandular event. Increased saliva flow occurring at the initial phase of acclimation is apparently due to changes in the thermoregulatory center.

Acclimatization↗