Tim Muir | Augustana College at Rock Island (original) (raw)

Papers by Tim Muir

International Journal of Molecular Sciences, 2011

International Journal of Molecular Sciences, 2011

Winter's advent invokes physiological adjustments that permit temperate ectotherms to cope with s... more Winter's advent invokes physiological adjustments that permit temperate ectotherms to cope with stresses such as food shortage, water deprivation, hypoxia, and hypothermia. We used liquid chromatography (LC) in combination with tandem mass spectrometry (MS/MS) quantitative isobaric (iTRAQ™) peptide mapping to assess variation in the abundance of hepatic proteins in summer-and winter-acclimatized wood frogs (Rana sylvatica), a northerly-distributed species that tolerates extreme dehydration and tissue freezing during hibernation. Thirty-three unique proteins exhibited strong seasonal lability. Livers of winter frogs had relatively high levels of proteins involved in cytoprotection, including heat-shock proteins and an antioxidant, and a reduced abundance of proteins involved in cell proliferation, protein synthesis, and mitochondrial function. They also exhibited altered levels of certain metabolic enzymes that participate in the biochemical reorganization associated with aphagia and reliance on energy reserves, as well as the freezing mobilization and post-thaw recovery of glucose, an important cryoprotective solute in freezing adaptation.

Physiological and Biochemical Zoology, 2010

Although many studies of ectothermic vertebrates have documented compensatory changes in cold har... more Although many studies of ectothermic vertebrates have documented compensatory changes in cold hardiness associated with changes of season, much less attention has been paid to adjustment of physiological functions and survival limits following more acute exposure to cold. We investigated the ability of hatchling painted turtles (Chrysemys picta) to increase cold hardiness in response to brief exposure to a subzero temperature. Winter-acclimated turtles were "cold conditioned" by chilling them in the supercooled (unfrozen) state to Ϫ7ЊC over a few days before returning them to 4ЊC. These turtles fared no better than control animals in resisting freezing when cooled in the presence or absence of ice and exogenous ice nuclei. Survival following tests of freeze tolerance (freezing for about 70 h; minimum body temperature, Ϫ3.75ЊC) was nominally higher in cold-conditioned turtles than in controls (36% vs. 13%, respectively), although the difference was not statistically significant. Of the survivors, cold-conditioned turtles apparently recovered sooner. Turtles subjected to cold shock (supercooling to Ϫ13ЊC for 24 h, followed by rewarming to 0ЊC) were strongly affected by cold conditioning: all controls died, but 50% of cold-conditioned turtles survived. We investigated potential mechanisms underlying the response to cold conditioning by measuring changes in levels of putative cryoprotectants. Plasma levels of glucose and lactate, but not urea, were higher in cold-conditioned turtles than in controls, although the combined increase in these solutes was only 23 mmol L Ϫ1 . Cold conditioning attenuated cold-shock injury to brain cells, as assessed using a vital-dye assay, suggesting a link between protection of the nervous system and cold hardiness at the organismal level.

Journal of Thermal Biology, 2013

The painted turtle (Chrysemys picta) is an especially useful organism in the study of metabolic r... more The painted turtle (Chrysemys picta) is an especially useful organism in the study of metabolic regulation during dormancy because it is sustained by finite energy reserves from hatching until emerging from its nest, about nine months later. In this study we subjected overwintering C. picta hatchlings to 4, 10, or 15 1C, temperatures simulating cold, mild, and warm winters, respectively, to investigate how various energy reserves are impacted by differential metabolic demands. An energy budget based on seasonal changes in caloric content showed that these turtles consumed an average of 0.39, 0.75, or 1.21 kJ g −1 , respectively, during the 6-month period of simulated hibernation. These estimates of energy use agreed reasonably well with estimates based solely on respirometric data. Unexpectedly, turtles in autumn contained little residual yolk, none of which was consumed by turtles in the cold-and mild-winter groups, this finding contradicting the widely held belief that residual yolk plays an important, direct role in the survival of turtles that overwinter inside their natal nest. By contrast, a marked reduction in dry mass of both liver and carcass attested to their importance in fueling metabolism and, indeed, catabolism of substrates from these components accounted for 31-52 and 35-63%, respectively, of the energetic cost of overwintering. The greater dependence on carcass reserves and relatively poor physiological condition of turtles in the mild-and warm-winter groups implies that metabolic demands imposed by high environmental temperatures would likely constrain post-emergence fitness.

Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2000

Many organisms endure extended periods of dormancy by depressing their metabolism, which effectiv... more Many organisms endure extended periods of dormancy by depressing their metabolism, which effectively prolongs the use of their endogenous energy stores. Though the mechanisms of hypometabolism are varied and incompletely understood, recent work suggests that urea accumulation in autumn and early winter contributes to reduced metabolism of hibernating wood frogs (Rana sylvatica). Urea accumulation during dormancy is a widespread phenomenon, and it has long been presumed that numerous species from diverse taxa benefit from its hypometabolic effect. To investigate the phylogenetic prevalence of urea-induced hypometabolism, we studied four species of urea accumulators from the clades Amphibia (Spea bombifrons and Ambystoma tigrinum), Reptilia (Malaclemys terrapin), and Gastropoda (Anguispira alternata), and one amphibian species (R. pipiens) that does not accumulate urea during dormancy. We measured rates of oxygen consumption ( _ V O 2 ) of excised organ samples from dormant animals in the presence or absence of physiological concentrations of urea. Three of the four urea-accumulating species had at least one organ whose _ V O 2 was significantly decreased by urea treatment. However, _ V O 2 of organs from R. pipiens, the one species tested that does not accumulate urea during dormancy, was not affected by urea treatment. Our results support the hypothesis that urea accumulation can reduce metabolic rate of dormant animals and provide a base for further investigation into the evolution of urea-induced hypometabolism.

Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2008

It has long been suspected that urea accumulation plays a key role in the induction or maintenanc... more It has long been suspected that urea accumulation plays a key role in the induction or maintenance of metabolic suppression during extended dormancy in animals from diverse taxa. However, little evidence supporting that hypothesis in living systems exists. We measured aerobic metabolism of isolated organs from the wood frog (Rana sylvatica) in the presence or absence of elevated urea at various temperatures using frogs acclimatized to different seasons. The depressive effect of urea on metabolism was not consistent across organs, seasons, or temperatures. None of the organs from summer frogs, which were tested at 201C, or from winter frogs tested at 41C were affected by urea treatment. However, liver, stomach, and heart from spring frogs tested at 41C had significantly lower metabolic rates when treated with urea as compared with control samples. Additionally, when organs from winter frogs were tested at 101C, metabolism was significantly decreased in urea-treated liver and stomach by 1515% and in urea-treated skeletal muscle by 1550%. Our results suggest that the presence of urea depresses the metabolism of living organs, and thereby reduces energy expenditure, but its effect varies with temperature and seasonal acclimatization. The impact of our findings may be wide ranging owing to the number of diverse organisms that accumulate urea during dormancy.

Journal of Comparative Physiology B, 2010

It has long been speculated that urea accumulated during seasonal dormancy contributes to metabol... more It has long been speculated that urea accumulated during seasonal dormancy contributes to metabolic depression. Recent work suggests urea can indeed act as a metabolic depressant during dormancy in a number of taxonomically diverse species of ectotherms. The mechanisms by which urea exerts its hypometabolic effect are unknown, but potentially stem from inhibition of mitochondrial respiration. We isolated mitochondria from Rana sylvatica skeletal muscle, an organ that is metabolically responsive to urea, and measured respiration rates in the absence or presence of 80 mmol l(-1) urea in the respiration medium. Because the effect of urea may be influenced by the intracellular milieu, in these experiments we varied substrate (pyruvate or palmitoylcarnitine), temperature (4, 10, or 15°C), and pH (6.8 or 7.4). Oxygen consumption of control and urea-treated mitochondria was sensitive to each of these variables, but neither state 3 nor state 4 respiration was reduced by urea treatment and, to the contrary, urea treatment slightly increased state 4 respiration at higher test temperatures. Although we did not test the efficacy of other incubation times or urea concentrations, the outcome of our experiment intimates that the urea-induced hypometabolism observed in hibernating R. sylvatica results from inhibition of energy-utilizing processes elsewhere in the cell, rather than a direct inhibition of mitochondrial respiration. Future investigation into urea's effects on non-mitochondrial metabolic pathways is necessary to uncover the mechanisms by which urea depresses metabolic rate.

Journal of Comparative Physiology B, 2007

Physiological responses to dehydration in amphibians are reasonably well documented, although lit... more Physiological responses to dehydration in amphibians are reasonably well documented, although little work has addressed this problem in hibernating animals. We investigated osmotic and metabolic responses to experimental manipulation of hydration state in the wood frog (Rana sylvatica), a terrestrial hibernator that encounters low environmental water potential during autumn and winter. In winter-conditioned frogs, plasma osmolality varied inversely with body water content (range 69-79%, fresh mass) primarily due to increases in sodium and chloride concentrations, as well as accumulation of glucose and urea. Decreased hydration was accompanied by a marked reduction in the resting rate of oxygen consumption, which was inversely correlated with plasma osmolality and urea concentration. In a separate experiment, resting rates of oxygen consumption in fully hydrated frogs receiving injections of saline or saline containing urea did not differ initially; however, upon dehydration, metabolic rates decreased sooner in the urea-loaded frogs than in control frogs. Our findings suggest an important role for urea, acting in concert with dehydration, in the metabolic regulation and energy conservation of hibernating R. sylvatica.

International Journal of Molecular Sciences, 2011

International Journal of Molecular Sciences, 2011

Winter's advent invokes physiological adjustments that permit temperate ectotherms to cope with s... more Winter's advent invokes physiological adjustments that permit temperate ectotherms to cope with stresses such as food shortage, water deprivation, hypoxia, and hypothermia. We used liquid chromatography (LC) in combination with tandem mass spectrometry (MS/MS) quantitative isobaric (iTRAQ™) peptide mapping to assess variation in the abundance of hepatic proteins in summer-and winter-acclimatized wood frogs (Rana sylvatica), a northerly-distributed species that tolerates extreme dehydration and tissue freezing during hibernation. Thirty-three unique proteins exhibited strong seasonal lability. Livers of winter frogs had relatively high levels of proteins involved in cytoprotection, including heat-shock proteins and an antioxidant, and a reduced abundance of proteins involved in cell proliferation, protein synthesis, and mitochondrial function. They also exhibited altered levels of certain metabolic enzymes that participate in the biochemical reorganization associated with aphagia and reliance on energy reserves, as well as the freezing mobilization and post-thaw recovery of glucose, an important cryoprotective solute in freezing adaptation.

Physiological and Biochemical Zoology, 2010

Although many studies of ectothermic vertebrates have documented compensatory changes in cold har... more Although many studies of ectothermic vertebrates have documented compensatory changes in cold hardiness associated with changes of season, much less attention has been paid to adjustment of physiological functions and survival limits following more acute exposure to cold. We investigated the ability of hatchling painted turtles (Chrysemys picta) to increase cold hardiness in response to brief exposure to a subzero temperature. Winter-acclimated turtles were "cold conditioned" by chilling them in the supercooled (unfrozen) state to Ϫ7ЊC over a few days before returning them to 4ЊC. These turtles fared no better than control animals in resisting freezing when cooled in the presence or absence of ice and exogenous ice nuclei. Survival following tests of freeze tolerance (freezing for about 70 h; minimum body temperature, Ϫ3.75ЊC) was nominally higher in cold-conditioned turtles than in controls (36% vs. 13%, respectively), although the difference was not statistically significant. Of the survivors, cold-conditioned turtles apparently recovered sooner. Turtles subjected to cold shock (supercooling to Ϫ13ЊC for 24 h, followed by rewarming to 0ЊC) were strongly affected by cold conditioning: all controls died, but 50% of cold-conditioned turtles survived. We investigated potential mechanisms underlying the response to cold conditioning by measuring changes in levels of putative cryoprotectants. Plasma levels of glucose and lactate, but not urea, were higher in cold-conditioned turtles than in controls, although the combined increase in these solutes was only 23 mmol L Ϫ1 . Cold conditioning attenuated cold-shock injury to brain cells, as assessed using a vital-dye assay, suggesting a link between protection of the nervous system and cold hardiness at the organismal level.

Journal of Thermal Biology, 2013

The painted turtle (Chrysemys picta) is an especially useful organism in the study of metabolic r... more The painted turtle (Chrysemys picta) is an especially useful organism in the study of metabolic regulation during dormancy because it is sustained by finite energy reserves from hatching until emerging from its nest, about nine months later. In this study we subjected overwintering C. picta hatchlings to 4, 10, or 15 1C, temperatures simulating cold, mild, and warm winters, respectively, to investigate how various energy reserves are impacted by differential metabolic demands. An energy budget based on seasonal changes in caloric content showed that these turtles consumed an average of 0.39, 0.75, or 1.21 kJ g −1 , respectively, during the 6-month period of simulated hibernation. These estimates of energy use agreed reasonably well with estimates based solely on respirometric data. Unexpectedly, turtles in autumn contained little residual yolk, none of which was consumed by turtles in the cold-and mild-winter groups, this finding contradicting the widely held belief that residual yolk plays an important, direct role in the survival of turtles that overwinter inside their natal nest. By contrast, a marked reduction in dry mass of both liver and carcass attested to their importance in fueling metabolism and, indeed, catabolism of substrates from these components accounted for 31-52 and 35-63%, respectively, of the energetic cost of overwintering. The greater dependence on carcass reserves and relatively poor physiological condition of turtles in the mild-and warm-winter groups implies that metabolic demands imposed by high environmental temperatures would likely constrain post-emergence fitness.

Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2000

Many organisms endure extended periods of dormancy by depressing their metabolism, which effectiv... more Many organisms endure extended periods of dormancy by depressing their metabolism, which effectively prolongs the use of their endogenous energy stores. Though the mechanisms of hypometabolism are varied and incompletely understood, recent work suggests that urea accumulation in autumn and early winter contributes to reduced metabolism of hibernating wood frogs (Rana sylvatica). Urea accumulation during dormancy is a widespread phenomenon, and it has long been presumed that numerous species from diverse taxa benefit from its hypometabolic effect. To investigate the phylogenetic prevalence of urea-induced hypometabolism, we studied four species of urea accumulators from the clades Amphibia (Spea bombifrons and Ambystoma tigrinum), Reptilia (Malaclemys terrapin), and Gastropoda (Anguispira alternata), and one amphibian species (R. pipiens) that does not accumulate urea during dormancy. We measured rates of oxygen consumption ( _ V O 2 ) of excised organ samples from dormant animals in the presence or absence of physiological concentrations of urea. Three of the four urea-accumulating species had at least one organ whose _ V O 2 was significantly decreased by urea treatment. However, _ V O 2 of organs from R. pipiens, the one species tested that does not accumulate urea during dormancy, was not affected by urea treatment. Our results support the hypothesis that urea accumulation can reduce metabolic rate of dormant animals and provide a base for further investigation into the evolution of urea-induced hypometabolism.

Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2008

It has long been suspected that urea accumulation plays a key role in the induction or maintenanc... more It has long been suspected that urea accumulation plays a key role in the induction or maintenance of metabolic suppression during extended dormancy in animals from diverse taxa. However, little evidence supporting that hypothesis in living systems exists. We measured aerobic metabolism of isolated organs from the wood frog (Rana sylvatica) in the presence or absence of elevated urea at various temperatures using frogs acclimatized to different seasons. The depressive effect of urea on metabolism was not consistent across organs, seasons, or temperatures. None of the organs from summer frogs, which were tested at 201C, or from winter frogs tested at 41C were affected by urea treatment. However, liver, stomach, and heart from spring frogs tested at 41C had significantly lower metabolic rates when treated with urea as compared with control samples. Additionally, when organs from winter frogs were tested at 101C, metabolism was significantly decreased in urea-treated liver and stomach by 1515% and in urea-treated skeletal muscle by 1550%. Our results suggest that the presence of urea depresses the metabolism of living organs, and thereby reduces energy expenditure, but its effect varies with temperature and seasonal acclimatization. The impact of our findings may be wide ranging owing to the number of diverse organisms that accumulate urea during dormancy.

Journal of Comparative Physiology B, 2010

It has long been speculated that urea accumulated during seasonal dormancy contributes to metabol... more It has long been speculated that urea accumulated during seasonal dormancy contributes to metabolic depression. Recent work suggests urea can indeed act as a metabolic depressant during dormancy in a number of taxonomically diverse species of ectotherms. The mechanisms by which urea exerts its hypometabolic effect are unknown, but potentially stem from inhibition of mitochondrial respiration. We isolated mitochondria from Rana sylvatica skeletal muscle, an organ that is metabolically responsive to urea, and measured respiration rates in the absence or presence of 80 mmol l(-1) urea in the respiration medium. Because the effect of urea may be influenced by the intracellular milieu, in these experiments we varied substrate (pyruvate or palmitoylcarnitine), temperature (4, 10, or 15°C), and pH (6.8 or 7.4). Oxygen consumption of control and urea-treated mitochondria was sensitive to each of these variables, but neither state 3 nor state 4 respiration was reduced by urea treatment and, to the contrary, urea treatment slightly increased state 4 respiration at higher test temperatures. Although we did not test the efficacy of other incubation times or urea concentrations, the outcome of our experiment intimates that the urea-induced hypometabolism observed in hibernating R. sylvatica results from inhibition of energy-utilizing processes elsewhere in the cell, rather than a direct inhibition of mitochondrial respiration. Future investigation into urea's effects on non-mitochondrial metabolic pathways is necessary to uncover the mechanisms by which urea depresses metabolic rate.

Journal of Comparative Physiology B, 2007

Physiological responses to dehydration in amphibians are reasonably well documented, although lit... more Physiological responses to dehydration in amphibians are reasonably well documented, although little work has addressed this problem in hibernating animals. We investigated osmotic and metabolic responses to experimental manipulation of hydration state in the wood frog (Rana sylvatica), a terrestrial hibernator that encounters low environmental water potential during autumn and winter. In winter-conditioned frogs, plasma osmolality varied inversely with body water content (range 69-79%, fresh mass) primarily due to increases in sodium and chloride concentrations, as well as accumulation of glucose and urea. Decreased hydration was accompanied by a marked reduction in the resting rate of oxygen consumption, which was inversely correlated with plasma osmolality and urea concentration. In a separate experiment, resting rates of oxygen consumption in fully hydrated frogs receiving injections of saline or saline containing urea did not differ initially; however, upon dehydration, metabolic rates decreased sooner in the urea-loaded frogs than in control frogs. Our findings suggest an important role for urea, acting in concert with dehydration, in the metabolic regulation and energy conservation of hibernating R. sylvatica.