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Induction of Cold Acclimation in Cornus stolonifera Michx.

A warm (20 to 15 Celsius day or night) preconditioning treatment enhanced cold acclimation of Cornus stolonifera bark under short-day conditions when plants were preconditioned for at least 4 weeks. Warm preconditioning inhibited the acclimation of plants subjected to long photoperiods. Removing leaves from plants exposed to low temperatures and short days inhibited acclimation. Removal of buds did not affect acclimation. Plants did not acclimate unless they were exposed to at least 4 weeks of short photoperiods prior to defoliation. Plants began to acclimate to cold at the time of growth cessation but not before. When half of the leaves were removed from plants, the defoliated and foliated branches both acclimated as well as branches on completely foliated plants. Girdling the phloem between foliated and defoliated branches prevented acclimation of the latter regardless of the position of the girdle in relation to the root system and the defoliated branch. When all of the leaves of plants were covered with aluminum foil to exclude light after 0 or 4 weeks of exposure to short days, the results resembled a defoliation study, i.e., plants with leaves covered at the start of the experiment failed to acclimate, and those covered after 4 weeks acclimated to some extent but less than uncovered control plants. Under longday conditions plants with all leaves covered failed to acclimate, and plants with none or half of their leaves covered acclimated equally and to a limited extent. Under short-day conditions, however, the covered branches of partially covered plants acclimated more than their uncovered counterparts or branches of totally uncovered plants.

Journal Article↗

Physiological responses of horses to a treadmill simulated speed and endurance test in high heat and humidity before and after humid heat acclimation.

To investigate whether horses were able to acclimate to conditions of high temperature and humidity, 5 horses of different breeds were trained for 80 min on 15 consecutive days on a treadmill at 30 degrees C and 80%RH. Training consisted of a combination of long duration low-intensity exercise, medium duration medium intensity exercise and short duration high intensity exercise. Between training sessions the horses were maintained at 11+/-3 degrees C and 74+/-2%RH. Before (PRE-ACC) and after acclimation (POST-ACC) the horses undertook a simulated Competition Exercise Test (CET), designed to represent the Speed and Endurance Test of a 3-day event, at 30 degrees C/80%RH. Maximal oxygen uptake (VO2PEAK) was not changed following acclimation (PRE-ACC 141+/-8 ml/min/kg bwt vs. POST-ACC 145+/-9 ml/min/kg bwt [STPD], P>0.05). Following acclimation, 4 of the 5 horses were able to complete a significantly greater amount of Phase D in the CET (PRE-ACC 6.3+/-0.3 min vs. POST-ACC 7.3+/-0.3 min, P<0.05; target time = 8 min). Resting body temperatures (pulmonary artery [TPA], rectal [TREC] and tail-skin [TTSK] temperatures) were all significantly lower following acclimation. During exercise, metabolic heat production (M) and heat dissipation (HD), for the same exercise duration, were both significantly lower following acclimation (P<0.05), although heat storage (HS) was significantly higher (P<0.05). The higher heat storage following acclimation was associated with a lower TTSK for a given TPA and a decreased total fluid loss (% bodyweight, P<0.05). Plasma volume was not changed following acclimation. The relationship of sweating rate (SR) to TPA or TTSK on either the neck or the gluteal region was not significantly altered by acclimation, although the onset of sweating occurred at a lower TPA or TTSK following acclimation (P<0.05). The horses in the present study showed a number of physiological adaptations to a period of 15 days of exposure to high heat and humidity consistent with a humid heat acclimation response. These changes were mostly similar to those reported to occur in man and other species and were consistent with thermal acclimation and an increased thermotolerance, leading to an improved exercise tolerance. It is concluded that a 15 day period of acclimation is beneficial for horses from cooler and or drier climates, that have to compete in hot humid conditions and that this may redress, to some extent, the decrement in exercise tolerance seen in nonacclimated horses and reduce the risk of heat related disorders, such as heat exhaustion.

Acid-Base Equilibrium↗

Phenotypic flexibility in the basal metabolic rate of laughing doves: responses to short-term thermal acclimation.

Many birds exhibit considerable phenotypic flexibility in maintenance energy requirements, and up- or downregulate basal metabolic rate (BMR) over time scales of days to weeks during thermal acclimation. However, the extent to which individual birds can reverse the direction of BMR adjustments over short time scales remains unknown. In this study, we examined metabolic responses to short-term thermal acclimation in laughing doves Streptopelia senegalensis. In 30 wild-caught doves (mean body mass=92.6 g) divided into three experimental groups of 10 birds each, initial BMR averaged 0.760+/-0.036 W. Thereafter, each group was acclimated to one of three acclimation air temperatures (T(acc)=10, 22 or 35 degrees C) for 21 days, during which time the doves were housed in individual cages. Following the first acclimation period (acclimation I), BMR (W) was significantly lower and was negatively and linearly related to T(acc) [BMR=0.714-0.005T(acc)]. Acclimation I BMR varied from 0.546+/-0.039 W in doves acclimated to T(acc)=35 degrees C to 0.665+/-0.058 W at T(acc)=10 degrees C. A second acclimation period of a further 21 days (acclimation II) revealed that the direction of BMR adjustments could be reversed within individuals, with acclimation II BMR again negatively and linearly related to T(acc). The slope of the relationship between BMR and T(acc) following acclimation II was not significantly different to that following acclimation I. BMR exhibited consistent inter-individual variation, with a low but significant repeatability of 0.113. The within-individual BMR variation of up to 26% that we observed in laughing doves reveals that BMR is a highly flexible trait in this species, and reiterates the need to take phenotypic plasticity into account in comparative analyses of avian energetic parameters.

Acclimatization↗

Changes in mitochondrial oxidative capacities during thermal acclimation of rainbow trout Oncorhynchus mykiss: roles of membrane proteins, phospholipids and their fatty acid compositions.

Changes in the properties of mitochondria from oxidative muscle of rainbow trout Oncorhynchus mykiss were examined during warm (5 degrees C to 15 degrees C) acclimation. Trout were studied shortly after the initial thermal change and after 8 weeks acclimation to 15 degrees C. To identify potential mechanisms by which oxidative capacities change, the modifications of phospholipid composition, membrane proteins and functional capacities of red muscle mitochondria were examined. Marked functional changes of isolated muscle mitochondria during warm acclimation of rainbow trout were reflected by a host of modifications in phospholipid composition, but by few shifts in protein components. Shortly after transfer of trout from 5 degrees C to 15 degrees C, the maximal oxidative capacity of mitochondria measured at 15 degrees C increased slightly, but rates at both assay temperatures (5 degrees C and 15 degrees C) decreased markedly after warm acclimation. The increase in capacity in short-term warm exposed trout was most pronounced when rates at 15 degrees C were expressed relative to cytochrome a and c(1) levels. Non-phosphorylating (State 4) rates of oxygen uptake increased with short-term warm exposure before returning to initial levels after warm acclimation. Cytochrome c oxidase (CCO) activity in the mitochondrial preparations decreased with warm acclimation. The thermal sensitivity of the ADP affinity was markedly modified during short-term warm exposure, when the ADP/O ratio increased, but warm acclimation returned these values to those observed initially. ADP affinity increased after warm acclimation. Changes in the mitochondrial content of cytochromes and adenine nucleotide translocase (ANT) could not explain these patterns. On the other hand, changes in the proportions of the lipid classes and in the acyl chain composition of certain phospholipid classes mirror the modifications in functional properties. Short-term exposure to 15 degrees C decreased the ratio of diacylphosphatidylethanolamine/diacylphosphatidylcholine (diacylPE/diacylPC), whereas warm acclimation led to restructuring of fatty acids (FA) and to increases of plasmalogen forms of PE and PC. Modification of overall membrane unsaturation did not appear to be the primary aim of restructuring membrane FA during warm acclimation, as total mitochondrial phospholipids and the major phospholipid classes only showed slight shifts of their acyl composition with warm acclimation. On the other hand, natural lysophosphatidylcholine (LysoPC) showed dramatic changes in FA content, as 16:0 and 18:1n-9 doubled whereas 22:6n-3 decreased from around 50% to 32% in warm acclimated trout. Similarly, in cardiolipin (CL), the levels of 16:0 and 18:1n-7 halved while 18:2n-6 increased to over 20% of the FA with warm acclimation. Given the central role of CL in modulating the activity of CCO, F(0)F(1)-ATPase and ANT, these changes suggest that specific compositional changes in CL are important modulators of mitochondrial capacities. The many structural changes in membrane lipids contrast with the limited modifications of the membrane protein components examined and support the concept of lipid structure modulating mitochondrial capacities.

Acclimatization↗

The effects of temperature acclimation on the resting membrane of skeletal muscle fibres from green sunfish.

Conductive properties of muscle fibres from green sunfish (Lepomis cyanellus) acclimated to different temperatures were examined. The relative membrane permeability to chloride and potassium ions, PCl/PK, measured at acclimation temperature, was approximately 7.0 after acclimation at 25 degrees C and 1.3 after acclimation at 7 degrees C. This difference was due to a six-fold reduction in the membrane chloride conductance upon acclimation to 7 degrees C as compared to 25 degrees C-acclimated fibres. Mean (+/- S.E.M.) values of the chloride conductance were 554 +/- 68 microseconds cm-2 in warm-acclimated sunfish, and 75 +/- 9 microseconds cm-2 in cold-acclimated sunfish. Membrane capacitance also differed significantly between the two acclimation groups. When the temperature was varied acutely, the magnitude of the chloride conductance exhibited a maximum Q10 of only 1.9, compared with a Q10 of 3.0 associated with acclimation. Upon transferring 25 degrees C-acclimated sunfish to holding tanks at 7 degrees C, the total membrane resistance exhibited a sigmoidal increase over about 14 days, and a steady membrane capacitance was achieved in about 10 days. For 7 degrees C-acclimated sunfish, transferred to 25 degrees C, resistance showed a sigmoidal decrease over 10 days and capacitance was steady after 8 days. The results indicate that thermal acclimation of the muscle membrane involves cellular regulatory processes which underlie significant changes in the electrical properties of the fibre.

Animals↗

Molecular mechanisms underlying the plasticity of muscle contractile properties with temperature acclimation in the marine fish Myoxocephalus scorpius

Single fibres were isolated from the fast myotomal muscle of the short-horned sculpin (Myoxocephalus scorpius L.). Fish were acclimated to either 5 &deg;C (10 h:14 h light:dark) or 15 &deg;C (12 h:12 h light:dark) for 1&shy;2 months. Isometric tension (Fmax) and unloaded contraction velocity (Vslack) were determined in maximally activated skinned fibres over the range 0 to 20 &deg;C. Fibres isolated from 5 &deg;C-acclimated and 15 &deg;C-acclimated fish failed to relax completely following activations at 15 &deg;C and 20 &deg;C respectively. In 5 &deg;C-acclimated fish, Fmax increased from 75 kN m-2 at 0 &deg;C to 123 kN m-2 at 10 &deg;C and was not significantly higher at 15 or 20 &deg;C. The relationship between Fmax and temperature was not significantly different for cold- and warm-acclimated fish. Vslack was around 2.8 fibre lengths s-1 in both acclimation groups at 0 &deg;C, but it increased at a significantly faster rate with temperature in 15 &deg;C- than in 5 &deg;C-acclimated fish. At 20 &deg;C, Vmax was significantly higher in 15 &deg;C-acclimated (8.7 fibre lengths s-1) than in 5 &deg;C-acclimated fish (5.3 fibre lengths s-1). In order to investigate the molecular mechanism(s) underlying changes in Vmax, myosin was purified by ion-exchange chromatography. No difference in myosin heavy chain composition could be detected on the basis of peptide mapping with four different proteolytic enzymes. Two-dimensional polyacrylamide gel electrophoresis revealed no myofibrillar protein isoforms unique to either acclimation temperature. However, the ratio of myosin alkali light chain contents (LC3f:LC1f), as determined by capillary electrophoresis, was significantly lower in muscle from 15 &deg;C-acclimated (0.73) than from 5 &deg;C-acclimated fish (1.66). The results suggest that changes in Vmax are achieved via altered expression of myosin light chains independently of myosin heavy chain composition. In support of this hypothesis, the myofibrillar ATPase activity of fast muscle was not altered by temperature acclimation.

Journal Article↗

The influence of thermal acclimation on power production during swimming. I. In vivo stimulation and length change pattern of scup red muscle.

Ectothermal animals are able to locomote in a kinematically similar manner over a wide range of temperatures. It has long been recognized that there can be a significant reduction in the power output of muscle during swimming at low temperatures because of the reduced steady-state (i.e. constant activation and shortening velocity) power-generating capabilities of muscle. However, an additional reduction in power involves the interplay between the non-steady-state contractile properties of the muscles (i.e. the rates of activation and relaxation) and the in vivo stimulation and length change pattern the muscle undergoes during locomotion. In particular, it has been found that isolated scup (Stenotomus chrysops) red muscle working under in vivo stimulus and length change conditions (measured in warm-acclimated scup swimming at low temperatures) generates very little power for swimming. Even though the relaxation of the muscle has slowed greatly, warm-acclimated fish swim with the same tail-beat frequencies and the same stimulus duty cycles at cold temperatures, thereby not affording the slow-relaxing muscle any extra time to relax. We hypothesize that considerable improvement in the power output of the red muscle at low temperatures could be achieved if cold acclimation resulted in either a faster muscle relaxation rate or in the muscle being given more time to relax (e.g. by shortening the stimulus duration or reducing the tail-beat frequency). We test these hypotheses in this paper and the accompanying paper. Scup were acclimated to 10 degrees C (cold-acclimated) and 20 degrees C (warm-acclimated) for at least 6 weeks. Electromyograms (EMGs) and high-speed cine films were taken of fish swimming steadily at 10 degrees C and 20 degrees C. At 10 degrees C, we found that, although there were no differences in tail-beat frequency, muscle strain or stimulation phase between acclimation groups, cold-acclimated scup had EMG duty cycles approximately 20 % shorter than warm-acclimated scup. In contrast at 20 degrees C, there was no difference between acclimation groups in EMG duty cycle, nor in any other muscle length change or stimulation parameter. Thus, in response to cold acclimation, there appears to be a reduction in EMG duty cycle at low swimming temperatures that is probably due to an alteration in the operation of the pattern generator. This novel acclimation probably improves muscle power output at low temperatures compared with that of warm-acclimated fish, an expectation we test in the accompanying paper using the work-loop technique.

Adaptation, Physiological↗

Interplay among energy metabolism, organ mass and digestive enzyme activity in the mouse-opossum Thylamys elegans: the role of thermal acclimation.

The potential for thermal acclimation in marsupials is controversial. Initial studies suggest that the thermoregulatory maximum metabolic rate (MMR) in metatherians cannot be changed by thermal acclimation. Nevertheless, recent studies reported conspicuous seasonality in both MMR and in basal metabolic rate (BMR). We studied the role of thermal acclimation in the Chilean mouse-opossum, Thylamys elegans, by measuring MMR and BMR before and after acclimation to cold or warm conditions. Following acclimation we also measured the mass of metabolically active organs, and the activity of a key digestive enzyme, aminopeptidase-N. No significant effect of thermal acclimation (i.e. between cold- and warm-acclimated animals) was observed for body mass, MMR, body temperature or factorial aerobic scope. However, the BMR of cold-acclimated animals was 30 % higher than for warm-acclimated individuals. For organ mass, acclimation had a significant effect on the dry mass of caecum, liver and kidneys only. Stepwise multiple regression using pooled data showed that 71 % of the variation in BMR is explained by the digestive organs. Overall, these results suggest that MMR is a rather rigid variable, while BMR shows plasticity. It seems that T. elegans cannot respond to thermal acclimation by adjusting its processes of energy expenditure (i.e. thermogenic capacity and mass of metabolically active organs). The lack of any significant difference in aminopeptidase-N specific activity between warm- and cold-acclimated animals suggests that this response is mainly quantitative (i.e. cell proliferation) rather than qualitative (i.e. differential enzyme expression). Finally, as far as we know, this study is the first to report the effects of thermal acclimation on energy metabolism, organ mass and digestive enzyme activity in a marsupial.

Acclimatization↗

Calorigenic effects of noradrenaline and glucagon on white adipocytes in cold- and heat-acclimated rats.

Calorigenic actions of noradrenaline and glucagon on isolated epididymal fat cells from warm-acclimated controls, cold-acclimated and heat-acclimated rats were measured by the use of a twin-type conduction microcalorimeter. Both noradrenaline and glucagon stimulated heat production in isolated adipocytes maximally in doses of 1 microgram/ml and 10 microgram/ml, respectively. Maximal responsiveness of adipocytes per unit cell to noradrenaline was not influenced by cold acclimation, while it was reduced by heat and acclimation. Maximal response in total epididymal fat cells to noradrenaline was increased in cold acclimation and not changed in heat acclimation at increased numbers of adipocytes in both cold-acclimated and heat-acclimated animals. Maximal response per unit cell as well as per total epididymal fat cells to glucagon was increased in cold acclimation and reduced in heat acclimation. The present results indicate that the modified responses of target adipocytes to noradrenaline and glucagon are involved in the development of temperature acclimation.

Adaptation, Physiological↗

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

The acclimation temperature of carp does not affect the amount of cytochrome c oxidase per mg mitochondrial protein as revealed from the reduced-minus-oxidized difference spectra of red muscle mitochondria from cold- and warm-acclimated carp. There are no differences between cold- and warm-acclimated fish in the substrate binding properties of the enzyme as judged from the Km values for cytochrome c at 30 degrees C (3.34 +/- 0.ee microM, acclimation temperature 10 degrees C and 3.55 +/- 0.31 microM, acclimation temperature 30 degrees C). The molar activities of the enzyme, however, differ for both acclimation temperatures: when intercalated in the 10 degrees C-acclimated mitochondrial membrane, the enzyme can catalyze the oxidation of 117.6 +/- 17.2 mol ferrocytochrome c/s per mol heme a as compared with 85.6 +/- 17.2 in the 30 degrees C-acclimated membrane (experimental temperature 30 degrees C). Correspondingly, higher specific activities of the succinate oxidase system are observed in mitochondria from cold-acclimated carp as compared with those obtained from warm-acclimated carp. The results indicate that cold acclimation of the eurythermic carp is accompanied by a partial compensation of the acute effect of decreasing temperature on the activity of cytochrome c oxidase in red muscle mitochondria. Based on the temperature-induced lipid adaptation reported for carp red muscle mitochondria (Wodtke, E. (1980) Biochim. Biophys. Acta 640, 698--709), it is concluded that during thermal acclimation the molar activity of cytochrome c oxidase is controlled by viscotropic regulation. The results fit to the conception that cardiolipin constitutes a lipid shell (annulus) surrounding the oxidase within the native membrane, but that it is the bilayer fluidity and not the annular fluidity which determines the activity of cytochrome c oxidase.

Acclimatization↗

Multi-generation cadmium acclimation and tolerance in Daphnia magna Straus.

The cladoceran Daphnia magna was acclimated for seven generations to cadmium concentrations ranging from 0 (control) to 250 microg/l Cd (corresponding to a free ion activity of 4.60 nM Cd2+). Acute and chronic cadmium tolerance as well as cadmium accumulation were monitored as a function of acclimation time. After two to three generations of acclimation to concentrations ranging from 0.23 to 1.11 nM Cd2+ increases in acute tolerance were maximal (factor 7.2) and significant. Acclimation for seven generations to the same acclimation concentrations did result in an increased chronic cadmium tolerance (21 days EC50 values increased). Organisms acclimated to 1.93 nM Cd2+ were equally or more sensitive than non-acclimated daphnids in acute and chronic toxicity tests. Cadmium contents in D. magna increased significantly as a function of the acclimation concentration. Maximum body burdens of 236+/-30 microg Cd/g dry weight were measured in organisms exposed to 4.60 nM Cd2+, but detoxification mechanisms were only successful up to 82+/-20 microg Cd/g dry weight as this concentration did not cause major decreases in survival and reproduction in chronic toxicity tests. As the potential positive effect of acclimation on cadmium tolerance disappeared with successive acclimation generations and increasing acclimation concentrations, it is concluded that multi-generation acclimation studies are important for the evaluation of the long-term effects of environmental toxicants.

Acclimatization↗

Daily photosynthetic and C-export patterns in winter wheat leaves during cold stress and acclimation.

Diurnal patterns of whole-plant and leaf gas exchange and 14C-export of winter wheat acclimated at 20 and 5 degrees C were determined. The 5 degrees C-acclimated plants had lower relative growth rates, smaller biomass and leaf area, but larger specific leaf weight than 20 degrees C plants. Photosynthetic rates in 20 degrees C and 5 degrees C-acclimated leaves were similar; however, daytime export from 5 degrees C-acclimated leaves was 45% lower. Photosynthesis and export remained steady in 20 degrees C and 5 degrees C-acclimated leaves during the daytime. By comparison, photosynthesis in 5 degrees C-stressed leaves (20 degrees C-acclimated plants exposed to 5 degrees C 12 h before and during measurements) declined from 70 to 50% of the 20 degrees C-acclimated leaves during the daytime, while export remained constant at 35% of the 20 degrees C-acclimated and 60% of the 5 degrees C-acclimated leaves. At high light and CO2, photosynthesis and export increased in both 20 degrees C and 5 degrees C-acclimated leaves, but rates in 5 degrees C-stressed leaves remained unchanged. At all conditions daytime export was greater than nighttime export. Taken together, during cold acclimation photosynthesis was upregulated, whereas export was only partially increased. We suggest that this reflects a requirement of cold-acclimated plants to both sustain an increased leaf metabolic demand while concomitantly supporting translocation of photoassimilates to overwintering sinks.

Journal Article↗

Effects of atmospheric humidity and acclimation temperature on the temperature response of photosynthesis in young Larix decidua Mill.

Larch (Larix decidua Mill.) seedlings of a low altitude (600 m) Austrian provenance were raised outdoors and acclimated in chambers for 14 to 24 days during August and September at either 8 degrees C and an atmospheric saturation vapor pressure deficit (DeltaW) of 2.5 Pa kPa(-1), or 24 degrees C and a DeltaW of 6.2 Pa kPa(-1). Subsequently, their rates of photosynthesis, dark respiration and transpiration were measured at temperatures between 5 and 30 degrees C with DeltaW either maintained below 10 Pa kPa(-1) or allowed to increase with temperature up to 38 Pa kPa(-1). Below 15 degrees C the photosynthetic rate of cold-acclimated plants was higher, but above 15 degrees C it was lower, than that of warm-acclimated plants. Temperature acclimation caused a greater shift in the temperature optimum for photosynthesis when DeltaW was kept small than when it was allowed to increase with temperature. When DeltaW was kept small, leaf conductance of cold-acclimated plants, unlike that of warm-acclimated plants, did not increase with temperature above 15 degrees C. When DeltaW increased with temperature, leaf conductance of cold-acclimated plants decreased more rapidly with temperature than that of warm-acclimated plants. Low temperature acclimation increased the rate of photosynthesis below 15 degrees C without affecting leaf conductance, which indicates that there was an adaptation in leaf internal processes. Further evidence of a metabolic adaptation to acclimation temperature is that dark respiration of cold-acclimated plants was twice that of warm-acclimated plants at all temperatures.

Journal Article↗

Cold acclimation of Arabidopsis thaliana results in incomplete recovery of photosynthetic capacity, associated with an increased reduction of the chloroplast stroma.

The effects of short-term cold stress and long-term cold acclimation on the light reactions of photosynthesis were examined in vivo to assess their contributions to photosynthetic acclimation to low temperature in Arabidopsis thaliana (L.) Heynh.. All photosynthetic measurements were made at the temperature of exposure: 23 degrees C for non-acclimated plants and 5 degrees C for cold-stressed and cold-acclimated plants. Three-day cold-stress treatments at 5 degrees C inhibited light-saturated rates of CO2 assimilation and O2 evolution by approximately 75%. The 3-day exposure to 5 degrees C also increased the proportion of reduced QA by 50%, decreased the yield of PSII electron transport by 65% and decreased PSI activity by 31%. In contrast, long-term cold acclimation resulted in a strong but incomplete recovery of light-saturated photosynthesis at 5 degrees C. The rates of light-saturated CO2 and O2 gas exchange and the in vivo yield of PSII activity under light-saturating conditions were only 35-40% lower, and the relative redox state of QA only 20% lower, at 5 degrees C after cold acclimation than in controls at 23 degrees C. PSI activity showed full recovery during long-term cold acclimation. Neither short-term cold stress nor long-term cold acclimation of Arabidopsis was associated with a limitation in ATP, and both treatments resulted in an increase in the ATP/NADPH ratio. This increase in ATP/NADPH was associated with an inhibition of PSI cyclic electron transport but there was no apparent change in the Mehler reaction activity in either cold-stressed or cold-acclimated leaves. Cold acclimation also resulted in an increase in the reduction state of the stroma, as indicated by an increased total activity and activation state of NADP-dependent malate dehydrogenase, and increased light-dependent activities of the major regulatory enzymes of the oxidative pentose-phosphate pathway. We suggest that the photosynthetic capacity during cold stress as well as cold acclimation is altered by limitations at the level of consumption of reducing power in carbon metabolism.

Acclimatization↗

Exercise- and methylcholine-induced sweating responses in older and younger men: effect of heat acclimation and aerobic fitness.

The purpose of this investigation was to examine the effects of aging and aerobic fitness on exercise- and methylcholine-induced sweating responses during heat acclimation. Five younger [Y group-age: 23 +/- 1 (SEM) years; maximal oxygen consumption (VO2max): 47 +/- 3 ml.kg-1.min-1], four highly fit older (HO group- 63 +/- 3 years; 48 +/- 4 ml.kg-1.min-1) and five normally fit older men (NO group -67 +/- 3 years; 30 +/- 1 ml.kg-1.min-1) who were matched for height, body mass and percentage fat, were heat acclimated by daily cycle exercise (approximately 35% VO2max for 90 min) in a hot (43 degrees C, 30% RH) environment for 8 days. The heat acclimation regimen increased performance time, lowered final rectal temperature (Tre) and percentage maximal heart rate (%HRmax), improved thermal comfort and decreased sweat sodium concentration similarly in all groups. Although total body sweating rates (Msw) during acclimation were significantly greater in the Y and HO groups than in the NO group (P < 0.01) (because of the lower absolute workload in the NO group), the Msw did not change in all groups with the acclimation sessions. Neither were local sweating rates (msw) on chest, back, forearm and thigh changed in all groups by the acclimation. The HO group presented greater forearm msw (30-90 min) values and the Y group had greater back and thigh msw (early in exercise) values, compared to the other groups (P < 0.001). In a methylcholine injection test on days immediately before and after the acclimation, the order of sweat output per gland (SGO) on chest, back and thigh was Y > HO > NO, and on the forearm Y = HO > NO. No group differences were observed for activated sweat gland density at any site. The SGO at the respective sites increased in the post-acclimation test regardless of group (P < 0.01), but on the thigh the magnitude of the increase was lower in the NO (P < 0.02) and HO (P = 0.07) groups than in the Y group. These findings suggest that heat tolerance and the improvement with acclimation are little impaired not only in highly fit older but also normally fit older men, when the subjects exercised at the same relative exercise intensity. Furthermore, the changes induced by acclimation appear associated with an age-related decrease in VO2max. However methylcholine-activated SGO and the magnitude of improvement of SGO with acclimation are related not only to VO2max but also to aging, suggesting that sensitivity to cholinergic stimulation decreases with aging.

Adaptation, Physiological↗

Effects of thermal acclimation on nervous conduction and muscle contraction in the frog Rana temporaria.

The effects of season and acclimation temperature on the latency of the leg withdrawal reflex and three of its components have been studied: conduction velocity in the sciatic nerve, spinal conduction time, and contraction time of gastrocnemius muscle. The latency of the leg withdrawal reflex was markedly shortened by cold acclimation: the reaction times were at 6 degrees C 1.54 s in 4 degrees C acclimated and 3.97 s in 24 degrees C acclimated winter frogs. Also, the temperature dependence of the reflex latency was reduced by cold acclimation. Thus, frogs acclimated to cold responded to external stimuli in cold more rapidly than warm-acclimated ones. This cold adaptation of the reflex could not be explained by changes in its studied components. These made up only one-tenth of the reflex response time, and either did not show significant cold acclimation (muscle contraction and spinal conduction times in summer) or showed inverse acclimation, especially when measured at high temperatures (i.e. conduction velocities were reduced by acclimation to cold). Thus, the cold acclimation of the reflex response probably resides in the sensory component of the response. The inverse temperature adaptation response of conduction velocities may reflect a reduced ion permeability across cellular membranes in cold which decreases metabolic energy expenditure during inactive periods.

Adaptation, Physiological↗

Phylogenetic analysis of thermal acclimation of the glycolytic enzymes in the genus Fundulus.

Physiological acclimation that alters enzyme activity can compensate for the effect of temperature on function and may be achieved by altering enzyme concentration. This study uses phylogenetic analyses to investigate the evolutionary history of and to test several hypotheses about acclimation responses among all the glycolytic enzymes. These hypotheses are that (1) acclimation increases enzyme concentration at lower temperatures to compensate for reduced activity; (2) equilibrium enzymes tend to show acclimation responses; and (3) acclimation responses are more common in species whose populations experience either large temporal or geographical temperature variations. Using maximal activities as indices of enzyme concentration, the presence of acclimation responses in all the glycolytic enzymes in the heart ventricle was determined for five species in the teleost genus Fundulus. Three of these species are distributed along the steep thermal cline of the North American Atlantic coast, and thus these species experience both seasonal and geographical variation in temperature. The other two species are found in the Gulf of Mexico and experience seasonal variation similar to the Atlantic species but no geographical variation in temperature. Two Atlantic coast species, Fundulus heteroclitus and Fundulus majalis, have unique derived acclimation responses. No derived acclimation responses occur in the Gulf species. A conserved response in hexokinase was observed within one subgenus comprising both Atlantic and Gulf species. In F. heteroclitus, enolase responded to acclimation, and in F majalis, aldolase, triphosphate isomerase, and lactate dehydrogenase had acclimation responses. These enzymes are equilibrium enzymes, and the concentrations of all of them increase at lower temperatures, which would compensate for the effect of temperature on enzyme activity. The compensatory changes all occur in the Atlantic species and may be a mechanism for species to expand their ranges. These data suggest that physiological acclimation is evolutionarily labile.

Acclimatization↗

Induction of freezing tolerance in spinach is associated with the synthesis of cold acclimation induced proteins.

Spinach (Spinacia oleracea L. cv Bloomsdale) seedlings cultured in vitro were used to study changes in protein synthesis during cold acclimation. Seedlings grown for 3 weeks postsowing on an inorganic-nutrient-agar medium were able to increase their freezing tolerance when grown at 5 degrees C. During cold acclimation at 5 degrees C and deacclimation at 25 degrees C, the kinetics of freezing tolerance induction and loss were similar to that of soil-grown plants. Freezing tolerance increased after 1 day of cold acclimation and reached a maximum within 7 days. Upon deacclimation at 25 degrees C, freezing tolerance declined within 1 day and was largely lost by the 7th day. Leaf proteins of intact plants grown at 5 and 25 degrees C were in vivo radiolabeled, without wounding or injury, to high specific activities with [(35)S]methionine. Leaf proteins were radiolabeled at 0, 1, 2, 3, 4, 7, and 14 days of cold acclimation and at 1, 3, and 7 days of deacclimation. Up to 500 labeled proteins were separated by two-dimensional gel electrophoresis and visualized by fluorography. A rapid and stable change in the protein synthesis pattern was observed when seedlings were transferred to the low temperature environment. Cold-acclimated leaves contained 22 polypeptides not found in nonacclimated leaves. Exposure to 5 degrees C induced the synthesis of three high molecular weight cold acclimation proteins (CAPs) (M(r) of about 160,000, 117,000, and 85,000) and greatly increased the synthesis of a fourth high molecular weight protein (M(r) 79,000). These proteins were synthesized during day 1 and throughout the 14 day exposure to 5 degrees C. During deacclimation, the synthesis of CAPs 160, 117, and 85 was greatly reduced by the first day of exposure to 25 degrees C. However, CAP 79 was synthesized throughout the 7 day deacclimation treatment. Thus, the induction at low temperature and termination at warm temperature of the synthesis of CAPs 160, 117, and 85 was highly correlated with the induction and loss of freezing tolerance. Cold acclimation did not result in a general posttranslational modification of leaf proteins. Most of the observed changes in the two-dimensional gel patterns could be attributed to the de novo synthesis of proteins induced by low temperature. In spinach leaf tissue, heat shock altered the pattern of protein synthesis and induced the synthesis of several heat shock proteins (HSPs). One polypeptide synthesized in cold-acclimated leaves had a molecular weight and net charge (M(r) 79,000, pI 4.8) similar to that of a HSP (M(r) 83,000, pI 4.8). However, heat shock did not increase the freezing tolerance, and cold acclimation did not increase heat tolerance over that of nonacclimated plants, but heat-shocked leaf tissue was more tolerant to high temperatures than nonacclimated or cold-acclimated leaf tissue. When protein extracts from heat-shocked and cold-acclimated leaves were mixed and separated in the same two-dimensional gel, the CAP and HSP were shown to be two separate polypeptides with slightly different isoelectric points and molecular weights.

Journal Article↗