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Lipids as energy source during salinity acclimation in the euryhaline crab Chasmagnathus granulata dana, 1851 (crustacea-grapsidae).

Lipids seem to be the major energy store in crustaceans. Moreover, they are extremely important in maintaining structural and physiological integrity of cellular and sub cellular membranes. During salinity adaptation, energy-demanding mechanisms for hemolymph osmotic and ionic regulation are activated. Thus, the main goal of this work was to verify the possible involvement of lipids as an energy source in the osmotic adaptation process. The estuarine crab Chasmagnathus granulata was captured and acclimated to salt water at 20 per thousand salinity and 20 +/- 2 degrees C, for 30 days. After acclimation, crabs were divided into groups of ten and transferred to fresh water (0 per thousand ), salt water at 40 per thousand salinity, or maintained in salt water at 20 per thousand salinity (control group), without feeding. Before and seven days after the salinity change, wet weight and lipid concentration in gills, muscle, hepatopancreas, and hemolymph were determined according to the colorimetric assay of sulphophosphovanilin. Results show that hepatopancreas lipids were not mobilized during osmotic stress regulation. Gill and muscle lipids were significantly lower in crabs subjected to hypo-osmotic stress than those subjected to the hyper-osmotic stress or maintained at the control salinity. Our results point to the occurrence of lipid mobilization and involvement of these compounds in the osmotic acclimation process in C. granulata, but with differences between tissues and the osmotic shock (hypo or hyper) considered.

Adaptation, Physiological↗

Effects of cold acclimation on the activity levels of creatine kinase, lactate dehydrogenase and lactate dehydrogenase isoenzymes in various tissues of the rat.

The effects of cold acclimation on the activity levels of creatine kinase, lactate dehydrogenase and lactate dehydrogenase isoenzymes in various tissues/ organs of the rat (Rattus norvegicus) were investigated. Male Sprague-Dawley rats were divided into two groups. One group was housed at 4+/-1 degrees C (experimental group) and the other at 24+/-1 degrees C (control group) for six months. The rats were housed in single cages and had access to food and water ad libitum. The tissues/organs investigated were heart, liver, lung, kidney, gastrocnemius muscle and interscapular brown adipose tissue as well as serum. With the exception of lung, (which showed a decrease of 24%) total creatine kinase activity levels were significantly increased (P< 0.05) in all the tissues/organs investigated (17-51%) as well as serum (34%), in cold acclimated animals. Cold acclimation also resulted in significantly increased (P< 0.05) activity levels of lactate dehydrogenase in all the tissues/organs investigated (14-24%) as well as serum (35%). Cold exposure resulted in an increase of the activity levels of all the detectable isoenzymes of lactate dehydrogenase, although not always significant, in all the tissues/organs investigated as well as serum. The M(4)tetramer of lactate dehydrogenase was the only detectable isoenzyme in serum.

Acclimatization↗

Active oxygen scavengers during cold acclimation of Scots pine seedlings in relation to freezing tolerance.

Freezing injury of plants may be caused by the deleterious reactions of active oxygen species, and free-radical scavenging systems may be important in the alleviation of freezing stress. To test the feasibility of this hypothesis, enzymes and metabolites that cooperatively scavenge O2 and H2O2 were analyzed in Scots pine (Pinus sylvestris L.) seedlings during a stepwise cold acclimation procedure. Elevated levels of enzymatic scavengers such as ascorbate peroxidase, glutathione reductase, monodehydroascorbate reductase, and dehydroascorbate reductase were found, along with increased freezing tolerance during cold acclimation, supporting the hypothesis. Induction of the scavenging systems during acclimation is discussed in relation to freezing tolerance.

Acclimatization↗

Antibody-producing cells correlated to body weight in juvenile chinook salmon (Oncorhynchus tshawytscha) acclimated to optimal and elevated temperatures.

The immune response of juvenile chinook salmon (Oncorhynchus tshawytscha) ranging in weight from approximately 10 to 55 g was compared when the fish were acclimated to either 13 or 21 degrees C. A haemolytic plaque assay was conducted to determine differences in the number of antibody-producing cells (APC) among fish of a similar age but different body weights. Regression analyses revealed significant increases in the number of APC with increasing body weight when fish were acclimated to either water temperature. These results emphasise the importance of standardising fish weight in immunological studies of salmonids before exploring the possible effects of acclimation temperatures.

Acclimatization↗

Characterization of adrenoceptor types and subtypes in American bullfrogs acclimated to warm or cold temperature.

While indirect evidence suggested that the responsiveness of frog adrenoceptors changes in response to temperature, direct measurement of adrenoceptor binding following acclimation to warm and cold temperatures had not been done. In the present study, the radioligands [3H]prazosin, [3H]RX821002, and [125I]cyanopindolol were used to label and quantify alpha 1-, alpha 2-, and beta-adrenoceptors in bullfrogs acclimated to warm or cold environments. The number of alpha 1-, alpha 2-, and beta-adrenoceptors in atrium, ventricle, and kidney membranes was not significantly different between warm- and cold-acclimated frogs. Characterization of receptor subtypes using pharmacological antagonists demonstrated that alpha 2-adrenoceptors in frog spinal cord and kidney were of the same pharmacological subtype, which is similar to the mammalian alpha 2A-subtype. The beta-adrenoceptor in frog ventricle, atrium, and kidney was the beta 2-subtype. These results suggest that while the alpha 1-, alpha 2-, and beta-adrenoceptor types have evolved in the frog, multiple subtypes of adrenoceptors are not necessary for physiological regulation in this species.

Acclimatization↗

Localization and changes in distribution of brain alpha 2- and beta-adrenoceptors in response to acclimation state in the American bullfrog (Rana catesbeiana).

Alpha (alpha)- and beta (beta)-adrenoceptors regulate physiological processes in vertebrates. This study determined the location of alpha 2- and beta-adrenoceptors in the brain of the American bullfrog, Rana catesbeiana, using autoradiography. As the density of receptors may be affected by environmental temperature, a comparative numerical analysis of adrenoceptors in the areas of localization with respect to warm and cold acclimation was also carried out. Areas of greatest concentration of alpha 2-adrenoceptors were the accessory olfactory bulb, medial pallium, and olfactory bulb. Adrenoceptor numbers were significantly decreased in the accessory olfactory bulb and medial pallium in cold-acclimated animals. beta-adrenoceptors were localized in the thalamus, cerebellum, medial pallium, and amygdala/ striatum. Cold acclimation decreased adrenoceptor density in medial pallium and torus semicircularis and increased adrenoceptor density in the thalamus and hypothalamic preoptic areas. Among the alpha 2- and beta-adrenoceptors, only four regions of overlap existed, the medial pallium, hypothalamic preoptic area, optic tract, and isthmic tegmentum. Otherwise, where there were alpha 2-adrenoceptors, there were few or no beta-adrenoceptors. No alpha 2- or beta-adrenoceptors were found in the pituitary and optic chiasm. The distribution of adrenoceptors in particular areas of the brain may have functional significance with respect to physiological changes which occur in response to hibernation.

Acclimatization↗

Cold acclimation and cold-regulated gene expression in ABA mutants of Arabidopsis thaliana.

We have examined the cold-induced enhancement of freezing tolerance and expression of cold-regulated (cor) genes in Arabidopsis thaliana (L.) Heynh (Landsberg 'erecta') and abscisic acid (ABA)-deficient (aba) and ABA-insensitive (abi) mutants derived from it. The results indicate that the abi mutations had no apparent effect on freezing tolerance, while the aba mutations did: cold-acclimated aba mutants were markedly impaired in freezing tolerance compared to wild-type plants. In addition, it was observed that non-frozen leaves from both control and cold-treated aba mutant plants were more ion-leaky than those from corresponding wild-type plants. These data are consistent with previous observations indicating that ABA levels can affect freezing tolerance. Whether ABA has a direct role in the enhancement of freezing tolerance that occurs during cold acclimation, however, is uncertain. Several studies have suggested that ABA might mediate certain changes in gene expression that occur during cold acclimation. Our data indicate that the ABA-induced expression of three ABA-regulated Arabidopsis cor genes was unaffected in the abi2, abi3, and aba-1 mutants, but was dramatically impaired in the abi1 mutant. Cold-regulated expression of all three cor genes, however, was nearly the same in wild-type and abi1 mutant plants. These data suggest that the cold-regulated and ABA-regulated expression of the three cor genes may be mediated through independent control mechanisms.

Abscisic Acid↗

Effects of heat acclimation on the secretory products of the submaxillary gland of the rat. An ultrastructural and biochemical study.

Secretory granule area and glycoprotein concentration of the saliva in the submaxillary gland of rats were measured during various stages of acclimation to heat at 34 +/- 1 degrees C. Granule size decreased by 18% during the first five days of heat acclimation (0.025 less than p less than 0.05) after which period it increased to reach 118% of the control levels after 28 days (p less than 0.05). Glycoprotein concentration in the saliva of stimulated glands rose above control levels, reaching a maximum between the 2nd and 5th day of acclimation (p less than 0.05). It was concluded that the initial decrease in granule size reflects a decrease in glycoprotein content following an increase in salivary flow known to occur at high ambient temperatures. The subsequent increase in granule size is considered an adaptation of the gland to continuous stimulation. The rise in salivary glycoprotein concentration suggests increased efficiency of the secretory mechanism.

Adaptation, Physiological↗

Inactivation of brown adipose tissue thermogenesis by oestradiol treatment in cold-acclimated rats.

Both cold-acclimated female rats and rats at thermoneutrality received 0.15-0.20 mg daily of 17 beta-oestradiol over 15 days via a Silastic capsule implanted subcutaneously. Controls received empty implants. Comparison between the oestradiol-treated animals and the untreated controls revealed that at thermoneutrality, oestradiol treatment decreased food intake and body weight gain, but did not affect brown adipose tissue (BAT) thermogenesis and composition. By contrast, in cold-acclimated rats, oestradiol treatment did not modify food intake or body weight gain, but it decreased BAT thermogenesis. It is concluded that the effects of oestradiol treatment on BAT depend on the activity of the tissue, i.e. it has no effect on BAT when the tissue is thermogenically inactive, but it decreases cold-induced BAT thermogenesis. It is suggested that oestradiol could be the hormonal factor responsible for the previously observed inactivation of BAT thermogenesis during pregnancy in cold-acclimated rats.

Adipose Tissue↗

Effect of hidromeiosis on sweat drippage during acclimation to humid heat.

Sweat rate and the rate of change in sweat drippage were studied during the acclimation of eight healthy male subject during exposure to heat during 10 consecutive days. During acclimation to hot humid conditions, the increase in total body sweat rate results in an increase in the rate of sweat drippage. We found, however, that on each day the drippage rate markedly decreased with time after the 1st h of heat exposure. This hidromeiosis was investigated as a function of the heat exposure time. No shortening of the onset time of hidromeiosis occurred with acclimation. With repeated heat exposures, the initial sweat rates in response to stress increased, and the subsequent decline became larger with higher sweat rates at the time of onset of hidromeiosis. Hidromeiosis appears to be a function of the degree of skin wettedness reached in the various local skin areas which determine the overall body skin wettedness upon which evaporative adjustments depend. Thus, the observed overshoot in total sweat rate as indicated by sweat drippage, and the subsequent hidromeiosis, result from initial oversweating in the poorly ventilated areas of skin. This sweat decline seems to be due to a reduction in output of the active sweat glands rather than to a reduction in active sweat gland number.

Acclimatization↗

Modifications of sweating responses to thermal transients following heat acclimation.

The sweating response was studied before and after passive humid heat acclimation in four resting male subjects who were exposed to slow thermal transients increasing air and wall temperatures from 28 degrees C to 45 degrees C. The slopes of the ambient temperature increases were +0.19 degrees C . min-1; +0.16 degrees C . min-1 or +0.14 degrees C . min-1. Dew-point temperature and air velocity were kept constant (17.5 degrees C; 0.3 m . s-1). Continuous measurements were made of oesophageal temperature, mean skin temperature, whole-body sweat loss and of right upper limb sweating responses. The local sweating response was measured from an arm chamber under a local thermal clamp (Tsk,1 = 38 degrees C). The results confirmed the fact that heat acclimation to humid heat induces a shortening in the time lag of sweat onset and increases the local sweating rates while internal temperature changes are reduced. These modifications are interpreted as a non-linearity in the response of the central controller, involving both a change in the central gain and an upward resetting of the "local sweating rate-body temperature" curves, without any shifting of the hypothalamic set-point temperature as it is currently described. However, a modification of local sweat gland activity occurring with heat acclimation cannot be ruled out.

Acclimatization↗

Interaction between exercise training and cold acclimation in rats.

Five groups of 10 rats were used. Group A included sedentary rats kept at 24 degrees C, group B exercised-trained rats and group C rats exposed at -15 degrees C for 2 h every day and kept at 24 degrees C for the remaining time. These 3 groups were kept on this regimen for 10 weeks. In addition group D was acclimated to cold (2 h.d-1 at -15 degrees C) for 6 weeks and subsequently deacclimated at 24 degrees C for 4 weeks. Group E was also acclimated to cold for 6 weeks and during the deacclimation, at 24 degrees C period which lasted 4 weeks, the animals were exercised 2 h per day. Following the 10 week experimental period all animals were sacrificed and DNA and protein content of the IBAT as well as its total mass were measured. The results show significant increases in the cold adapted group. Exercise training which had no effect on brown adipose tissue IBAT at room temperature, caused an accelerated reduction in weight, DNA and protein content of the BAT in rats previously acclimated to cold. In spite of this, the thermogenic response to noradrenaline was significantly enhanced in the group which exercised during the deacclimation period. It is suggested that tissues other than IBAT may explain this enhanced heat production capacity.

Acclimatization↗

Physiological significance of catalase and glutathione peroxidases, and in vivo peroxidation, in selected tissues of the toad Discoglossus pictus (Amphibia) during acclimation to normobaric hyperoxia.

1. Various parameters related to oxidative stress were measured in adult Discoglossus pictus acclimated for 15 days to either normoxia or hyperoxia (PO2 = 710 mmHg). 2. Total weight of the toads and total and relative wet weight of liver, kidneys, lungs and heart were not changed by hyperoxic acclimation. 3. In vivo tissue peroxidation increased in lung, decreased in skeletal muscle, and was not changed in liver, kidney, heart and skin after hyperoxic exposure. 4. Hyperoxic acclimation increased catalase activities in the lung, liver, kidney and heart but not in skeletal muscle and skin. 5. Liver showed higher GSH-peroxidase activity with cumene-OOH than with H2O2 as substrate, whereas lung, skeletal muscle and skin presented similar GSH-peroxidase activities with both substrates. 6. GSH-peroxidase activities did not change between hyperoxic and normoxic animals in liver, lung, skeletal muscle and skin. 7. These results show that catalase, not GSH-peroxidase, is the principal H2O2 detoxifying enzyme involved in the adaptation of D. pictus to hyperoxia.

Adaptation, Physiological↗

Blood flow and muscle bio-energetics by 31P-nuclear magnetic resonance after local cold acclimation.

To clarify the origin of local cold adaptation and to define precisely its influence on muscle bio-energetics during local exercise, five subjects were subjected to repeated 5 degrees C cold water immersion of the right hand and forearm. The first aim of our investigation was therefore carried out by measuring local skin temperatures and peripheral blood flow during a cold hand test (5 degrees C, 5 min) followed by a 10-min recovery period. The 31P by nuclear magnetic resonance (31PNMR) muscle bio-energetic changes, indicating possible heat production changes, were measured during the recovery period. The second aim of our investigation was carried out by measuring 31PNMR muscle bioenergetics during handgrip exercise (10% of the maximal voluntary contraction for 5 min followed by a 10-min recovery period) performed both at a comfortable ambient temperature (22 degrees C; E) and after a cold hand test (EC), before and after local cold adaptation. Local cold adaptation, confirmed by warmer skin temperatures of the extremities (+30%, P less than 0.05), was related more to an increased peripheral blood flow, as shown by the smaller decrease in systolic peak [-245 (SEM 30) Hz vs -382 (SEM 95) Hz, P less than 0.05] than to a change in local heat production, because muscle bioenergetics did not vary. Acute local cold immersion decreased the inorganic phosphate:phosphocreatine (PC) ratio during EC compared to E [+0.006 (SEM 0.010) vs +0.078 (SEM 0.002) before acclimation and +0.029 (SEM 0.002) vs +0.090 (SEM 0.002) after acclimation respectively, P less than 0.05] without significant change in the PC:beta-adenosine triphosphate ratio and pH. Local adaptation did not modify these results statistically. The recovery of PC during E increased after acclimation [9.0 (SEM 0.2) min vs 3.0 (SEM 0.4) min, P less than 0.05]. These results suggested that local cold adaptation is related more to peripheral blood flow changes than to increased metabolic heat production in the muscle.

Acclimatization↗

Seasonal acclimation of bank voles and wood mice: nonshivering thermogenesis and thermogenic properties of brown adipose tissue mitochondria.

Seasonal acclimation of nonshivering thermogenesis and brown adipose tissue was studied in wild bank voles (Clethrionomys glareolus), yellow necked field mice and wood mice (Apodemus flavicollis, A. sylvaticus). Both, voles and mice increased their capacity for nonshivering thermogenesis during winter. Thermogenic properties of brown fat (cytochrome c oxidase activity, mitochondrial protein content, GDP-binding of brown fat mitochondria) showed similar changes during seasonal acclimation; Clethrionomys and Apodemus spp. both showed lowest thermogenic properties in the summer during August, a rapid increase during fall, and highest levels of thermogenic activity in the winter months. With regard to changes in body weight and brown fat mass these species show different strategies for seasonal acclimation. In Clethrionomys a reduction of body mass in the winter was found, both in the wild population as well as in individual animals housed in the laboratory. A. flavicollis showed a reduction of body weight during fall, whereas A. sylvaticus maintained a constant body mass throughout the year. Brown fat mass and cellularity increased in the Apodemus spp. during winter, in parallel with the thermogenic properties of brown fat, whereas in Clethrionomys brown fat mass and cellularity remained seasonally constant. These species live in the same habitat and were trapped in the same area. It is concluded that seasonal improvements of in vivo and in vitro thermogenesis are very similar in these species, although the physiological basis for this improvement is different in Clethrionomys and Apodemus.

Acclimatization↗

Parallel measurements of heat production and thermogenin content in brown fat cells during cold acclimation of rats.

The maximum thermogenic capacity of brown fat cells from control and cold acclimated rats was measured using a continuous-flow microcalorimetric system. The content of the 32.000 D, brown fat specific protein, thermogenin, was measured in the cells used for heat production measurements by competitive ELISA. The ratio between the maximal thermogenic capacity and the amount of thermogenin for control and cold acclimated rats was compared. It was found that the ratio between the two parameters decreased during cold acclimation due to a decrease in maximal thermogenic capacity and an increase in the amount of thermogenin, indicating regulation of heat production either at thermogenin or receptor level.

Acclimatization↗

Effects of cold exposure on cyclic AMP concentration in plasma, liver, and brown and white adipose tissues in cold-acclimated rats.

Effects of acute cold exposure on plasma energy substrates and tissue 3',5'-adenosine monophosphate (cAMP) were analyzed in intact rats, to define an involvement of the nucleotide in nonshivering thermogenesis (NST) and resultant cold acclimation. After an acute cold exposure to -5 degrees C, the plasma glucose level increased gradually in warm-kept control rats (C) while it decreased significantly in cold-acclimated rats (CA). However, it was increased considerably by an extreme cold exposure to -15 degrees C in both C and CA. By contrast, plasma levels of free fatty acids (FFA) increased immediately after cold exposure and the release lasted during the period of exposure especially in C. The cold exposure also increased plasma cAMP concentration but no concomitant increase was found in the liver. In both brown (IBAT) and white (WAT) adipose tissues the nucleotide concentration showed a stepwise decrease. The observed correlation between lipolysis and plasma cAMP response after cold exposure suggests an involvement of the adenylate cyclase-cAMP system in NST via lipid metabolism, at least, in the early stages of cold acclimation.

Acclimatization↗

Quantitative evaluation of gap junctions in rat brown adipose tissue after cold acclimation.

The decrease in the metabolic capacity of rat brown adipose tissue during the late postnatal period can be reversed by cold acclimation of the animals. In order to find out whether a parallel decrease in capability for intercellular communication observed during this period is also reversed by cold acclimation, gap junction size and number per unit area of cell surface have been quantified in freeze-fracture replicas; cell diameters have been measured in semi-thin sections. It was found that the specific number of gap junctions remains unchanged during cold acclimation. However, the mean gap junction size increases by 75% and the ratio of gap junctional area per cell volume, an index for intercellular exchange capacity, is doubled. This result illustrates further the parallelism between metabolic capacity and cell communication in brown fat.

Acclimatization↗