Ingestion of juniper foliage reduces metabolic rates in woodrat (Neotoma) herbivores

Ingestion of plant secondary compounds by herbivores is predicted to increase resting or basal metabolic rates. We tested this hypothesis with two species of woodrat herbivores, Neotoma stephensi and Neotoma albigula, consuming diets of juniper (Juniperus monosperma), which is rich in plant secondary compounds. In nature, N. stephensi specializes on juniper, whereas N. albigula consumes a variety of plant species including juniper. We measured resting metabolic rates (RMR) of woodrats on control, 25% juniper and a treatment containing the maximum tolerable dose of juniper (50% juniper for N. albigula and 70% juniper for N. stephensi). Ingestion of a juniper diet resulted in decreased RMR in both species of woodrats. We propose several potential mechanisms for metabolic depression of Neotoma on juniper diets. Our novel results underscore the need for more studies utilizing plant-based diets to determine the general effect of plant secondary compounds on metabolic rates of herbivores.     

Ambient temperature influences diet selection and physiology of an herbivorous mammal, Neotoma albigula

The whitethroat woodrat (Neotoma albigula) eats juniper (Juniperus monosperma), but the amount of juniper in its diet varies seasonally. We tested whether changes in juniper consumption are due to changes in ambient temperature and what the physiological consequences of consuming plant secondary compounds (PSCs) at different ambient temperatures might be. Woodrats were acclimated to either 20 degrees C or 28 degrees C. Later, they were given two diets to choose from (50% juniper and a nontoxic control) for 7 d. Food intake, resting metabolic rate (RMR), and body temperature (T(b)) were measured over the last 2 d. Woodrats at 28 degrees C ate significantly less juniper, both proportionally and absolutely, than woodrats at 20 degrees C. RMRs were higher for woodrats consuming juniper regardless of ambient temperature, and T(b) was higher for woodrats consuming juniper at 28 degrees C than for woodrats eating control diet at 28 degrees C. Thus, juniper consumption by N. albigula is influenced by ambient temperature. We conclude that juniper may influence thermoregulation in N. albigula in ways that are helpful at low temperatures but harmful at warmer temperatures in that juniper PSCs may be more toxic at warmer temperatures. The results suggest that increases in ambient temperature associated with climate change could significantly influence foraging behavior of mammalian herbivores.     

Intestinal capacity of P-glycoprotein is higher in the juniper specialist, Neotoma stephensi, than the sympatric generalist, Neotoma albigula

Permeability-glycoprotein (Pgp) is a membrane-bound, ATP-dependent, transport protein that excludes many cytotoxic compounds including plant metabolites and pollutants from the barrier epithelia of many tissues including the small intestine. We hypothesized that intestinal Pgp capacity would be higher in Neotoma stephensi, a specialist on Juniperus monosperma known to be high in plant toxins, than the sympatric generalist, Neotoma albigula, which consumes juniper in the field, but is unable to tolerate a high juniper diet. We measured Pgp activity as the difference in accumulation of a known Pgp substrate, digoxin, between everted sections of small intestine exposed to ethanol vehicle control and a maximal level of a known competitive inhibitor of Pgp, cyclosporin A. We estimated intestinal capacity by averaging Pgp activity along the intestine and multiplying by total small intestine mass. These first measures of Pgp in wild mammals show a significant difference among species with the juniper specialist, N. stephensi, exhibiting a 2.4 fold higher capacity than the generalist, N. albigula. This result suggests that Pgp may play a role in the ability of N. stephensi to tolerate juniper.     

Xenobiotic metabolism of plant secondary compounds in juniper (Juniperus monosperma) by specialist and generalist woodrat herbivores, genus Neotoma

Mammalian herbivores routinely consume diets laden with often-toxic xenobiotics, yet the manner in which mammalian herbivores detoxify these plant secondary compounds (PSC) is largely unknown. Theory predicts that specialists rely more heavily on functionalization pathways whereas generalists rely on conjugation pathways to metabolize PSC in their diet. We took a pharmacological approach to determine how a specialist (Neotoma stephensi) of juniper foliage (Juniperus monosperma) and a generalist (N. albigula) may process the same dietary PSC. We investigated the xenobiotic metabolizing enzymes of the specialist and generalist on a control diet and a low (25%) juniper diet. We also examined enzyme activities in the specialist on a high (70%) juniper diet. We assayed for cytochrome P450 concentration and biotransformation activities of three specific cytochrome P450 isozymes (CYP1A, CYP2B, CYP3A), NAD(P)H:quinone oxidoreductase, glutathione conjugation, sulfation and glucuronidation. Results provide partial evidence for the hypothesis in that the specialist and generalist consuming juniper at a level similar to their natural diet, differ in the level of conjugation enzyme activity with generalists having higher activity overall than specialists.     

A specialist herbivore (Neotoma stephensi) absorbs fewer plant toxins than does a generalist (Neotoma albigula)

Detoxification capacity of enzymes in the liver is thought to be the primary factor governing dietary toxin intake by mammalian herbivores. Recently, toxin absorption in the gut was proposed as an alternative process that also influences toxin intake. We examined the role of the gut in regulating toxin absorption by quantifying excretion of a plant secondary compound in the feces. We hypothesized that specialists have a greater capacity to reduce intestinal absorption of toxins than do generalists. To test this hypothesis, we compared fecal excretion of alpha-pinene in specialist (Neotoma stephensi) and generalist (Neotoma albigula) woodrats. Alpha-pinene is the most abundant monoterpene in Juniperus monosperma, which occurs in the natural diet of both woodrat species. Woodrats were fed alpha-pinene in diets containing juniper foliage for 3 wk and, in a separate experiment, were given a single oral dose of alpha-pinene. Feces were collected from animals at the end of each experiment and analyzed for alpha-pinene concentration using gas chromatography. Both woodrat species excreted unchanged alpha-pinene in the feces. However, specialist woodrats excreted 40% more alpha-pinene per unit ingested from a juniper diet and excreted nearly four times a greater percentage of an oral dose of alpha-pinene compared with generalists.     

Molecular systematics of the genus Neotoma

DNA sequences from the mitochondrial DNA cytochrome-b gene were used to infer the systematic relationships of 13 species of wood rats (genus Neotoma). Parsimony, likelihood, and neighbor-joining analyses produced similar topologies in most cases and produced six systematic conclusions. First, results of previous studies were supported in the recognition of N. floridana magister as distinct species (N. magister). Second, evidence was provided for the recognition of cryptic species within N. albigula (N. albigula and N. leucodon) and N. mexicana (N. mexicana, N. isthmica, and N. picta). Third, the subgenus Neotoma is composed of four species groups (floridana, lepida, mexicana, and micropus). Fourth, support was provided for placement of N. stephensi within the lepida species group. Fifth, support was provided for the recognition of Hodomys as a separate genus, sister to Xenomys. Sixth, support for the elevation of the subgenus Teonoma to generic status is discussed.     

Liver biotransforming enzymes in woodrats Neotoma stephensi (Muridae)

Mammalian herbivores are exposed to extremely high levels of plant secondary compounds naturally present in their diet. It has been speculated that specialist herbivores should express a unique pattern of biotransforming enzymes to permit the consumption of a single species of toxic plant. Specifically, specialists should rely on pathways that effectively biotransform the toxins they routinely encounter in their diet. We examined the hepatic mRNA expression and activity or content of biotransforming enzymes in the specialist herbivorous woodrat, Neotoma stephensi, and compared results to those of laboratory rats (Sprague-Dawley strain Rattus norvegicus). In addition, we investigated the role of alpha-pinene, a specific plant toxin present in the diet of N. stephensi on the mRNA expression pattern and activity or content of biotransforming enzymes in Sprague-Dawley rats. Overall, the levels of functionalization enzyme activity and mRNA were found to be higher in specialists, while glucuronidation enzyme activity and mRNA were lower. These results support predictions that specialist herbivores rely more on functionalization biotransformation pathways rather than glucuronidation pathways.     

Role of hyperthyroidism in increased thermogenesis in the cold-acclimated Syrian hamster

The oxygen consumption of cold-acclimated Syrian hamsters (measured in pentobarbital-anaesthetized animals) was 59% greater than that of warm-acclimated hamsters. Upon return of the cold-acclimated hamsters to 24 degrees C, the elevated metabolic rate declined slowly, with half-life of approximately 1 day. The increase correlated well with our previous finding of a four-fold increase in serum 3,5,3'-triiodothyronine concentration in cold-acclimated hamsters and the slow decline in concentration of this hormone during deacclimation. Daily administration of T3 (for 7 days) to warm-acclimated hamsters at 24 degrees C resulted in a similar increase in oxygen consumption that persisted 1 day after the last injection and had disappeared by 7 days after the last injection. We conclude that the high concentration of serum triiodothyronine in the cold-acclimated hamster exerts a thermogenic effect that probably contributes to thermoregulatory thermogenesis in the cold.     

Strategies in herbivory by mammals revisited: The role of liver metabolism in a juniper specialist (Neotoma stephensi) and a generalist (Neotoma albigula)

Although herbivory is widespread among mammals, few species have adopted a strategy of dietary specialization. Feeding on a single plant species often exposes herbivores to high doses of plant secondary metabolites (PSMs), which may exceed the animal's detoxification capacities. Theory predicts that specialists will have unique detoxification mechanisms to process high levels of dietary toxins. To evaluate this hypothesis, we compared liver microsomal metabolism of a juniper specialist, Neotoma stephensi (diet >85% juniper), to a generalist, N. albigula (diet ≤30% juniper). Specifically, we quantified the concentration of a key detoxification enzyme, cytochrome P450 2B (CYP2B) in liver microsomes, and the metabolism of α‐pinene, the most abundant terpene in the juniper species consumed by the specialist woodrat. In both species, a 30% juniper diet increased the total CYP2B concentration (2–3×) in microsomes and microsomal α‐pinene metabolism rates (4‐fold). In N. stephensi, higher levels of dietary juniper (60% and 100%) further induced CYP2B and increased metabolism rates of α‐pinene. Although no species‐specific differences in metabolism rates were observed at 30% dietary juniper, total microsomal CYP2B concentration was 1.7× higher in N. stephensi than in N. albigula (p < .01), suggesting N. stephensi produces one or more variant of CYP2B that is less efficient at processing α‐pinene. In N. stephensi, the rates of α‐pinene metabolism increased with dietary juniper and were positively correlated with CYP2B concentration. The ability of N. stephensi to elevate CYP2B concentration and rate of α‐pinene metabolism with increasing levels of juniper in the diet may facilitate juniper specialization in this species.     

Differential expression and activity of catechol-O-methyl transferase (COMT) in a generalist (Neotoma albigula) and juniper specialist (Neotoma stephensi) woodrat

Mammalian herbivores, particularly dietary specialists must have an efficient means to metabolize the high doses of plant secondary compounds they consume. We found previously that Neotoma stephensi, a juniper specialist, upregulated catechol-O-methyl transferase (COMT) mRNA almost seven fold in response to an ecologically relevant diet (70% juniper). To further investigate the relevance of this enzyme with respect to juniper metabolism, we compared the protein expression, activity and kinetics of the two forms of COMT, soluble (S-COMT) and membrane bound (MB-COMT), in the blood, kidneys and liver of N. stephensi on its natural juniper diet to that of N. stephensi fed an experimental diet of 70% juniper as well as a non-toxic control diet under laboratory conditions. In addition, we compared these results to that of Neotoma albigula, a generalist species, which consumes a diet of 25% juniper in the wild. The specialist consuming juniper under both field and laboratory conditions had increased S-COMT expression and activity in their livers and kidneys, and increased S-COMT activity in their blood compared to the specialist and generalist fed the control diet. The specialist showed expression and activity of S-COMT in their kidneys that was as high as or higher than that in their livers. The generalist had an elevated V_max for MB-COMT compared to the specialist that resulted in higher activity for MB-COMT than the specialist despite lower expression of MB-COMT in the generalist's livers and kidneys. This high activity MB-COMT may be in part responsible for differences in the behaviors of the generalist compared to the specialist. We conclude that S-COMT is important in the specialist's ability to consume high levels of juniper.     

Fatty acid and sterol synthesis by hepatocytes of thermally acclimated rainbow trout (Salmo gairdneri).

Incorporation of tritium from tritiated water into lipid fractions was measured in isolated hepatocytes from rainbow trout (Salmo gairdneri) acclimated to 5 degrees C and 20 degrees C. Hepatocytes from cold-acclimated trout exhibited significantly higher rates of tritium incorporation into both fatty acid and sterol fractions at assay temperatures of 15 degrees C and 20 degrees C than did hepatocytes from warm-acclimated trout. Tritium incorporation into the fatty acid fraction was nearly temperature independent in hepatocytes from warm-acclimated trout (Q10 = 1.39) but markedly temperature dependent (Q10 = 2.63) in hepatocytes from cold-acclimated trout; in contrast, rates of sterol synthesis were more temperature dependent in warm-acclimated trout. At 5 degrees C, fatty acid lipogenesis comprised a significantly greater percentage of the total tritium incorporation in hepatocytes from warm-acclimated trout and the percentage of total lipogenesis attributable to fatty acids decreased significantly in warm-acclimated trout as the assay temperature increased; the opposite trends were observed in cold-acclimated trout.     

Facultative hypothermic responses in an Afrotropical arid-zone passerine,the red-headed finch (<Emphasis Type="Italic">Amadina erythrocephala</Emphasis>)

We investigated thermoregulation and facultative hypothermic responses to food deprivation in the red-headed finch (Amadina erythrocephala), a 22-g passerine endemic to the arid regions of southern Africa. We predicted that, like most other passerines investigated, A. erythrocephala exhibits shallow rest-phase hypothermia, but not torpor. We observed significant reductions in rest-phase energy expenditure and body temperature (Tb) in response to restricted feeding. The maximum extent of Tb reduction (ca. 5 degrees C) and energy savings (ca. 10%) were consistent with those reported for a number of other passerine species. The lowest Tb we observed in a bird able to arouse spontaneously was 34.8 degrees C. The parameters of facultative hypothermic responses in A. erythrocephala were indicative of shallow rest-phase hypothermia, but not torpor. The limited available data on hypothermic responses in passerines suggest that many species do not possess the capacity for torpor. In passerines, torpor appears to be restricted to a few nectarivores and aerial insectarivores, and may have evolved independently of the torpor observed in non-passerine taxa such as the Trochiliformes and Caprimulgidae. The basal metabolic rate (BMR) of A. erythrocephala was 30-46% lower than predicted by various allometric equations, but was similar to the predicted BMR for a 22-g desert bird.     

Cold acclimation increases basal heart rate but decreases its thermal tolerance in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss, Walbaum) were acclimated to 4 degrees C and 17 degrees C for more than 4 weeks and heart rate was determined in the absence and presence of adrenaline to see how thermal adaptation influences basal heart rate and its beta-adrenergic control in a eurythermal fish species. The basal heart rate in vitro was higher in cold-acclimated than warm-acclimated rainbow trout at temperatures below 17 degrees C. On the other hand, adaptation to cold decreased thermal tolerance of heart rate so that the maximal heart rates were achieved at 17 degrees C (75 +/- 4 bpm) and 24 degrees C (88 +/- 2 bpm) in cold-acclimated and warm-acclimated trout, respectively. Beta-adrenergic response of the heart was enhanced by cold-adaptation, since adrenaline (100 nmol l(-1)) caused stronger stimulation of heart rate in cold-acclimated (29 +/- 14%) than in warm-acclimated fish (10 +/- 1%; P = 0.03). Furthermore, adrenaline strongly opposed the temperature-dependent deterioration of force production in cold-acclimated trout but not in warm-acclimated trout. The results indicate that adaptation to cold increases basal heart rate but decreases its thermal tolerance in rainbow trout. Cold acclimation up-regulates the beta-adrenergic system, and beta-adrenoceptor activation seems to provide cardioprotection against high temperatures in the cold-adapted rainbow trout.     

Ambient temperature does not affect fuelling rate in absence of digestive constraints in long-distance migrant shorebird fuelling up in captivity

Pre-flight fuelling rates in free-living red knots Calidris canutus, a specialized long-distance migrating shorebird species, are positively correlated with latitude and negatively with temperature. The single published hypothesis to explain these relationships is the heat load hypothesis that states that in warm climates red knots may overheat during fuelling. To limit endogenous heat production (measurable as basal metabolic rate BMR), birds would minimize the growth of digestive organs at a time they need. This hypothesis makes the implicit assumption that BMR is mainly driven by digestive organ size variation during pre-flight fuelling. To test the validity of this assumption, we fed captive knots with trout pellet food, a diet previously shown to quickly lead to atrophied digestive organs, during a fuelling episode. Birds were exposed to two thermal treatments (6 and 24°C) previously shown to generate different fuelling rates in knots. We made two predictions. First, easily digested trout pellet food rather than hard-shelled prey removes the heat contribution of the gut and would therefore eliminate an ambient temperature effect on fuelling rate. Second, if digestive organs were the main contributors to variations in BMR but did not change in size during fuelling, we would expect no or little change in BMR in birds fed ad libitum with trout pellets. We show that cold-acclimated birds maintained higher body mass and food intake (8 and 51%) than warm-acclimated birds. Air temperature had no effect on fuelling rate, timing of fuelling, timing of peak body mass or BMR. During fuelling, average body mass increased by 32% while average BMR increased by 15% at peak of mass and 26% by the end of the experiment. Our results show that the small digestive organs characteristic of a trout pellet diet did not prevent BMR from increasing during premigratory fuelling. Our results are not consistent with the heat load hypothesis as currently formulated.     

New host and locality records of coccidia (Apicomplexa: Eimeriidae) from rodents in the southwestern and western United States.

One hundred forty-seven murid and heteromyid rodents were collected from various sites in the southwestern and western United States (Arizona, Colorado, New Mexico, Texas, and Utah) and Baja California Norte, Mexico, and their feces were examined for coccidial parasites. Of these, 53 (36%) were infected with at least 1 coccidian; 45 of 53 (85%) of the infected rodents harbored only 1 species of coccidian. Infected rodents included: 10 of 22 (45%) Neotoma albigula, 3 of 11 (27%) Neotoma floridana, 2 of 14 (14%) Neotoma lepida, 15 of 29 (52%) Neotoma micropus, 5 of 8 (63%) Peromyscus crinitis, 6 of 6 (100%) Peromyscus difficilis, 1 of 2 (50%) Peromyscus eremicus, 9 of 34 (26%) Sigmodon hispidis, and 2 of 3 (67%) Sigmodon ochrognathus; 4 Neotoma cinerea, 3 Neotoma devia, 3 Neotoma mexicana, 1 Peromyscus maniculatus, 1 Onychomys leucogaster, 1 Onychomys torridus, 3 Chaetodipus fallax, and 2 Chaetodipus penicillatus were negative. Although no new species was found, the following coccidians were identified from infected rodents: Eimeria albigulae from N. albigula, N. floridana, and N. micropus, Eimeria antonellii from N. albigula and N. micropus, Eimeria ladronensis from N. albigula, N. floridana, N. lepida, and N. micropus, Eimeria arizonensis and Eimeria lachrymalis from P. crinitis and P. difficilis, Eimeria lachrymalis from P. eremicus, Eimeria tuskeegensis from S. ochrognathus, and Eimeria roperi, Eimeria sigmodontis, Eimeria tuskeegensis, Eimeria webbae, and an unidentified species of Eimeria from S. hispidis. This report documents 12 new host and several distributional records for Eimeria species from murid rodents in Arizona, Texas, and Utah.     

Temperature acclimation modifies sinoatrial pacemaker mechanism of the rainbow trout heart

The hypothesis of pacemaker level origin of thermal compensation in heart rate was tested by recording action potentials (AP) in intact sinoatrial tissue and enzymatically isolated pacemaker cells of rainbow trout acclimated at 4 degrees C (cold) and 18 degrees C (warm). With electrophysiological recordings, the primary pacemaker was located at the base of the sinoatrial valve, where a morphologically distinct ring of tissue comprising myocytes and neural elements was found by histological examination. Intrinsic beating rate of this pacemaker was higher in cold-acclimated (46 +/- 6 APs/min) than warm-acclimated trout (38 +/- 3 APs/min; P < 0.05), and a similar difference was seen in beating rate of isolated pacemaker cells (44 +/- 6 vs. 38 +/- 6 APs/min; P < 0.05), supporting the hypothesis that thermal acclimation modifies the intrinsic pacemaker mechanism of fish heart. Inhibition of sarcoplasmic reticulum (SR) with 10 microM ryanodine and 1 microM thapsigargin did not affect heart rate in either warm- or cold-acclimated trout at 11 degrees C but reduced heart rate in warm-acclimated trout from 74 +/- 2 to 42 +/- 6 APs/min (P < 0.05) at 18 degrees C. At 11 degrees C, a half-maximal blockade of the delayed rectifier K+ current (I(Kr)) with 0.1 microM E-4031 reduced heart rate more in warm-acclimated (from 45 +/- 1 to 24 +/- 5 APs/min) than cold-acclimated trout (56 +/- 3 vs. 48 +/- 2 APs/min), whereas I(Kr) density was higher and AP duration less in cold-acclimated trout (P > 0.05). Collectively, these findings suggest that a cold-induced increase in AP discharge frequency is at least partly due to higher density of the I(Kr) in the cold-acclimated trout, whereas contribution of SR Ca2+ release to thermal compensation of heart rate is negligible.     

Cross-transmission studies with Eimeria arizonensis-like oocysts (Apicomplexa) in New World rodents of the Genera baiomys, Neotoma, Onychomys, Peromyscus, and Reithrodontomys (Muridae).

Cross-transmission experiments were performed using oocysts of an Eimeria arizonensis-like coccidian from Peromyscus leucopus and Peromyscus truei, an E. arizonensis-like coccidian from Reithrodontomys fulvescens, Eimeria baiomysis and Eimeria taylori from Baiomys taylori, Eimeria albigulae from Neotoma albigula, and Eimeria onychomysis from Onychomys spp., between representatives of the above host genera. The E. arizonensis-like coccidian from R. fulvescens infected Reithrodontomys megalotis, Reithrodontomys montanus, and Peromyscus leucopus. Oocysts of E. arizonensis from P. leucopus could be transmitted to both P. leucopus and R. megalotus. Oocysts of E. baiomysis and E. taylori infected only B. taylori. Oocysts of E. arizonensis from P. truei infected P. truei but not Neotoma mexicana or Onychomys leucogaster. Oocysts of E. albigulae from N. albigula were infective for N. mexicana but not for P. truei or O. leucogaster. Oocysts of E. onychomysis from Onychomys spp. infected O. leucogaster but not N. mexicana or P. truei. These results demonstrate that Peromyscus and Reithrodontomys, genera known to be related very closely evolutionarily, are capable of sharing E. arizonensis, whereas morphologically similar coccidians (E. albigulae, E. baiomysis, and E. onychomysis) from more distantly related hosts, are probably distinct and more stenoxenous. This also is the first report of coccidians infecting species of Reithrodontomys.     

Basal metabolic rate, food intake, and body mass in cold- and warm-acclimated Garden Warblers

We address the question of whether physiological flexibility in relation to climate is a general feature of the metabolic properties of birds. We tested this hypothesis in hand-raised Garden Warblers (Sylvia borin), long-distance migrants, which normally do not experience great temperature differences between summer and winter. We maintained two groups of birds under cold and warm conditions for 5 months, during which their body mass and food intake were monitored. When relatedness (siblings vs. non-siblings) of the experimental birds was taken into account, body mass in cold-acclimated birds was higher than in warm-acclimated birds. BMR, measured at the end of the 5-month temperature treatment, was also higher in the cold- than the warm-acclimated group. Migrant birds thus seem to be capable of the same metabolic cold-acclimation response as has been reported in resident birds. The data support the hypothesis that physiological flexibility is a basic trait of the metabolic properties of birds.     

Behavioral thermoregulation by hypoxic rats.

To address whether a shift in hypothalamic thermal setpoint might be a significant factor in induction of hypoxic hypothermia, behavioral thermoregulation was examined in 7 female Sprague-Dawley rats implanted with radiotelethermometers for deep body temperature (Tb) measurement in a thermocline during normoxia (PO2 = 125 torr) and hypoxia (PO2 = 60 torr). Normoxic rats (TNox) selected a mean ambient temperature of 19.7 +/- 1.4 (SE) degrees C and maintained Tb at 37.0 +/- 0.2 degrees C. Hypoxic rats selected a significantly higher ambient temperature (THox = 28.6 +/- 2.2 degrees C) but maintained Tb significantly lower at 35.5 +/- 0.3 degrees C. Without a thermal gradient (ambient temperature = 25 degrees C), Tb during hypoxia was 35.4 +/- 0.4 degrees C. The maintenance of a lower body temperature during hypoxia through behavioral thermoregulation despite having warmer temperatures available supports the hypothesis that the thermoregulatory setpoint of hypoxic rats is shifted to promote thermoregulation at a lower Tb, effectively reducing oxygen demand when oxygen supply is limited.     

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.