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The incorporation of unsaturated fatty acids of the n-9, n-6, and n-3 families into individual phospholipids by isolated hepatocytes of thermally-acclimated rainbow trout, Salmo gairdneri.

Rates of incorporation of 1-14C-oleic (18:1n9), -linoleic (18:2n6), and -linolenic (18:3n3) acids into individual phosphatides were determined in isolated hepatocytes from cold (5 degrees C)- and warm (20 degrees C)-acclimated rainbow trout, Salmo gairdneri. Fatty acid incorporation into phosphatidylcholine (PC) exceeded that into all other phospholipids, but at assay and acclimation temperatures of 5 degrees C, incorporation into phosphatidylethanolamine (PE) was generally intermediate between that of PC and the remaining phosphatides. Specific radioactivities (ratios of percentage isotope incorporation-to-mole percentage of phosphatide) were consistently less than one for both PC and PE, and greater than one for phosphatidic acid (PA), lysophosphatidylcholine (LPC), phosphatidylserine (PS), and cardiolipin (CL). For PS, specific radioactivities were greater in cold- than warm-acclimated trout, and greater at 5 degrees C than 20 degrees C. Rates of oleate incorporation were generally higher, and rates of incorporation of 18:2 and 18:3 lower in cold- than warm-acclimated trout. Most phospholipids demonstrated a clear preference for the incorporation of 18:2 when assayed at 20 degrees C; however, at 5 degrees C the incorporation of 18:2 was reduced and 18:3 was generally the preferred substrate. A reduction in assay temperature from 20 degrees C to 5 degrees C also shifted the incorporation of 18:2 away from PC into PS and PA. These data were interpreted to indicate 1) a cold-induced activation of PS metabolism, possibly resulting in elevated levels of PE; 2) lower rates of general acyl group turnover in animals acclimated to 5 degrees C than 20 degrees C; 3) a specificity to the acclimation response that favors the incorporation at cold temperatures of polyunsaturated fatty acids, but not the parent acids from which they are derived; and 4) the participation of a deacylation-reacylation cycle in the metabolism of phospholipids, particularly at cold temperatures.

Adaptation, Physiological↗

Eicosanoid synthesis by warm- and cold-acclimated American bullfrog (Rana catesbeiana) brain.

Previous studies in bullfrogs have demonstrated the presence of leukotriene (LT)C4 binding sites in the brain. However, synthesis of eicosanoids by brain tissue has not been examined. Because prostaglandin (PG) synthesis differs in warm- and cold-acclimated bullfrog lung tissue, this study compared the synthesis of prostaglandins and leukotrienes in brains from warm-(22 degrees C) and cold-acclimated (5 degrees C) animals. Initial experiments determined that leukotriene and prostaglandin production rates were greatest during the initial 30 min time period. Therefore, tissues were incubated in Munsick's solution and gassed with 95% O2, 5% CO2 for 30 min. Media were analyzed by radioimmunoassay for LTC4, LTB4, PGE2, PGF2 alpha, TXB2, and 6-keto PGF1 alpha. In warm-acclimated bullfrog brains, production was as follows: LTC4 > PGE2 > 6-keto PGF1 alpha, thromboxane (TX)B2, LTB4, and PGF2 alpha. Brain tissues from cold-acclimated animals incubated at 22 degrees C produced significantly greater quantities of PGE2 and 6-keto PGF1 alpha than did brains from warm-acclimated animals. Stimulation of TXB2 levels was observed when the animal was stunned with a blow to the head prior to decapitation. Indomethacin, a cyclooxygenase inhibitor, decreased prostaglandin but not leukotriene synthesis. Epinephrine (4 x 10(-8) M), the amphibian sympathetic postganglionic neurotransmitter, stimulated leukotriene synthesis by brains from warm-acclimated bullfrogs, and the effect was blocked with the 5-lipoxygenase inhibitor MK-886 (5 x 10(-5) M). These results clearly indicate that the bullfrog brain synthesized both leukotrienes and prostaglandins. Further studies are necessary to determine their function in the amphibian central nervous system.

Adaptation, Physiological↗

Bioavailability, biodegradation, and acclimation of Tetrahymena pyriformis to 1-octanol.

Previous work has indicated that the ubiquitous freshwater ciliate Tetrahymena pyriformis acclimates to the presence of hydrophobic chemicals acting by nonpolar narcosis. Four explanations have been identified to explain this apparent acclimation: (1) genetic adaptation occurs resulting in a resistant population, (2) T. pyriformis quickly biodegrades hydrophobic chemicals resulting in a perceived acclimation response, (3) hydrophobic chemicals are not bioavailable, and (4) T. pyriformis contain an endogenous biochemical adaptation system which can quickly cause cellular changes resulting in acclimation. Results of biodegradation experiments indicated that the total extractable 1-octanol did not change over the duration of the experiments. Bioavailability experiments were performed using the solid-phase microextraction technique. Although there is a decrease in freely available concentrations of 1-octanol over a 2.5 log unit range of Tetrahymena population density, the freely available concentration is constant for the population densities used for population growth experiments. Genetic change is highly unlikely since acclimation occurs in less than the time required for one population division. It is hypothesized that the acclimation response seen in Tetrahymena results from partitioning of the chemical into the membrane followed by active changes in the membrane structure to restore homeostasis.

1-Octanol↗

Effect of acclimation and fixation temperatures on the number of lamellae and periodicity of myelin in fibres of the optic nerve of goldfish.

Previous findings from our laboratory have shown that the optic nerves of goldfish acclimated to different temperatures differ considerably in their glycerophospholipid composition. This paper describes changes in the morphology of the nerve with different acclimation and fixation temperatures. Optic nerves of 5 and 25 degrees C acclimated fish were excised and fixed at the temperature of acclimation, or at the reverse temperature, and the morphology observed by electron microscopy. Under all temperature conditions considered there is a statistically significant linear relationship between the radius of the axon and the number of myelin lamellae. However, the temperature of acclimation and fixation both influence the regression coefficients for this relationship, the higher the acclimation temperature the lower the coefficient and the higher the fixation temperature the higher the coefficient. The periodicity of the myelin also alters with these temperatures, being greater in the 25 degrees C fish than in the 5 degrees C ones. Myelin sheath thickness is also significantly greater in the 25 degrees C fish. These results are discussed in relation to observed changes in glycerophospholipid composition and conduction velocities.

Animals↗

Effects of heat acclimation of distance runners in a moderately hot environment.

Five distance runners (H groups) performed a 60 min bicycle exercise at a load of 60--70% VO2max in a moderately hot environment (Ta: 33.5 degree C, 60% RH). Following a period of heat acclimation with bench-stepping at a load equal to about 25--30% VO2max, in a hot environment (Ta: 45--50 degree C, 30--40% RH) for 9 days, the work test was repeated. Two control subjects (R) performed the same work tests with no heat acclimation. Heat acclimation increased performance time. Rectal temperature, mean skin temperature, heart rate, and Na+ concentrations in sweat were lower in H and, with one exception, sweat rate was higher after heat acclimation. All H subjects demonstrated that the linear relationship between sweat rate and rectal temperature was shifted to a lower temperature (threshold shift). This shift correlated with a lowering of resting rectal temperature. The magnitude of the reduction in those two temperatures due to heat acclimation was identical. The observed improvement of work performance in moderate heat following heat acclimation to a higher temperature is attributed to a more efficient thermoregulatory mechanism.

Acclimatization↗

Basic fibroblast growth factor (bFGF) contributes to the enlargement of brown adipose tissue during cold acclimation.

The contribution of basic fibroblast growth factor to brown adipose tissue (BAT) enlargement during cold acclimation was investigated using rat brown adipocytes in primary culture. After cold exposure (at 5 degrees C) for 28 days, the level of bFGF messenger ribonucleic acid (mRNA) in BAT of cold-acclimated rats was markedly increased with the increase in the BAT weight. In addition, the blood plasma from cold-acclimated rats considerably enhanced the expression of basic fibroblast growth factor mRNA in rat brown adipocytes. Likewise, the blood plasma from cold-acclimated rats significantly stimulated the growth of rat brown adipocyte precursor cells compared with that from warm-acclimated rats, whereas there was no difference of effect between the two blood plasmas on the growth of bovine capillary endothelial cells. Basic fibroblast growth factor, but not platelet-derived growth factor stimulated the growth of brown adipocyte precursor cells. The conditioned medium from brown adipocyte primary culture markedly stimulated the growth of bovine capillary endothelial cells and the effect was inhibited considerably by antibasic fibroblast growth factor antibody. These results suggest that some factors concerned with the growth of brown adipocyte precursor cells are present in the blood plasma from cold-acclimated rats, and that basic fibroblast growth factor produced by brown adipocytes may significantly contribute to BAT enlargement by autocrine mechanisms during cold exposure.

Acclimatization↗

Interaction between heat acclimation and exogenous insulin in brown adipose tissue of rats.

Seventy-one male Wistar strain rats (7 weeks old) were kept at 5, 25, or 34 degrees C, respectively, for 2 weeks with or without insulin administration. Insulin (Novo Lente MC) was given subcutaneously in a dose of 3.62 nmol/125 microliters saline per 100 g body weight. An apparent effect of insulin treatment was noted only in heat-exposed rats, resulting in a remarkable gain in interscapular brown adipose tissue (BAT) mass of heat-acclimated, insulin-treated rats in terms of weight or weight per unit body weight. The BAT from heat-acclimated, insulin-treated rats had significantly higher levels of protein, DNA, RNA, and triglyceride than BAT from heat-acclimated, saline-treated rats. Therefore, it seems likely that the growth of BAT in heat-acclimated, insulin-treated rats was mostly due to the anabolic effects of insulin. The uncoupling protein mRNA was, however, present in BAT of heat-acclimated, insulin-treated rats at rather a depressed level, explaining a corresponding decrease in cold tolerance. On the other hand, the expression of insulin receptor mRNA was attenuated in BAT of rats from all the insulin-treated groups, possibly due to the down-regulation of insulin. Thus, there appeared to be some linkage among BAT, heat acclimation, and insulin.

Acclimatization↗

Temperature-dependent deacylation of molecular species of phosphatidylcholine by microsomal phospholipase A2 of thermally acclimated rainbow trout, Salmo gairdneri.

Using the ratios of kinetic parameters, V/Km, the deacylation of different molecular species of 1-palmitoyl,2-acyl phosphatidylcholine via microsomal phospholipase A2 (PLA2) was studied in liver tissue of thermally acclimated rainbow trout (Salmo gairdneri). In general, PLA2 from fish acclimated to cold temperatures showed an order of preference for the acyl moieties of 18:1 greater than 18:2 greater than 18:0. Trout acclimated to warm temperatures generally preferred 18:0 PC, but the actual order of preference depended on the temperature of the assays and the presence of endogenous lipids in the enzyme preparation. At 5 C, the particulate (microsomal) enzyme preferred 18:0 greater than 18:2 greater than 18:1, but a lipid-free preparation of the enzyme preferred 18:2 greater than 18:0 greater than 18:1. At 20 C, particulate enzyme preferred 18:1 greater than 18:0 greater than 18:2 but purified enzyme preferred 18:0 greater than 18:2 greater than 18:1. Thus, assay temperature and the presence of microsomal lipids had a greater effect on PLA2 from fish acclimated to warm temperatures than fish acclimated to cold temperatures. The substrate preference of PLA2 is discussed with reference to the previously observed changes in membrane fatty acid composition that occur with thermal acclimation in rainbow trout.

Acclimatization↗

Cold-water acclimation does not modify whole-body fluid regulation during subsequent cold-water immersion.

We investigated the impact of cold-water acclimation on whole-body fluid regulation using tracer-dilution methods to differentiate between the intracellular and extracellular fluid compartments. Seven euhydrated males [age 24.7 (8.7) years, mass 74.4 (6.4) kg, height 176.8 (7.8) cm, sum of eight skinfolds 107.4 (20.4) mm; mean (SD)] participated in a 14-day cold-water acclimation protocol, with 60-min resting cold-water stress tests [CWST; 18.1 (0.1) degrees C] on days 1, 8 and 15, and 90-min resting cold-water immersions [18.4 (0.4) degrees C] on intervening days. Subjects were immersed to the 4th intercostal space. Intracellular and extracellular fluid compartments, and plasma protein, electrolyte and hormone concentrations were investigated. During the first CWST, the intracellular fluid (5.5%) and plasma volumes were reduced (6.1%), while the interstitial fluid volume was simultaneously expanded (5.4%). This pattern was replicated on days 8 and 15, but did not differ significantly among test days. Acclimation did not produce significant changes in the pre-immersion distribution of total body water, or changes in plasma osmolality, total protein, electrolyte, atrial natriuretic peptide or aldosterone concentrations. Furthermore, a 14-day cold-water acclimation regimen did not elicit significant changes in body-fluid distribution, urine production, or the concentrations of plasma protein, electrolytes or the fluid-regulatory hormones. While acclimation trends were not evident, we have confirmed that fluid from extravascular cells is displaced into the interstitium during acute cold-water immersion, both before and after cold acclimation.

Acclimatization↗

Identification and characterization of three novel cold acclimation-responsive genes from the extremophile hair grass Deschampsia antarctica Desv.

Deschampsia antarctica Desv. is the only monocot that thrives in the harsh conditions of the Antarctic Peninsula and represents an invaluable resource for the identification of genes associated with freezing tolerance. In order to identify genes regulated by low temperature, we have initiated a detailed analysis of its gene expression. Preliminary 2-D gels of in vivo-labeled leaf proteins showed qualitative and quantitative differences between cold-acclimated and non-acclimated plants, suggesting differential gene expression. Similarly, cold-acclimation-related transcripts were screened by a differential display method. Of the 38 cDNAs initially identified, three cDNA clones were characterized for their protein encoding, expression pattern, response to several stresses, and for their tissue-specific expression. Northern blot analysis of DaGrx, DaRub1, and DaPyk1 encoding a glutaredoxin, a related-to-ubiquitin protein, and a pyruvate kinase-like protein, respectively, showed a distinct regulation pattern during the cold-acclimation process, and in some cases, their cold response seemed to be tissue specific. All three transcripts seem to be responsive to water stress as their levels were up-regulated with polyethyleneglycol treatment. DaRUB1 and DaPyk1 expression was up-regulated in leaf and crown, but down-regulated in roots from cold-acclimated plants. The significance of these results during the cold-acclimation process will be discussed.

Acclimatization↗

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 fatty acid composition of both membrane preparations generally became more unsaturated at lower acclimation temperatures, though the differences between 5 degrees C and 25 degrees 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 diphynylhexatriene in mitochondrial and microsomal membranes were highly hindered, though the hindrance offered by membranes of 25 degrees C acclimated green sunfish was far greater than that offered by the membranes of 5 degrees C acclimated fish, thus supporting the concept of homeoviscous adaptation. The absolute rotational rate was not consistently affected by acclimation treatment.

Acclimatization↗

Role of endocrine pancreas in temperature acclimation.

Role of endocrine pancreas in temperature acclimation in rats was investigated. Plasma glucagon level increased and insulin level decreased in cold-acclimated rats (CA). The reverse was observed in heat-acclimated rats (HA). In the pancreas there were no changes in glucagon and insulin in CA, but a decrease in glucagon and an increase in insulin were found in HA. Plasma insulin/glucagon molar ratio (I/G) declined in CA and rose in HA. Pancreatic I/G rose in HA. Acute cold exposure elevated plasma glucagon, but did not effect plasma insulin. Pancreatic glucagon, insulin and I/G were not influenced by acute cold exposure, while plasma I/G decreased. Plasma I/G was inversely correlated with both blood free fatty acids and glucose levels. These results suggest that endocrine pancreas is closely associated with metabolic acclimation to cold and heat through its regulation of the metabolic direction to catabolic phase in cold acclimation and to anabolic phase in heat acclimation.

Acclimatization↗

Acute effects of fat and carbohydrate on metabolic rate in normal, cold-acclimated and lean and obese (fa/fa) Zucker rats.

The rise in metabolic rate after intragastric feeding with fat and carbohydrate was enhanced in cold-acclimated (5 degrees C) rats and diminished in warm-acclimated (30 degrees C) rats compared to controls (24 degrees C), but the response was largest in cold-acclimated animals intubated with fat. These acute effects of nutrients were almost completely abolished by beta-adrenergic blockade with propranolol in all groups, while the parasympathetic antagonist atropine sulphate enhanced the responses in control rats, but had little effect in cold-acclimated animals. Feeding carbohydrate produced similar increases in interscapular brown adipose tissue (BAT) temperature in control and cold-acclimated rats, but fat caused a much greater rise in the latter group. The thermic effects of both nutrients were lower in genetically obese Zucker rats than in their lean littermates. Atropine slightly increased the thermic responses to fat and carbohydrate in the lean Zucker rats and caused marked potentiation in obese rats intubated with fat, but did not alter the effect of carbohydrate in the obese animals These data suggest that the size of the acute rise in metabolic rate after fat and carbohydrate is dependent on the thermogenic capacity of the animal. The response to fat was particularly large in cold-acclimated rats, where BAT activity is high, possibly due to a direct action of fat on the tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

Acclimatization↗

Sympathetic outflow to interscapular brown adipose tissue in cold acclimated mice.

C57BL/6J male mice were subjected to a cold acclimation procedure which consisted of three consecutive cold stress tests: 3-h partial restraint at 6 degrees C at 2-wk intervals. During the week following the last cold stress test, each animal previously subjected to the cold acclimation procedure, and an additional group of naive mice (animals that never had been exposed to an environment below room temperature) were anesthetized with urethan, paralyzed with vecuronium bromide, artificially ventilated, and subjected to cold stimulation for approximately 16 min. Electrical impulse activity from one of the fine sympathetic nerves entering the interscapular brown adipose tissue was recorded before and during cold stimulation, until body temperature dropped 8 degrees C below control level. Sympathetic outflow increased significantly during cold stimulation in all mice. Animals that did not achieve cold acclimation in three repeated cold stress tests (they demonstrated less cold tolerance in the last test) had lower sympathetic nervous outflow to brown adipose tissue at room temperature and during cold stimulation than mice that had achieved cold acclimation. In fact, sympathetic nervous outflow to brown adipose tissue in mice that had failed to show cold acclimation was similar to that of naive mice. These findings indicate that the sympathetic nervous system plays a primary role in cold acclimation.

Acclimatization↗

Temperature acclimation of respiratory function in the salamander Taricha granulosa.

The effect of acute and chronic temperature change on oxygen uptake (VO2) and the respiratory properties of blood was determined in the newt, Taricha granulosa. Acclimation to 20-24 degrees C for 4 weeks caused a significant increase in hemoglobin concentration (6.5-9.5 g/dl) and O2 capacity of blood but no change in oxygen affinity compared with animals acclimated to 4-6 degrees C. Cold acclimated animals had a reverse Bohr effect compared with warm acclimated animals (dlogP 50/dpH = +0.13 and -0.12, respectively) but the present data offer no mechanism. There was no difference between the two acclimation groups with respect to temperature sensitivity of O2 binding, cooperativity of binding, buffer capacity, red cell organic phosphate concentration, or red cell dimensions. There was no evidence for thermal acclimation of VO2. Over the seasonal range of water temperature experienced by this species (5-25 degrees C), the major adaptation to the ca. 4 fold increase in VO2 is a 49% increase in blood O2 capacity.

Acclimatization↗

Cold acclimation and endurance training in guinea pigs: changes in daily and maximal metabolism.

The physiological effects of training or cold acclimation on maximal oxygen uptake (VO2,max) and average daily metabolic rate (VO2,dav) of a small mammal, the guinea pig, are described. Young male guinea pigs were assigned to three experimental groups; control, endurance trained (70% VO2,max) or cold acclimated (5-7 degrees C) for six weeks. Measurements of VO2,max and VO2,dav were made before and after the treatments. VO2,max increased significantly in cold acclimated (+29%) and endurance trained (+23%) animals and was achieved at a higher maximal running speed compared to post-treatment controls. Maximal blood lactate concentration was significantly higher in cold acclimated compared to endurance trained animals. Endurance trained animals had a reduced VO2,dav compared to control animals, whereas cold acclimation raised VO2,dav in the cold as expected, but also at room temperature. All three groups showed a daily pattern in metabolic rate (night > day). In conclusion, both endurance training and cold acclimation lead to enhanced VO2,max and changes in resting oxygen consumption throughout the day.

Acclimatization↗

Temporal induction pattern of hepatic cytochrome P450 1A in thermally acclimated dab (Limanda limanda) treated with 3,3',4,4'-tetrachlorobiphenyl (CB77).

Mature male dab (Limanda limanda) acclimated at 10 degrees and 16 degrees C were orally administered a single dose of 0.5 mg/kg 3,3',4,4'-tetrachlorobiphenyl (CB77). At both temperatures, levels of cytochrome P450 1A (CYP1A) protein and 7-ethoxyresorufin O-deethylase (EROD) activity showed a two to six fold induction 40 days after CB77 treatment compared to control groups. Maximum responses of both EROD activity and CYP1A protein for the warm-acclimated fish were observed at 5 days after treatment. For the cold-acclimated fish a slow, progressive elevation for both EROD activity and CYP1A protein was observed and maximum responses were measured 40 days after treatment. Absolute EROD activity and CYP1A protein levels of fish from both temperatures were equally high at 40 days after treatment. Since in the control groups EROD activity and CYP1A protein levels were higher in the cold-acclimated fish, the magnitude of induction was higher in the warm acclimated ones. The highest concentrations of CB77 in muscle of fish from both temperatures were found at 5 and 10 days after treatment. The liver somatic index (LSI) showed 1.5 fold significantly higher values for the fish acclimated at 10 degrees C.

Administration, Oral↗

Use of helium-oxygen to examine the effect of cold acclimation on the summit metabolism of a marsupial, Dasyuroides byrnei.

To determine whether marsupial mammals increase their metabolic capabilities during cold acclimation, the metabolism of both warm and cold acclimated Dasyuroides byrnei was examined by exposure to cold in a helium-oxygen atmosphere. Mean values of heat production and conductance were significantly higher in a helium-oxygen atmosphere than in air. Body temperature did not change until metabolic capacity was exhausted. Both cold and warm acclimated groups could maintain a metabolic scope of 10-11 times the basal or standard level for this species. Such a metabolic scope is much higher than levels recorded for placental mammals. At very low ambient temperatures cold acclimated D. byrnei could sustain a high level of heat production longer than could warm acclimated animals. While there are some similarities between marsupial mammals and placental mammals in their responses to cold acclimation, an increase in maximum metabolism, as reported for placentals, does not seem to occur in marsupials.

Acclimatization↗