Limits to sustained energy intake. XXIII. Does heat dissipation capacity limit the energy budget of lactating bank voles?

Acknowledgements We are grateful to our technicians and several students for their help during this study and for animal care. We thank Catherine Hambly and Peter Thompson for technical assistance for the isotope analysis for the DLW measurements. We thank Ulf Bauchinger for stimulating discussion and his comments, and two anonymous referees for comments on the manuscript. Funding This project was supported by grants from the Polish Ministry of Science and Higher Education [0595/B/P01/2011/40 to E.T.S. and 8167/B/P01/2011/40 to P.K.], and Jagiellonian University [DS/WBINOZ/INOS/757 to P.K.].Peer reviewedPublisher PD

Is Experimental Evolution of an Increased Aerobic Exercise Performance in Bank Voles Mediated by Endocannabinoid Signaling Pathway?

The level of physical activity achieved in a given situation depends on both physiological abilities and behavioral characteristics (motivation). We used a unique animal model to test a hypothesis that evolution of an increased aerobic exercise performance can be facilitated by evolution of motivation to undertake physical activity, mediated by brain endocannabinoid system. Bank voles (Myodes glareolus) from “aerobic” A lines selected for 22 generations for high swim-induced aerobic metabolism (VO2swim) achieved 65% higher “voluntary maximum” VO2swim than voles from unselected, “control” C lines. In C lines, VO2swim was 24% lower than the maximum forced-running aerobic metabolism (VO2run), while in A lines VO2swim and VO2run were practically the same. Thus, the selection changed both the aerobic capacity and motivation to exercise at the top performance level. We applied a pharmacological treatment manipulation to test a hypothesis that the endocannabinoid signaling pathway 1) affects the voles performance in the aerobic exercise trials, and 2) has been modified in the selection process. Administration of the CB1 receptor antagonist (Rimonabant) did not affect the level of metabolism, but administration of the endocannabinoid reuptake inhibitor (AM404) decreased VO2swim both in A and C lines (4%, p = 0.03) and tended to decrease VO2run (2%, p = 0.07). The significant effect of AM404 suggests the involvement of endocannabinoids in signaling pathways controlling the motivation to be active. However, the response to AM404 did not differ between A and C lines (interaction effect, p ≥ 0.29). Thus, the results did not provide a support to the hypothesis that modifications of endocannabinoid signaling have played a role in the evolution of increased aerobic exercise performance in our experimental evolution model system.Summary StatementThe results corroborated involvement of endocannabinoids in the regulation of physical activity, but did not support the hypothesis that modification of endocannabinoid signaling played a role in the evolution of increased aerobic exercise performance in our experimental evolution model

Concerted Phenotypic Flexibility of Avian Erythrocyte Size and Number in Response to Dietary Anthocyanin Supplementation

Background: Endurance flight impose substantial oxidative costs on the avian oxygen delivery system. In particular, the accumulation of irreversible damage in red blood cells can reduce the capacity of blood to transport oxygen and limit aerobic performance. Many songbirds consume large amounts of anthocyanin-rich fruit, which is hypothesized to reduce oxidative costs, enhance post-flight regeneration, and enable greater aerobic capacity. While their antioxidant benefits appear most straightforward, the effects of anthocyanins on blood composition remain so far unknown. We fed thirty hand-raised European starlings (Sturnus vulgaris) two semisynthetic diets (with or without anthocyanin supplement) and manipulated the extent of flight activity in a wind tunnel (daily flying or non-flying for over two weeks) to test for their interactive effects on functionally important haematological variables. Results: Supplemented birds had on average 15% more and 4% smaller red blood cells compared to non-supplemented individuals and these diet effects were independent of flight manipulation. Haemoglobin content was 7% higher in non-supplemented flying birds compared to non-flying birds, while similar haemoglobin content was observed among supplemented birds that were flown or not. Neither diet nor flight activity influenced haematocrit. Conclusion: The concerted adjustments suggest that supplementation generally improved antioxidant protection in blood, which could prevent the excess removal of cells from the bloodstream and may have several implications on the oxygen delivery system, including improved gas exchange and blood flow. The flexible haematological response to dietary anthocyanins may also suggest that free-ranging species preferentially consume anthocyanin-rich fruits for their natural blood doping, oxygen delivery-enhancement effects

A Shift in the Thermoregulatory Curve as a Result of Selection for High Activity-Related Aerobic Metabolism

According to the “aerobic capacity model,” endothermy in birds and mammals evolved as a result of natural selection favoring increased persistent locomotor activity, fuelled by aerobic metabolism. However, this also increased energy expenditure even during rest, with the lowest metabolic rates occurring in the thermoneutral zone (TNZ) and increasing at ambient temperatures (Ta) below and above this range, depicted by the thermoregulatory curve. In our experimental evolution system, four lines of bank voles (Myodes glareolus) have been selected for high swim-induced aerobic metabolism and four unselected lines have been maintained as a control. In addition to a 50% higher rate of oxygen consumption during swimming, the selected lines have also evolved a 7.3% higher mass-adjusted basal metabolic rate. Therefore, we asked whether voles from selected lines would also display a shift in the thermoregulatory curve and an increased body temperature (Tb) during exposure to high Ta. To test these hypotheses we measured the RMR and Tb of selected and control voles at Ta from 10 to 34°C. As expected, RMR within and around the TNZ was higher in selected lines. Further, the Tb of selected lines within the TNZ was greater than the Tb of control lines, particularly at the maximum measured Ta of 34°C, suggesting that selected voles are more prone to hyperthermia. Interestingly, our results revealed that while the slope of the thermoregulatory curve below the lower critical temperature (LCT) is significantly lower in the selected lines, the LCT (26.1°C) does not differ. Importantly, selected voles also evolved a higher maximum thermogenesis, but thermal conductance did not increase. As a consequence, the minimum tolerated temperature, calculated from an extrapolation of the thermoregulatory curve, is 8.4°C lower in selected (−28.6°C) than in control lines (−20.2°C). Thus, selection for high aerobic exercise performance, even though operating under thermally neutral conditions, has resulted in the evolution of increased cold tolerance, which, under natural conditions, could allow voles to inhabit colder environments. Further, the results of the current experiment support the assumptions of the aerobic capacity model of the evolution of endothermy

Data from: Lower haematocrit, haemoglobin and red blood cell number in zebra finches acclimated to cold compared to thermoneutral temperature

Thermoregulation constitutes an important share of the energy budget of endotherms. Elevated thermoregulatory requirements must be met by oxygen supply through the blood, as heat is produced mainly via aerobic processes. In contrast to mammal studies, it remains unclear whether elevated thermoregulatory needs are followed by changes in haematological variables in birds. We investigated haematocrit (HCT), haemoglobin content per volume of blood (HGB), number of red blood cells (RBC), and size of the erythrocytes (RBC) in zebra finches (Taeniopygia guttata) acclimated to either cold or thermoneutral ambient temperatures under laboratory conditions. Seventy-nine females were maintained for six weeks either in cold (T = + 12 °C) or thermoneutral (T = + 32 °C) ambient temperature prior to blood collection. On average, HGB, HCT and RBC were significantly lower by about 10% in cold acclimated compared to thermoneutral acclimated birds. Only RBC was not different between the two acclimation temperatures. Mean HCT, one of the most commonly measured haematological variable for example was 53 ± 0.9 % (LSM ± s.e.m) in thermoneutral and 49 ± 0.8 % (LSM ± s.e.m) in cold acclimated zebra finches. On first sight, the observed lower values for three out of the four determined haematological variables in response to acclimation to cold question oxygen supply to be indeed a limiting factor for heat production. However, higher demands of oxygen supply due to increased thermoregulation in birds may instead require specific optimisation of blood viscosity and modulation by other cardiovascular properties. Nucleated red blood cells in birds may pose different strain on blood viscosity compared to non-nucleated mammalian erythrocytes and explain the contrasting response in haematological variables to temperature acclimation between birds and mammals

Haematological variables under temperature acclimation

Values of haematological variables collected from captive zebra finches acclimated either to cold or thermoneutral conditions. Data contains values of haematocrit, haemaglobin, red blood cell count, red blood cell area, body mass and food intake. Exel file

Selection for high activity-related aerobic metabolism does not alter the capacity of non-shivering thermogenesis in bank voles

An intriguing question is how the capacity of non-shivering thermogenesis (NST) - a special mechanism supporting endothermic thermoregulation in mammals - is affected by selection for high exercise metabolism. It has been proposed that high NST could be a mechanism to compensate for a low basal production of heat. On the other hand, high basal or activity metabolism is associated with physiological characteristics such as high performance of the circulatory system, which are also required for achieving a high NST. Here we tested whether selection for high aerobic exercise performance, which correlates with an increased basal metabolic rate, led to a correlated evolution of maximum and facultative NST. Therefore, we measured the NST of bank voles, 'Myodes (= Clethrionomys) glareolus', from lines selected for 13-14 generations (n = 46) for high aerobic metabolism achieved during swimming and from unselected, control lines (n = 46)

scripts

The set of scripts used for SNPs calling, expression analyses, Fst calculation and pedigree-based simulations

Genomic response to selection for predatory behavior in a mammalian model of adaptive radiation

If genetic architectures of various quantitative traits are similar, as studies on model organisms suggest, comparable selection pressures should produce similar molecular patterns for various traits. To test this prediction, we used a laboratory model of vertebrate adaptive radiation to investigate the genetic basis of the response to selection for predatory behavior and compare it with evolution of aerobic capacity reported in an earlier work. After 13 generations of selection, the proportion of bank voles (Myodes [=Clethrionomys] glareolus) showing predatory behavior was five times higher in selected lines than in controls. We analyzed the hippocampus and liver transcriptomes and found repeatable changes in allele frequencies and gene expression. Genes with the largest differences between predatory and control lines are associated with hunger, aggression, biological rhythms, and functioning of the nervous system. Evolution of predatory behavior could be meaningfully compared with evolution of high aerobic capacity, because the experiments and analyses were performed in the same methodological framework. The number of genes that changed expression was much smaller in predatory lines, and allele frequencies changed repeatably in predatory but not in aerobic lines. This suggests that more variants of smaller effects underlie variation in aerobic performance, whereas fewer variants of larger effects underlie variation in predatory behavior. Our results thus contradict the view that comparable selection pressures for different quantitative traits produce similar molecular patterns. Therefore, to gain knowledge about molecular-level response to selection for complex traits, we need to investigate not only multiple replicate populations but also multiple quantitative traits.This work was supported by the Polish Ministry of Science and Higher Education (N N303 816740 to P.K.) and Jagiellonian University (DS/WBINOZ/INOS/757 to P.K. and DS/WBINOZ/INOS/762 to W.B.). M.K. acknowledges support of the Spanish Ministry of Economy and Competitiveness, “Centro de Excelencia Severo Ochoa 2013-2017”, SEV-2012-020