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Brown adipose tissue thermogenesis is activated by electrical and chemical (L-glutamate) stimulation of the ventromedial hypothalamic nucleus in cold-acclimated rats.

Experiments were conducted to determine if both electrical and chemical stimulation of the ventromedial hypothalamic nucleus (VMH) could activate brown adipose tissue (BAT) thermogenesis. Age-matched, room-acclimated (21 degrees C) and cold-acclimated (4 degrees C for 3 weeks prior to testing) male Sprague-Dawley rats were given unilateral electrical or chemical stimulation to the VMH by way of a 'chemotrode apparatus'. The devised 'chemotrode' allowed both electrical stimulation (insulated piano wire stimulating electrode) and chemical stimulation (23 gauge stainless steel intracranial cannula of equal length) to be performed at the same VMH site using a common 19 gauge stainless steel outer guide tube. The first unilateral VMH electrical stimulation (0.5 ms pulse, 50 Hz and 120 microA for 30 s) caused no significant rise in interscapular brown adipose tissue temperature (TIBAT) colonic (Tc) or tail surface temperatures (Tt), compared to respective prestimulation control values in rats acclimated to 21 degrees C. In the 4 degrees C-acclimated group the first VMH electrical stimulation caused a significant rise in IBAT temperature. L-Glutamate administration to the same VMH site (60 nmol in 600 nl volume) also caused a significant increase in IBAT temperature in the 4 degrees C but not the 21 degrees C-acclimated rats. The rise in IBAT temperature following the L-glutamate injection to the 4 degrees C-acclimated group was similar to that found following the first electrical stimulation to this group. Interestingly, a second unilateral electrical stimulation of the VMH to 4 degrees C-acclimated rats could not evoke a similar increase in IBAT temperature suggesting that overall L-glutamate was acting in vivo as an excitotoxin.(ABSTRACT TRUNCATED AT 250 WORDS)

Acclimatization↗

Streptozotocin diabetes in rats after acclimation to cold environment.

To investigate the effects of acclimation to a cold environment on the alteration of glucose and fat metabolism in diabetes, the levels of blood glucose, triacylglycerol, nonesterified fatty acid, and immunoreactive insulin were measured before and on the 3rd and 14th days after streptozotocin treatment in cold-acclimated (4 degrees C) and warm-acclimated (24 degrees C; control) rats. The cold-acclimated rats showed no hyperglycemia on the 14th day after streptozotocin treatment, whereas the control diabetic rats maintained high blood glucose levels. Triacylglycerol levels did not increase in the cold-acclimated diabetic rats, whereas the control diabetic rats showed markedly high triacylglycerol levels. Immunoreactive insulin levels remained constantly low in both cold-acclimated and control diabetic rats. It was concluded that acclimation to cold changed some of the metabolic consequences of diabetes caused by streptozotocin in rats.

Acclimatization↗

Enzymatic changes in mitochondria-rich cells of Xenopus laevis skin epithelium are induced by ionic acclimation.

Morphological, biochemical and histochemical components of mitochondria-rich (MR) cells of skin epithelium of Xenopus laevis (Daudin) were investigated after acclimation in distilled water (DW) and mild solutions (50 mmol/l) of either NaCl or KCl for over 10 days. The animals readily acclimated to NaCl, but approximately 50% of the animals died in the KCl solution. Electrophysiological measurements confirmed the poor transport properties of skin in all conditions. Silver staining and exposure to methylene blue (MB) have shown that two types of MR cells can be distinguished, especially after KCl acclimation. Immunohistochemistry with the use of anti-band 3 polyclonal and anti H+-ATPase monoclonal antibodies demonstrated that band 3 and H+-ATPase enzymes were localized in MR cells in all conditions. H+-ATPase was greatly reduced during NaCl acclimation as verified with SDS gel electrophoresis. Intensity of the immunohistochemical staining differed between the various conditions of acclimation. Histochemical localization of carbonic anhydrase and alkaline phosphatase activities was more intense during NaCl acclimation. Morphological changes were also observed between the various acclimation conditions. The present findings substantiate the existence of at least two forms of MR cells in Xenopus skin epithelium but their functional significance remains to be established.

Adaptation, Physiological↗

Membrane order conservation in raft and non-raft regions of hepatocyte plasma membranes from thermally acclimated rainbow trout.

Homeoviscous adaptation (HVA), the thermal conservation of membrane fluidity/order at different body temperatures, has been observed to varying degrees in different membranes. However, HVA has not been studied in raft and non-raft regions of the plasma membrane (PM) separately. Rafts are ordered PM microdomains implicated in signal transduction, membrane traffic and cholesterol homeostasis. Using infrared spectroscopy, we measured order in raft-enriched PM (raft) and raft-depleted PM (RDPM) isolated from hepatocytes of rainbow trout (Oncorhynchus mykiss) acclimated to 5 and 20 degrees C. We found approximately 130% and 90% order compensation in raft and RDPM, respectively, suggesting their independent regulation. Raft was more ordered than RDPM in the warm-acclimated trout, a difference fully explained by a 58% enrichment of cholesterol, compared to RPDM. Unexpectedly, raft and RDPM from cold-acclimated trout did not differ in cholesterol content or order. Freezing the membrane samples during preparation had no effect on order. Treatment with cyclodextrin depleted cholesterol by 36%, 56%, and 55%, producing significant decreases in order in raft and RDPM from warm-acclimated trout and RDPM from cold-acclimated trout, respectively. However, a 69% depletion of cholesterol from raft from cold-acclimated trout had no significant effect on order. This result, and the lack of a difference in order between raft and RDPM, suggests that raft and non-raft PM in cold-acclimated trout are not spatially segregated by phase separation due to cholesterol.

Animals↗

Characterization of ion channel and transporter mRNA expressions in isolated gill chloride and pavement cells of seawater acclimating eels.

Ion channels and transporters (i.e. cystic fibrosis transmembrane regulator (CFTR), inward rectifier potassium channel (eKir), Na/K-ATPase, Na/K/Cl2 co-transporter (NKCC), aquaporin-3 (AQP-3), and Na/H exchanger-1 (NHE-1)) are known to be expressed in gill epithelia of teleost fish. Owing to the anatomical complexity of gill structures, their temporal expression profile in seawater acclimating gill pavement (PVCs) and chloride cells (CCs) are limited. In this study, we isolated the gill PVCs and CCs from seawater acclimating Japanese eels to address the issue. In the gill epithelia of freshwater adapted eels, CCs expressed the highest mRNA and/or protein levels of Na/K-ATPase, NKCC, and eKir as demonstrated by real-time PCR and/or immunohistochemical staining. AQP-3 mRNA was highly expressed in freshwater PVCs and its protein was in general expressed in all gill cells. The NHE-1 transcripts were expressed in similar levels in both PVCs and CCs. CFTR mRNA transcript was almost undetectable in all the freshwater gill cell samples. Seawater acclimation induced the transcript and/or protein levels of Na/K-ATPase, NKCC, CFTR, and eKir in CCs. The upregulation and the coexpression of these transporters in CCs suggested their cohort function in mediating Na+, K+, and Cl- transport. The expression of CFTR was found to be tightly regulated as its expression was restricted only in "seawater CCs". AQP-3 transcript and protein levels in PVCs reduced significantly during the acclimation. Interestingly immunocytochemical (ICC) staining of seawater gill epithelia revealed that AQP-3 immunoreactivities were mainly localized in seawater CCs. In the acclimation, there was no significant reduction of NHE-1 mRNA in both PVCs and CCs, however its protein level dropped significantly in the seawater condition. The present study is the first to demonstrate the activation of the mRNA transcripts for the ion channels and transporters in isolated gill CCs during seawater acclimation. The activating mechanism is found to be confined primarily in CCs. These results indicated that in addition to the increase in size and number of CCs, the molecular remodeling and the functional plasticity of CCs were essential in the ion transport process during seawater acclimation.

Acclimatization↗

The effect of temperature acclimation on myocardial beta-adrenoceptor density and ligand binding affinity in African catfish (Claris gariepinus).

This study assessed the effects of temperature acclimation on myocardial beta-adrenoceptor density (B(max)) and binding affinity (K(d)) in African catfish (Claris gariepinus) acclimated to 15, 22 and 32 degrees C. B(max) values were not significantly different (P > 0.05) among the three acclimation groups. Conversely, the K(d) value of the 32 degrees C acclimation group (K(d) = 0.88) was significantly higher (P = 0.002) than both the 15 degrees C (K(d) = 0.48) and 22 degrees C (K(d) = 0.46) acclimation groups. In addition, K(d) of rainbow trout (Oncorhynchus mykiss) was significantly lower (P < 0.001) and B(max) significantly higher (P < 0.05) than that of African catfish at all three acclimation temperatures. These results contrast with those reported previously for temperate species, in which B(max) is inversely related to acclimation temperature, and counter a previous suggestion that B(max) is higher in tropical versus temperate species.

Acclimatization↗

Multigeneration acclimation of Daphnia magna Straus to different bioavailable copper concentrations.

A large acclimation experiment was performed with Daphnia magna in which two different copper bioavailability (as Cu2+) groups (N and M) were used. In the N group the cupric ion activity increased with increasing dissolved copper-acclimation concentration, while in the M group the ion activity decreased with increasing dissolved copper concentration. The activity of copper carbonates and hydroxides was up to an order of magnitude lower than the cupric ion activity. After five generations of acclimation, the acute copper sensitivity (mean +/- SD) of the N group ranged from 193 /- 24 to 296 +/- 50 microg Cu L(-1) and for the M group from 198 +/- 27 to 315 +/- 38 microg Cu L(-1) for daphnids acclimated to 1 and 100 microg Cu L(-1), respectively. The internal copper concentration of the acclimated daphnids also resulted in similar results between the two groups. Acclimation of the two daphnid groups for five consecutive generations to the three dissolved copper concentrations resulted in a shift in the optimal concentration range toward 1 microg Cu L(-1), using energy reserves as an endpoint. Our results suggest that copper acclimation and accumulation are related to the dissolved copper concentration of the culture medium, but not to the copper activity.

Adaptation, Physiological↗

Plasma membrane lipid alterations induced by cold acclimation and abscisic acid treatment of winter wheat seedlings differing in frost resistance.

Cold acclimation of plants affects many aspects of metabolism. Changes in plasma membrane lipids have always been considered to be important for development of frost resistance and survival at subzero temperatures. We studied different cultivars of winter wheat (Triticum aestivum L.) that differed in frost resistance induced either by cold acclimation or treatment with the plant hormone abscisic acid (ABA). Plasma membranes were isolated from non-acclimated and cold- as well as from ABA-acclimated plants, and were subjected to detailed lipid analysis. Cold acclimation affected virtually all plasma membrane lipid components and their constituents, resulting in both increases and decreases, which varied between the three groups of plants investigated. Including the cold-induced variations observed in the few plant species studied in detail previously, cerebrosides were the only components reduced by cold acclimation in all plants. In wheat, more uniform and consistent patterns were obtained when considering colligative parameters such as total free sterols, phospholipids or glycolipids, either as the proportion of total lipids or based on plasma membrane protein. The parameter which changed most significantly in parallel to the increase of inducible frost resistance in the three groups of plants was the ratio of free sterols/glycolipids, which increased. ABA treatment resulted in qualitatively similar effects in only one cultivar, but in general these changes were less pronounced. Compared to changes in transcription rates of several cold-induced genes and in the concentration of various compatible solutes reported for other plants, the observed changes in plasma membrane lipids are minor ones. This may indicate that acclimation-induced changes can be accomplished by posttranscriptional regulation of enzymatic activities, which is in agreement with the failure to detect significant changes in transcription of the corresponding genes during cold induction.

Abscisic Acid↗

Heat acclimation prolongs the time to central nervous system oxygen toxicity in the rat. Possible involvement of HSP72.

Oxygen toxicity of the central nervous system (CNS-OT) can occur during diving with oxygen-enriched gas mixtures, or during hyperbaric medical treatment. CNS-OT is characterised by convulsions and sudden loss of consciousness, which may be fatal in diving. Heat acclimation is known to provide cross-tolerance to various forms of stress in different organs, including the brain. We hypothesised that heat acclimation may delay the onset of CNS-OT in the rat. Male Sprague-Dawley rats were acclimated to an ambient temperature of 32 degrees C for 4 weeks. Rats in the control group were kept at 24 degrees C. Both groups were exposed to oxygen at 608 kPa. EEG was recorded continuously until the appearance of the first electrical discharge preceding clinical convulsions. CO(2) production was measured simultaneously with the EEG. Latency to CNS-OT was measured and brain samples were taken for evaluation of heat shock protein 72 (HSP72) levels by Western blot analysis at the end of the acclimation period and during 4 weeks of deacclimation. Latency to CNS-OT was twice as long in the heat-acclimated rat, with insignificant changes in CO(2) production. This prolongation continued for 2 weeks during deacclimation. There was a significant increase in the level of HSP72 following heat acclimation, with a subsequent decrease during deacclimation. We conclude that heat acclimation prolongs latency to CNS-OT in a way that does not involve changes in metabolic rate. During deacclimation there was a linear relationship between latency to CNS oxygen toxicity and the level of HSP72. A possible beneficial effect of HSP72 is discussed.

Acclimatization↗

Zinc acclimation and its effect on the zinc tolerance of Raphidocelis subcapitata and Chlorella vulgaris in laboratory experiments.

The effect of zinc acclimation of Raphidocelis subcapitata (syn. Selenastrum capricornutum) and Chlorella vulgaris on their sensitivity towards this metal was examined in a series of laboratory experiments. These two commonly used algal species were acclimated to 65 microg Zn/l and changes in zinc tolerance were monitored using standard growth inhibition tests. The chemically defined ISO medium was used as a control culture medium. Both species demonstrated a maximum increase in zinc tolerance of a factor of 3 after 100 days of acclimation. Shifts in the shape of the concentration-response curve due to acclimation were observed for R. subcapitata. Compared to non-acclimated algae, acclimated R. subcapitata exhibited higher growth rates in all zinc treatments as well as in the controls. This suggests that the use of ISO-medium results in sub-optimal growth due to zinc deficiency. These effects could not be demonstrated for C. vulgaris. The zinc tolerance of both species decreased significantly one week after returning the acclimated algae to control (ISO) medium. 72hEC50 values based on growth rate were two to four times higher than those calculated using biomass measurements. Algal toxicity test results, particularly if used for metal risk assessments, must not be conducted using nutrient deficient media.

Acclimatization↗

Differential scanning calorimetry and CD spectrometry of acclimation temperature-associated types of carp light meromyosin.

Differential scanning calorimetry and CD spectrometry were employed to study the thermal unfolding of light meromyosin (LMM) prepared from carp acclimated to different temperatures. The transition temperatures given by the major peaks at pH 8.0 in 0.6 M KCl for LMM from carp acclimated to 10 degrees C were 32.5 and 39.5 degrees C with the calorimetric enthalpies (DeltaHcal) of 269 and 52 kcal/mol, respectively. LMM from carp acclimated to 20 degrees C exhibited three peaks of transition temperatures at 34.5, 40.2, and 46.9 with DeltaHcal of 152, 20, and 10 kcal/mol, respectively. On the other hand, LMM from carp acclimated to 30 degrees C showed two different patterns. The first experiment gave two transition temperatures at 39.2 and 47.3 degrees C with DeltaHcal of 231 and 39 kcal/mol, respectively. The second series of experiments resulted in showing three peaks of 34.4, 39.5, and 47.5 degrees C with DeltaHcal of 117, 123, and 28 kcal/mol, respectively. N-terminal amino acid sequence analysis revealed that LMM at the second series of experiments with the 30 degrees C-acclimated carp contained component(s) predominant in the 20 degrees C-acclimated carp. Thermal unfolding responsible for these transition temperatures was well explained by melting of alpha-helices which could be determined by far-ultraviolet CD spectroscopy. These results clearly demonstrate that the 30 degrees C-acclimated carp contained the most thermostable LMM.

Acclimatization↗

Mass spectrometric approach for identifying putative plasma membrane proteins of Arabidopsis leaves associated with cold acclimation.

Although enhancement of freezing tolerance in plants during cold acclimation is closely associated with an increase in the cryostability of plasma membrane, the molecular mechanism for the increased cryostability of plasma membrane is still to be elucidated. In Arabidopsis, enhanced freezing tolerance was detectable after cold acclimation at 2 degrees C for as short as 1 day, and maximum freezing tolerance was attained after 1 week. To identify the plasma membrane proteins that change in quantity in response to cold acclimation, a highly purified plasma membrane fraction was isolated from leaves before and during cold acclimation, and the proteins in the fraction were separated with gel electrophoresis. We found that there were substantial changes in the protein profiles after as short as 1 day of cold acclimation. Subsequently, using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS), we identified 38 proteins that changed in quantity during cold acclimation. The proteins that changed in quantity during the first day of cold acclimation include those that are associated with membrane repair by membrane fusion, protection of the membrane against osmotic stress, enhancement of CO2 fixation, and proteolysis.

Acclimatization↗

Thermal acclimation of metabolic rate may be seasonally dependent in the subtropical anuran Latouche's frog (Rana latouchii, Boulenger).

We tested the hypothesis that the lack of metabolic thermal acclimation ability in tropical and subtropical amphibians is dependent on season and investigated the effects of body size, sex, time of day, and season on metabolic rates in Rana latouchii. The males were acclimated at 15 degrees, 20 degrees, and 25 degrees C, and their oxygen consumption was measured at 15 degrees, 20 degrees, 25 degrees, and 30 degrees C in all four seasons, with the exception that we did not measure oxygen consumption at 30 degrees C in winter frogs. We also acclimated the males at 30 degrees C in summer for investigating diel variation of metabolic rate. The females were acclimated at 20 degrees and 25 degrees C, and their oxygen consumption was measured at 15 degrees , 20 degrees , 25 degrees , and 30 degrees C in summer. Our results showed that metabolic rates of R. latouchii differed by time of day, season, and acclimation temperature but did not differ by sex if the results were adjusted for differences in body mass. Summer males exhibited a 26%-48% increase in metabolic rates from the lowest values in the seasons. There was a trend of increased oxygen consumption in cold-acclimated males, but it was significant only at 15 degrees and 25 degrees C in summer, autumn, and winter. These results support the hypothesis that thermal acclimation of metabolism is seasonally dependent, which has not been reported in other tropical and subtropical amphibians.

Acclimatization↗

Novel interrelationship between salicylic acid, abscisic acid, and PIP2-specific phospholipase C in heat acclimation-induced thermotolerance in pea leaves.

Increasing evidence suggests that heat acclimation and exogenous salicylic acid (SA) and abscisic acid (ABA) may lead to the enhancement of thermotolerance in plants. In this study, the roles that free SA, conjugated SA, ABA, and phosphatidylinositol-4,5-bisphosphate (PIP(2))-specific phospholipase C (PLC) play in thermotolerance development induced by heat acclimation (38 degrees C) were investigated. To evaluate their potential functions, three inhibitors of synthesis or activity were infiltrated into pea leaves prior to heat acclimation treatment. The results showed that the burst of free SA in response to heat acclimation could be attributed to the conversion of SA 2-O-D-glucose, the main conjugated form of SA, to free SA. Inhibition of ABA biosynthesis also resulted in a defect in the free SA peak during heat acclimation. In acquired thermotolerance assessment, the greatest weakness of antioxidant enzyme activity and the most severe heat injury (malondialdehyde content and degree of wilting) were found in pea leaves pre-treated with neomycin, a well-known inhibitor of PIP(2)-PLC activity. PsPLC gene expression was activated by exogenous ABA, SA treatments, and heat acclimation after pre-treatments with a SA biosynthesis inhibitor. From these results, PIP(2)-PLC appears to play a key role in free SA- and ABA-associated reinforcement of thermotolerance resulting from heat acclimation.

Abscisic Acid↗

Fast skeletal myosin isoforms in thermally acclimated carp.

Fast skeletal myosins were isolated from carp acclimated to 10 and 30 degrees C, and their structural and enzymatic properties were compared. Myosins in 0.5 M KCl were subjected to limited proteolysis by using various proteases including alpha-chymotrypsin, trypsin, and papain, and different SDS-PAGE patterns were seen for the 10- and 30 degrees C-acclimated myosins in all cases. Myosin subfragment-1 (S1) prepared from the 10 degrees C-acclimated myosin by alpha-chymotryptic digestion in 0.12 M NaCl showed higher acto-S1 Mg(2+)-ATPase activity and lower thermostability than S1 from the warm-acclimated myosin. The peptide maps and ATP-induced spectral changes of tryptophan fluorescence also showed an obvious difference between the two types of S1. Temperature acclimation further caused changes in the rod region of myosin, since the apparent sizes of light meromyosin were different from each other for the two types of myosin. Myosin from carp acclimated to 20 degrees C showed intermediate properties between those of the 10- and 30 degrees C-acclimated myosins. Myosin isoforms might be expressed in a temperature-dependent manner to compensate for the effect of seasonal environmental temperature variation on swimming ability.

Acclimatization↗

Changes in ABA and gene expression in cold-acclimated sugar maple.

To determine if cold acclimation of sugar maple (Acer saccharum Marsh.) is associated with specific changes in gene expression under natural hardening conditions, we compared bud and root translatable mRNAs of potted maple seedlings after cold acclimation under natural conditions and following spring dehardening. Cold-hardened roots and buds were sampled in January when tissues reached their maximum hardiness. Freezing tolerance, expressed as the lethal temperature for 50% of the tissues (LT(50)), was estimated at -17 degrees C for roots, and at lower than -36 degrees C for buds. Approximately ten transcripts were specifically synthesized in cold-acclimated buds, or were more abundant in cold-acclimated buds than in unhardened buds. Cold hardening was also associated with changes in translation. At least five translation products were more abundant in cold-acclimated buds and roots compared with unhardened tissues. Abscisic acid (ABA) concentration increased approximately tenfold in the xylem sap following winter acclimation, and the maximum concentration was reached just before maximal acclimation. We discuss the potential involvement of ABA in the observed modification of gene expression during cold hardening.

Journal Article↗

Carbon status constrains light acclimation in the cyanobacterium Synechococcus elongatus.

Acclimation to one environmental factor may constrain acclimation to another. Synechococcus elongatus (sp. PCC7942), growing under continuous light in high inorganic carbon (Ci; approximately 4 mm) and low-Ci (approximately 0.02 mm) media, achieve similar photosynthetic and growth rates under continuous low or high light. During acclimation from low to high light, however, high-Ci cells exploit the light increase by accelerating their growth rate, while low-Ci cells maintain the prelight shift growth rate for many hours, despite increased photosynthesis under the higher light. Under increased light, high-Ci cells reorganize their photosynthetic apparatus by shrinking the PSII pool and increasing Rubisco pool size, thus decreasing the photosynthetic source-to-sink ratio. Low-Ci cells also decrease their reductant source-to-sink ratio to a similar level as the high-Ci cells, but do so only by increasing their Rubisco pool. Low-Ci cells thus invest more photosynthetic reductant into maintaining their larger photosystem pool and increasing their Rubisco pool at the expense of population growth than do high-Ci cells. In nature, light varies widely over minutes to hours and is ultimately limited by daylength. Photosynthetic acclimation in S. elongatus occurs in both high and low Ci, but low-Ci cells require more time to achieve acclimation. Cells that can tolerate low Ci do so at the expense of slower photosynthetic acclimation. Such differences in rates of acclimation relative to rates of change in environmental parameters are important for predicting community productivity under variable environments.

Acclimatization↗

Nucleotide Levels Do Not Critically Determine Survival of Maize Root Tips Acclimated to a Low-Oxygen Environment.

We tested the hypothesis that ATP levels and energy charge determine the resistance of maize (Zea mays) root tips to anoxia. We focused on root tips of whole maize seedlings that had been acclimated to low O2 by exposure to an atmosphere of 3% (v/v) O2 in N2. Acclimated anoxic root tips characteristically have higher ATP levels and energy charge and survive longer under anoxia than nonacclimated tips. We poisoned intact, acclimated root tips with either fluoride or mannose, causing decreases in ATP and energy charge to values similar to or, in most cases, below those found in nonacclimated anoxic tips. With the exception of the highest fluoride concentration used, the poisoned, acclimated tips remained much more tolerant of anoxia than nonacclimated root tips. We conclude that high ATP and energy charge are not components critical for the survival of acclimated root tips during anoxia. The reduced nucleotide status in poisoned, acclimated root tips had little effect on cytoplasmic pH regulation during anoxia. This result indicates that in anoxic, acclimated root tips either cytoplasmic pH regulation is not dominated by ATP-dependent processes or these processes can continue in vivo largely independently of any changes in ATP levels in the physiological range. The role of glycolytic flux in survival under anoxia is discussed.

Journal Article↗