scholarly journals Mitochondria and the Frozen Frog

Antioxidants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 543
Author(s):  
Janet M. Storey ◽  
Shaobo Wu ◽  
Kenneth B. Storey
Keyword(s):  
Cell Volume ◽  
Ischemia Reperfusion ◽  
Enzyme Regulation ◽  
Rana Sylvatica ◽  
Volume Reduction ◽  
Whole Body ◽  
Amphibian Species ◽  
Wood Frogs ◽  
16S Rna ◽  
Total Body

The wood frog, Rana sylvatica, is the best-studied of a small group of amphibian species that survive whole body freezing during the winter months. These frogs endure the freezing of 65–70% of their total body water in extracellular ice masses. They have implemented multiple adaptations that manage ice formation, deal with freeze-induced ischemia/reperfusion stress, limit cell volume reduction with the production of small molecule cryoprotectants (glucose, urea) and adjust a wide variety of metabolic pathways for prolonged life in a frozen state. All organs, tissues, cells and intracellular organelles are affected by freeze/thaw and its consequences. This article explores mitochondria in the frozen frog with a focus on both the consequences of freezing (e.g., anoxia/ischemia, cell volume reduction) and mitigating defenses (e.g., antioxidants, chaperone proteins, upregulation of mitochondria-encoded genes, enzyme regulation, etc.) in order to identify adaptive strategies that defend and adapt mitochondria in animals that can be frozen for six months or more every year. A particular focus is placed on freeze-responsive genes in wood frogs that are encoded on the mitochondrial genome including ATP6/8, ND4 and 16S RNA. These were strongly up-regulated during whole body freezing (24 h at −2.5 °C) in the liver and brain but showed opposing responses to two component stresses: strong upregulation in response to anoxia but no response to dehydration stress. This indicates that freeze-responsive upregulation of mitochondria-encoded genes is triggered by declining oxygen and likely has an adaptive function in supporting cellular energetics under indeterminate lengths of whole body freezing.

Metabolic responses to dehydration by liver of the wood frog, Rana sylvatica

1994 ◽  
Vol 72 (8) ◽  
pp. 1420-1425 ◽  
Author(s):  
Thomas A. Churchill ◽  
Kenneth B. Storey
Keyword(s):  
Total Body Water ◽  
Energy Charge ◽  
Rana Sylvatica ◽  
Body Water ◽  
Whole Body ◽  
Metabolic Responses ◽  
Wood Frogs ◽  
Form Analysis ◽  
Glucose Synthesis ◽  
Total Body

The metabolic responses by the liver to the evaporative loss of up to 60% of total body water were quantified in spring-collected wood frogs, Rana sylvatica, a freeze-tolerant species. Dehydration stimulated rapid hyperglycemia, liver glucose levels rising 3.8-fold to 90 nmol/mg protein (9.9 μmol/g wet mass) by the time that 10% of total body water had been lost. Glucose accumulation occurred at the expense of liver glycogen reserves, which fell over the course of dehydration, and was supported by a 5.8-fold increase in the activity of glycogen phosphorylase a in the liver, made up of increases in both the total phosphorylase activity expressed and the percentage of the enzyme in the active form. Analysis of changes in the levels of glycolytic intermediates in the liver over the course of dehydration showed sharp increases in glucose-6-phosphate and fructose-6-phosphate during the period of active glucose synthesis but no change in the levels of fructose-1,6-bisphosphate or triose phosphates. This indicated that an inhibitory block on glycolysis at the phosphofructokinase reaction helped to promote the diversion of glycogenolysis into glucose export. When water loss exceeded 10%, cellular energetics were affected; ATP levels fell progressively between 25 and 60% dehydration, but a concomitant drop in the total adenylate pool held the energy charge stable at 0.7–0.8 up to 35% dehydration. At extreme dehydration (50 and 60%), metabolic indicators of hypoxia stress appeared in the liver: lactate accumulated and the energy charge fell. The data show that a primary response to whole-body dehydration in wood frogs is the activation of liver glucose synthesis and this suggests that the production of glucose as a cryoprotectant during freezing in this species is probably derived from a pre-existing amphibian volume-regulatory response to dehydration.

Variation in amount of surrounding forest habitat influences the initial orientation of juvenile amphibians emigrating from breeding ponds

2008 ◽  
Vol 86 (2) ◽  
pp. 141-146 ◽  
Author(s):  
Leroy J. Walston ◽  
Stephen J. Mullin
Keyword(s):  
Positive Relationship ◽  
Rana Sylvatica ◽  
Environmental Cues ◽  
Bufo Americanus ◽  
Forest Habitat ◽  
Amphibian Species ◽  
Wood Frogs ◽  
Juvenile Dispersal ◽  
Woodland Ponds ◽  
Emigration Behavior

Juvenile dispersal is important for the persistence of ​amphibian populations. Previous studies have observed nonrandom orientation in juvenile amphibians emigrating from breeding ponds; however, the environmental cues associated with these movements are not well understood. We examined the emigration behavior of recently metamorphosed juveniles of three pond-breeding amphibian species from three woodland ponds. We found that juvenile small-mouthed salamanders ( Ambystoma texanum (Matthes, 1855)), American toads ( Bufo americanus Holbrook, 1836), and wood frogs ( Rana sylvatica LeConte, 1825) exhibited nonrandom orientation upon exiting the breeding ponds. Furthermore, we found a positive relationship between captures of juvenile small-mouthed salamanders and wood frogs and width of the surrounding forest habitat, indicating that these species are selecting areas with broader forested habitat upon exiting the breeding ponds. Our results indicate that migrating juvenile amphibians may rely on direct environmental cues because the orientation of small-mouthed salamanders and wood frogs was influenced by width of the surrounding forested habitat. These observations support previous studies suggesting that maintaining forest habitat, along at least a portion of breeding ponds, is important for the persistence of amphibian populations.

scholarly journals Effects of Road De-Icing Salts on Two Amphibian Species, Rana Sylvatica and Rana Pipiens, and Their Trematode Parasites

2021 ◽  
Author(s):  
Dino Milotic
Keyword(s):  
Rana Sylvatica ◽  
Amphibian Declines ◽  
High Salt ◽  
Amphibian Species ◽  
Wood Frogs ◽  
Leopard Frogs ◽  
Larval Amphibians ◽  
Potential Contaminant ◽  
Northern Leopard Frogs ◽  
Host Parasite

With ongoing amphibian declines, it is essential to determine possible contributors such as diseases and environmental contaminants that may increase susceptibility. A potential contaminant is road salt (mainly NaCl), which leaches into aquatic environments. I examined whether road salts make larval amphibians (tadpoles) more susceptible to trematode parasite infection, and also how these affect free-living trematode infectious stages (cercariae). I exposed Rana sylvatica (wood frogs) and R. pipiens (northern leopard frogs) to control, medium (400 mg/L), and high salt (800 mg/L) treatments, and then to trematodes. High salt tended to reduce wood frog anti-parasite behaviour and resistance to infection but the opposite was seen for R. pipiens, although these tadpoles had elevated lymphocyte counts in high salinity. Trematodes were differentially affected by increased salinities. The results suggest that host-parasite-environment interactions are complex, with species differentially affected by contaminants, which may lead to community shifts in predominant hosts and parasite species.

scholarly journals Activation of the Hippo Pathway in Rana sylvatica: Yapping Stops in Response to Anoxia

Life ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1422
Author(s):  
Aakriti Gupta ◽  
Kenneth B. Storey
Keyword(s):  
Dna Binding ◽  
Gene Transcription ◽  
Hippo Pathway ◽  
Rana Sylvatica ◽  
Whole Body ◽  
Nuclear Fraction ◽  
Wood Frogs ◽  
Downstream Targets ◽  
Protein Levels ◽  
The Hippo Pathway

Wood frogs (Rana sylvatica) display well-developed anoxia tolerance as one component of their capacity to endure prolonged whole-body freezing during the winter months. Under anoxic conditions, multiple cellular responses are triggered to efficiently cope with stress by suppressing gene transcription and promoting activation of mechanisms that support cell survival. Activation of the Hippo signaling pathway initiates a cascade of protein kinase reactions that end with phosphorylation of YAP protein. Multiple pathway components of the Hippo pathway were analyzed via immunoblotting, qPCR or DNA-binding ELISAs to assess the effects of 24 h anoxia and 4 h aerobic recovery, compared with controls, on liver and heart metabolism of wood frogs. Immunoblot results showed significant increases in the relative levels of multiple proteins of the Hippo pathway representing an overall activation of the pathway in both organs under anoxia stress. Upregulation of transcript levels further confirmed this. A decrease in YAP and TEAD protein levels in the nuclear fraction also indicated reduced translocation of these proteins. Decreased DNA-binding activity of TEAD at the promoter region also suggested repression of gene transcription of its downstream targets such as SOX2 and OCT4. Furthermore, changes in the protein levels of two downstream targets of TEAD, OCT4 and SOX2, established regulated transcriptional activity and could possibly be associated with the activation of the Hippo pathway. Increased levels of TAZ in anoxic hearts also suggested its involvement in the repair mechanism for damage caused to cardiac muscles during anoxia. In summary, this study provides the first insights into the role of the Hippo pathway in maintaining cellular homeostasis in response to anoxia in amphibians.

scholarly journals Effects of Road De-Icing Salts on Two Amphibian Species, Rana Sylvatica and Rana Pipiens, and Their Trematode Parasites

2021 ◽  
Author(s):  
Dino Milotic
Keyword(s):  
Rana Sylvatica ◽  
Amphibian Declines ◽  
High Salt ◽  
Amphibian Species ◽  
Wood Frogs ◽  
Leopard Frogs ◽  
Larval Amphibians ◽  
Potential Contaminant ◽  
Northern Leopard Frogs ◽  
Host Parasite

With ongoing amphibian declines, it is essential to determine possible contributors such as diseases and environmental contaminants that may increase susceptibility. A potential contaminant is road salt (mainly NaCl), which leaches into aquatic environments. I examined whether road salts make larval amphibians (tadpoles) more susceptible to trematode parasite infection, and also how these affect free-living trematode infectious stages (cercariae). I exposed Rana sylvatica (wood frogs) and R. pipiens (northern leopard frogs) to control, medium (400 mg/L), and high salt (800 mg/L) treatments, and then to trematodes. High salt tended to reduce wood frog anti-parasite behaviour and resistance to infection but the opposite was seen for R. pipiens, although these tadpoles had elevated lymphocyte counts in high salinity. Trematodes were differentially affected by increased salinities. The results suggest that host-parasite-environment interactions are complex, with species differentially affected by contaminants, which may lead to community shifts in predominant hosts and parasite species.

Dehydration tolerance in wood frogs: a new perspective on development of amphibian freeze tolerance

1993 ◽  
Vol 265 (6) ◽  
pp. R1324-R1332 ◽  
Author(s):  
T. A. Churchill ◽  
K. B. Storey
Keyword(s):  
Total Body Water ◽  
Protective Layer ◽  
Rana Sylvatica ◽  
Freeze Tolerance ◽  
Body Water ◽  
Wood Frogs ◽  
Glucose Levels ◽  
High Concentrations ◽  
Total Body ◽  
New Perspective

Wood frogs, Rana sylvatica, tolerate the loss of 50-60% of total body water during experimental dehydration. The rate of water loss for unprotected frogs is the same whether animals are frozen (at -2 degrees C) or unfrozen (at 1 degrees C) but is greatly reduced when frogs are frozen under a protective layer of moss. Dehydrational death could occur in as little as 7-9 days for unprotected animals; this indicates the importance for winter survival of selecting well-protected and damp hibernation sites. Prior dehydration affected the cooling and freezing properties of frogs, reducing supercooling point and the amount of ice formed after 24 h at -2 degrees C and acting synergistically with freezing exposure in stimulating cryoprotectant synthesis. Analysis of the effects of controlled dehydration at 5 degrees C showed that changes in body water content alone (without freezing) stimulated liver glycogenolysis and the export of high concentrations of glucose into blood and other organs. Autumn-collected frogs dehydrated to 50% of total body water lost showed glucose levels of 165-1,409 nmol/mg protein in different organs, increases of 9- to 313-fold compared with control values and reaching final levels very similar to those induced by freezing exposure. The data support the proposal that various adaptations for natural freeze tolerance may have been derived from preexisting mechanisms for dealing with water stress in amphibians and that cell volume change may be one of the signals involved in triggering and sustaining molecular adaptations (e.g., cryoprotectant output) that support freezing survival.

scholarly journals Effect of Bronchoscopic Lung Volume Reduction in Advanced Emphysema on Energy Balance Regulation

Respiration ◽  
2021 ◽  
pp. 1-8
Author(s):  
Karin Sanders ◽  
Karin Klooster ◽  
Lowie E.G.W. Vanfleteren ◽  
Guy Plasqui ◽  
Anne-Marie Dingemans ◽  
...  
Keyword(s):  
Energy Balance ◽  
Lung Volume ◽  
Respiratory Mechanics ◽  
Volume Reduction ◽  
Fat Free Mass ◽  
Whole Body ◽  
Energy Requirements ◽  
Lung Volume Reduction ◽  
Clinical Trial Registry ◽  
The Impact

<b><i>Background:</i></b> Hypermetabolism and muscle wasting frequently occur in patients with severe emphysema. Improving respiratory mechanics by bronchoscopic lung volume reduction (BLVR) might contribute to muscle maintenance by decreasing energy requirements and alleviating eating-related dyspnoea. <b><i>Objective:</i></b> The goal was to assess the impact of BLVR on energy balance regulation. <b><i>Design:</i></b> Twenty emphysematous subjects participated in a controlled clinical experiment before and 6 months after BLVR. Energy requirements were assessed: basal metabolic rate (BMR) by ventilated hood, total daily energy expenditure (TDEE) by doubly labelled water, whole body fat-free mass (FFM) by deuterium dilution, and physical activity by accelerometry. Oxygen saturation, breathing rate, and heart rate were monitored before, during, and after a standardized meal via pulse oximetry and dyspnoea was rated. <b><i>Results:</i></b> Sixteen patients completed follow-up, and among those, 10 patients exceeded the minimal clinically important difference of residual volume (RV) reduction. RV was reduced with median (range) 1,285 mL (−2,430, −540). Before BLVR, 90% of patients was FFM-depleted despite a normal BMI (24.3 ± 4.3 kg/m<sup>2</sup>). BMR was elevated by 130%. TDEE/BMR was 1.4 ± 0.2 despite a very low median (range) daily step count of 2,188 (739, 7,110). Following BLVR, the components of energy metabolism did not change significantly after intervention compared to before intervention, but BLVR treatment decreased meal-related dyspnoea (4.1 vs. 1.7, <i>p</i> = 0.019). <b><i>Conclusions:</i></b> Impaired respiratory mechanics in hyperinflated emphysematous patients did not explain hypermetabolism. <b><i>Clinical Trial Registry Number:</i></b> NCT02500004 at www.clinicaltrial.gov.

Use of Continuous Blood Volume Monitoring to Detect Inadequately High Dry Weight

1996 ◽  
Vol 19 (7) ◽  
pp. 411-414 ◽  
Author(s):  
F. Lopot ◽  
P. Kotyk ◽  
J. Bláha ◽  
J. Forejt
Keyword(s):  
Blood Volume ◽  
Vena Cava ◽  
Extracellular Fluid ◽  
Dry Weight ◽  
Whole Body ◽  
The Status ◽  
Varying Length ◽  
Group 3 ◽  
Group 2 ◽  
Total Body

A continuous blood volume monitoring (CBVM) device (Inline Diagnostics, Riverdale, USA) was used to study response to prescribed ultrafiltration during haemodialysis (HD) in 66 stabilised HD patients. Fifty percent of patients showed the expected linear decrease in BV right from the beginning of HD (group 1), 32% exhibited no decrease at all (group 2), while eighteen percent formed the transient group 3 which showed a plateau of varying length after which a decrease occurred. The correct setting of dry weight was verified through evaluation of the ratio of extracellular fluid volume to total body water (VEC/TBW) in 26 patients by means of whole body multifrequency impedometry MFI (Xitron Tech., San Diego, USA) and through measurement of the Vena Cava Inferior diameter (VCID) pre and post HD (in 6 and 5 patients from groups 1 and 3 and from group 2, respectively). The mean VEC/TBW in groups 1 and 3 was 0.56 pre and 0.51 post HD as compared to 0.583 and 0.551 in group 2. VCID decreased on average by 14.1% in groups 1 and 3 but remained stable in group 2. Both findings thus confirmed inadequately high estimation of dry weight. Since CBVM is extremely easy to perform it can be used as a method of choice in detecting inadequately high prescribed dry weight. The status of the cardiovascular system must always be considered before final judgement is made.

scholarly journals Smaller organ mass with greater age, except for heart

2009 ◽  
Vol 106 (6) ◽  
pp. 1780-1784 ◽  
Author(s):  
Qing He ◽  
Stanley Heshka ◽  
Jeanine Albu ◽  
Lawrence Boxt ◽  
Norman Krasnow ◽  
...  
Keyword(s):  
African Americans ◽  
The Elderly ◽  
Left Ventricular ◽  
Fat Free Mass ◽  
Whole Body ◽  
Organ Mass ◽  
Total Body ◽  
Age Range ◽  
Negative Relationships

Autopsy/cadaver data indicate that many organs and tissues are smaller in the elderly compared with young adults; however, in vivo data are lacking. The aim of this study was to determine whether the mass of specific high-metabolic-rate organs is different with increasing age, using MRI. Seventy-five healthy women (41 African-Americans and 34 Caucasians, age range 19–88 yr) and 36 men (8 African-Americans and 28 Caucasians, age range 19–84 yr) were studied. MRI-derived in vivo measures of brain, heart, kidneys, liver, and spleen were acquired. Left ventricular mass (LVM) was measured by either echocardiography or cardiac gated MRI. Total body fat mass and fat-free mass (FFM) were measured with a whole body dual-energy X-ray absorptiometry (DXA) scanner. Multiple regression analysis was used to investigate the association between the organ mass and age after adjustment for weight and height (or DXA measures of FFM), race, sex, and interactions among these variable. No statistically significant interaction was found among age, sex, and race in any regression model. Significant negative relationships between organ mass and age were found for brain ( P < 0.0001), kidneys ( P = 0.01), liver ( P = 0.001), and spleen ( P < 0.0001). A positive relationship between LVM and age was found after adjustment for FFM ( P = 0.037). These findings demonstrate that age has a significant effect on brain, kidneys, liver, spleen, and heart mass. The age effect was independent of race and sex.

Glucose-6-phosphate dehydrogenase deficiency and cardiac surgery

Perfusion ◽  
2010 ◽  
Vol 25 (6) ◽  
pp. 417-421 ◽  
Author(s):  
N. Dogra ◽  
GD Puri ◽  
SS Rana
Keyword(s):  
Cardiac Surgery ◽  
Blood Cells ◽  
Perioperative Management ◽  
Dehydrogenase Deficiency ◽  
Ischemia Reperfusion ◽  
Anaesthetic Management ◽  
Whole Body ◽  
Free Radical Production ◽  
Radical Production ◽  
Glucose 6 Phosphate Dehydrogenase

Cardiac surgery involving cardiopulmonary bypass (CPB) in its conventional form involves many processes leading to free radical production, such as perioperative ischemia, reperfusion, circulation of whole body blood through the CPB circuit, hypothermia and acidosis. The red blood cells of a glucose-6-phosphate dehydrogenase (G6PD)-deficient person are unable to scavenge these free radicals, resulting in haemolysis. Here, we describe the successful anaesthetic management of two G6PD-deficient children who underwent cardiac surgery, on and off CPB, without any obvious haemolytic reaction, followed by a discussion of the disorder, with specific consideration of perioperative management of such cases.

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