Gene expression of sex steroid metabolizing enzymes and receptors in the skeletal muscle of migrant and resident subspecies of white-crowned sparrow (Zonotrichia leucophrys).

Circulating sex steroid concentrations vary dramatically across the year in seasonally breeding animals. The ability of circulating sex steroids to effect muscle function can be modulated by changes in intracellular expression of steroid metabolizing enzymes (e.g., 5α-reductase type 2 and aromatase) and receptors. Together, these combined changes in plasma hormones, metabolizing enzymes and receptors allow for seasonally appropriate changes in skeletal muscle function. We tested the hypothesis that gene expression of sex steroid metabolizing enzymes and receptors would vary seasonally in skeletal muscle and these changes would differ between a migrant and resident life history strategy. We quantified annual changes in plasma testosterone and gene expression in pectoralis and gastrocnemius skeletal muscles using quantitative polymerase chain reaction (qPCR) in free-living migrant (Zonotrichia leucophrys gambelii) and resident (Z. l. nuttalli) subspecies of white-crowned sparrow during breeding, pre-basic molt, and wintering life history stages. Pectoralis muscle profile was largest in migrants during breeding, while residents maintained large muscle profiles year-round. Circulating testosterone peaked during breeding in both subspecies. Pectoralis muscle androgen receptor mRNA expression was lower in females of both subspecies during breeding. Estrogen receptor-α expression was higher in the pectoralis muscle, but not gastrocnemius, of residents throughout the annual cycle when compared to migrants. Pectoralis aromatase expression was higher in resident males compared to migrant males. No differences were observed for 5α-reductase 2. Between these two subspecies, patterns of plasma testosterone and androgen receptors appear to be conserved, however estrogen receptor gene expression appears to have diverged.

A modified niche model for generating food webs with stage-structured consumers: The stabilizing effects of life-history stages on complex food webs.

Almost all organisms grow in size during their lifetime and switch diets, trophic positions, and interacting partners as they grow. Such ontogenetic development introduces life-history stages and flows of biomass between the stages through growth and reproduction. However, current research on complex food webs rarely considers life-history stages. The few previously proposed methods do not take full advantage of the existing food web structural models that can produce realistic food web topologies.We extended the niche model developed by Williams and Martinez (Nature, 2000, 404, 180-183) to generate food webs that included trophic species with a life-history stage structure. Our method aggregated trophic species based on niche overlap to form a life-history structured population; therefore, it largely preserved the topological structure of food webs generated by the niche model. We applied the theory of allometric predator-prey body mass ratio and parameterized an allometric bioenergetic model augmented with biomass flow between stages via growth and reproduction to study the effects of a stage structure on the stability of food webs.When life-history stages were linked via growth and reproduction, more food webs persisted, and persisting food webs tended to retain more trophic species. Topological differences between persisting linked and unlinked food webs were small to modest. The slopes of biomass spectra were lower, and weak interaction links were more prevalent in the linked food webs than the unlinked ones, suggesting that a life-history stage structure promotes characteristics that can enhance stability of complex food webs.Our results suggest a positive relationship between the complexity and stability of complex food webs. A life-history stage structure in food webs may play important roles in dynamics of and diversity in food webs.

Reactive scope model and emergency life history stage provide useful tools for evaluating the stress responses of native Australian lizards living in disturbed landscapes.

Glucocorticoids (GCs) are used as biomarkers of physiological stress response in reptiles. Fundamental stress physiology tools including the emergency life history stage (ELHS) and the reactive scope model (RSM) can be useful to determine how individual variation of stress responses shape population ecology. In this perspective, we applied the RSM and ELHS into the context of two urban-dwelling small native Australian reptile species to compare the stress-response patterns in short- and long-breeding lizards. Firstly, by drawing inferences from the ELHS, we presented hypothetical scenarios using sample GC data for a short-breeding species (e.g. common blue-tongue lizard). We showed that activation of the physiological stress response would be non-adaptive due to the consequences of stress on reproduction. Therefore, blue-tongue lizards may become exposed to acute and chronic environmental stressors (e.g. human disturbance and habitat clearance) during the breeding season as they prefer not to activate their hypothalamo-pituitary interrenal (HPI) axis in support of their short-breeding season. On the contrary, long-breeding lizards (e.g. bearded dragons), which have multiple breeding opportunities and are accustomed to living around humans and altered urban environments, tended to operate above the critical adaptive value of the ELHS during their breeding period. This suggests that any future changes to the dynamics of habitat availability and breeding opportunities may favour the dragon differently over the blue-tongue lizard. To further capture the dynamics of stress responses along spatial and temporal scales, we suggested that researchers should collect field data (e.g. blood plasma or faecal GCs) and then use the ELHS and RSM to understand how the environment is shaping the animal's stress physiology. The application of field stress monitoring and data visualization using the ELHS and RSM could guide environmental monitoring and conservation programs of native wildlife species.

Annual regulation of adrenocortical function in migrant and resident subspecies of white-crowned sparrow.

Corticosterone affects physiology and behavior both during normal daily processes but also in response to environmental challenges and is known to mediate life history trade-offs. Many studies have investigated patterns of corticosterone production at targeted times of year, while ignoring underlying annual profiles. We aimed to understand the annual regulation of hypothalamic-pituitary-adrenal (HPA) axis function of both migrant (Zonotrichia leucophrys gambelii; n = 926) and resident (Z. l. nutalli; n = 688) subspecies of white-crowned sparrow and how it is influenced by environmental conditions - wind, precipitation, and temperature. We predicted that more dramatic seasonal changes in baseline and stress-induced corticosterone would occur in migrants to precisely time the onset of breeding and cope with environmental extremes on their arctic breeding grounds, while changes in residents would be muted as they experience a more forgiving breeding schedule and comparatively benign environmental conditions in coastal California. During the course of a year, the harshest conditions were experienced the summer breeding grounds for migrants, at which point they had higher corticosterone levels compared to residents. For residents, the winter months coincided with harshest conditions at which point they had higher corticosterone levels than migrants. For both subspecies, corticosterone tended to rise as environmental conditions became colder and windier. We found that the annual maxima in stress-induced corticosterone occurred prior to egg lay for all birds except resident females. Migrants had much higher baseline and acute stress-induced corticosterone during breeding compared to residents; where in a harsher environment the timing of the onset of reproduction is more critical because the breeding season is shorter. Interestingly, molt was the only stage within the annual cycle in which subspecies differences were absent suggesting that a requisite reduction in corticosterone may have to be met for feather growth. These data suggest that modulation of the HPA axis is largely driven by environmental factors, social cues, and their potential interactions with a genetic program.

N-P utilization of Acer mono leaves at different life history stages across altitudinal gradients.

The relationship between plants and the environment is a core area of research in ecology. Owing to differences in plant sensitivity to the environment at different life history stages, the adaptive strategies of plants are a cumulative result of both their life history and environment. Previous research on plant adaptation strategies has focused on adult plants, neglecting saplings or seedlings, which are more sensitive to the environment and largely affect the growth strategy of subsequent life stages. We compared leaf N and P stoichiometric traits of the seedlings, saplings, and adult trees of Acer mono Maxim and different altitudes and found significant linear trends for both life history stages and altitude. Leaf N and P content by unit mass were greatly affected by environmental change, and the leaf N and P content by unit area varied greatly by life history stage. Acer mono leaf N-P utilization showed a significant allometric growth trend in all life history stages and at low altitudes. The adult stage had higher N-use efficiency than the seedling stage and exhibited an isometric growth trend at high altitudes. The N-P utilization strategies of A. mono leaves are affected by changing environmental conditions, but their response is further dependent upon the life history stage of the plant. Thus, this study provides novel insights into the nutrient use strategies of A. mono and how they respond to the environmental temperature, soil moisture content along altitude and how these changes differ among different life history stages, which further provide the scientific basis for the study of plant nutrient utilization strategy on regional scale.

Quantitative analysis of age and life-history stage related changes in DCX expression in the male Japanese quail (Cortunix japonica) telencephalon.

Most avian neurogenesis studies focused on the song control system and little attention has been given to non-song birds such as the Japanese quail. However, the only few neurogenesis studies in quails mainly focused on the sex steroid sensitive areas of the brain such as the medial preoptic and lateral septal nuclei. Despite the important role the quail telencephalon plays in filial imprinting and passive avoidance learning, neurogenesis in this structure has been completely overlooked. The aim of this study was therefore to quantitatively determine how DCX expression in the Japanese quail telencephalon changes with post hatching age (3-12 weeks) and life history stage. In this study, DCX was used as a proxy for neuronal incorporation. Bipolar and multipolar DCX immunoreactive cells were observed in the entire telencephalon except for the entopallium and arcopallium. In addition, DCX expression in all the eight telencephalic areas quantified was strongly negatively correlated with post-hatching age. Furthermore, numbers of bipolar and multipolar DCX immunoreactive cells were higher in the juvenile compared to subadult and adult quails. In conclusion, neuronal incorporation in the quail telencephalon is widespread but it declines with post hatching age. In addition, the most dramatic decline in neuronal incorporation in the telencephalic areas quantified takes place just after the birds have attained sexual maturity.

Age-related changes in Ki-67 and DCX expression in the BALB/ c mouse (Mus Musculus) brain.

Several studies have identified age as one of the strongest regulators of neurogenesis in the mammalian brain. However, previous age-related studies focused mainly on changes in neurogenesis during different stages of adulthood and did not describe changes in neurogenesis through the different life history stages of the animal. The aim of this study was therefore to determine time course changes in neurogenesis in the male BALB/c mouse brain at postnatal ages 1 week to 12 weeks, spanning juvenile, sub adult and adult life history stages. To achieve this, Ki-67 and DCX immunohistochemistry was used to assess changes in cell proliferation and neuronal incorporation respectively. Ki-67 expression was mainly observed in the olfactory bulb, rostral migratory stream, sub ventricular zone of lateral ventricle and the sub granular zone of the dentate gyrus. In addition, fewer Ki-67 positive cells were also observed in the neocortex, cerebellum and tectum. DCX was expressed in similar regions as Ki-67 except for the cerebellum and tectum. Expression of both Ki-67 and DCX sharply decreased with advancing age or life history stages in the sub ventricular zone, rostral migratory stream and sub granular zone of the BALB/c mouse brain. Neurogenesis therefore persists throughout all life history stages in the BALB/c mouse brain although it decreases with age.

Weathering the storm: Do arctic blizzards cause repeatable changes in stress physiology and body condition in breeding songbirds?

Severe weather events are increasing worldwide because of climate change. To cope with severe weather events, vertebrates rely on the stress response which is activated by the hypothalamic-pituitary adrenal (HPA) axis to adjust physiology and behavior. Previous studies have detailed changes in baseline concentrations of the stress hormone corticosterone during a single storm event, but little data exists on how stress physiology and body condition are adjusted as the storm progresses across multiple days. This represents a serious gap in our understanding of how birds respond physiologically over the duration of a storm. We documented arctic snowstorms that occurred over five consecutive years that were endured by Lapland longspurs (Calcarius lapponicus; 2012-2016) and in three consecutive years by white-crowned sparrows (Zonotrichia leucophrys gambelii; 2014-2016). Data were collected on storm-free days, during snowstorms ranging in length from 1 to 3 days, and the day immediately following a snowstorm. The specific aims were to understand how stress physiology, measured at baseline and in response to restraint handling, and body condition changed over multiple days of the storm, and if these responses were consistent across years. Snowstorms did not affect baseline corticosterone concentrations for either species except for female Lapland longspurs and male white-crowned sparrows in 2014. Lapland longspurs, regardless of sex, increased stress-induced (restraint handling) corticosterone in response to snowstorms in all years but 2013, which was characterized by unusually harsh conditions. Both sexes of White-crowned sparrows showed a significant increase in the stress-induced levels of corticosterone during snowstorms in one of the three years of the study. Stress-induced corticosterone concentrations were only different across each day of the storm in one year of the study for Lapland longspurs. Changes in fat and body mass were not uniform across years, but measurable increases in fat stores and body mass were detected in males of both species during the first day of a snowstorm with declines typically occurring by the second day. Our study showed that severe weather events often caused rapid increases in HPA axis activity and body condition, but these profiles are likely dependent upon ecological and environmental context within the breeding season.

Sexually Dimorphic Patterns of Cell Proliferation in the Brain Are Linked to Seasonal Life-History Transitions in Red-Sided Garter Snakes.

Seasonal rhythms in physiology and behavior are widespread across diverse taxonomic groups and may be mediated by seasonal changes in neurogenesis, including cell proliferation, migration, and differentiation. We examined if cell proliferation in the brain is associated with the seasonal life-history transition from spring breeding to migration and summer foraging in a free-ranging population of red-sided garter snakes (Thamnophis sirtalis) in Manitoba, Canada. We used the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) to label newly proliferated cells within the brain of adult snakes collected from the den during the mating season or from a road located along their migratory route. To assess rates of cell migration, we further categorized BrdU-labeled cells according to their location within the ventricular zone or parenchymal region of the nucleus sphericus (homolog of the amygdala), preoptic area/hypothalamus, septal nucleus, and cortex (homolog of the hippocampus). We found that cell proliferation and cell migration varied significantly with sex, the migratory status of snakes, and reproductive behavior in males. In most regions of interest, patterns of cell proliferation were sexually dimorphic, with males having significantly more BrdU-labeled cells than females prior to migration. However, during the initial stages of migration, females exhibited a significant increase in cell proliferation within the nucleus sphericus, hypothalamus, and septal nucleus, but not in any subregion of the cortex. In contrast, migrating males exhibited a significant increase in cell proliferation within the medial cortex but no other brain region. Because it is unlikely that the medial cortex plays a sexually dimorphic role in spatial memory during spring migration, we speculate that cell proliferation within the male medial cortex is associated with regulation of the hypothalamus-pituitary-adrenal axis. Finally, the only brain region where cell migration into the parenchymal region varied significantly with sex or migratory status was the hypothalamus. These results suggest that the migration of newly proliferated cells and/or the continued division of undifferentiated cells are activated earlier or to a greater extent in the hypothalamus. Our data suggest that sexually dimorphic changes in cell proliferation and cell migration in the adult brain may mediate sex differences in the timing of seasonal life-history transitions.

Rangifer management controls a climate-sensitive tundra state transition.

Rangifer (caribou/reindeer) management has been suggested to mitigate the temperature-driven transition of Arctic tundra into a shrubland state, yet how this happens is uncertain. Here we study this much focused ecosystem state transition in riparian areas, where palatable willows (Salix) are dominant tall shrubs and highly responsive to climate change. For the state transition to take place, small life stages must become tall and abundant. Therefore we predicted that the performance of small life stages (potential recruits) of the tall shrubs were instrumental to the focal transition, where Rangifer managed at high population density would keep the small-stage shrubs in a "browse trap" independent of summer temperature. We used a large-scale quasi-experimental study design that included real management units that spanned a wide range of Rangifer population densities and summer temperatures in order to assess the relative importance of these two driving variables. Ground surveys provided data on density and height of the small shrub life stages, while the distributional limit (shrubline) of established shrublands (the tall shrub life stage) was derived from aerial photographs. Where Rangifer densities were above a threshold of approximately 5 animals/km2 , we found, in accordance with the expectation of a "browse trap," that the small life stages of shrubs in grasslands were at low height and low abundance. At Rangifer densities below this threshold, the small life stages of shrubs were taller and more abundant indicating Rangifer were no longer in control of the grassland state. For the established shrubland state, we found that the shrubline was at a 100-m lower elevation in the management units where Rangifer had been browsing in summer as opposed to the migratory ranges with no browsing in summer. In both seasonal ranges, the shrubline increased 100 m per 1°C increment in temperature. Our study supports the proposal that Rangifer management within a sustainable range of animal densities can mitigate the much-focused transition from grassland to shrubland in a warming Arctic.

Effects of short-term fasting on stress physiology, body condition, and locomotor activity in wintering male white-crowned sparrows.

For wild free-living animals the availability of food resources can be greatly affected by environmental perturbations such as weather events. In response to environmental perturbations, animals activate the hypothalamic-pituitary-adrenal (HPA) axis to adjust physiology and behavior. The literature asserts that during weather events food intake declines leading to changes in HPA axis activity, as measured by both baseline and stress-induced glucocorticoid concentrations. Here we investigated how body condition, locomotor activity, and stress physiology were affected by varying lengths of a fast (1, 2, 6, and 24h; similar to that experienced by free-living birds) compared to when food was provided ad libitum in captive wintering male white-crowned sparrows, Zonotrichia leucophrys gambelii, exposed to a short day photoperiod. Baseline corticosterone concentrations were increased for all fasting durations but were highest in 6 and 24h fasted birds. Stress-induced corticosterone was elevated in 1h fasted birds with a trend for the 2h of fast; no other differences were found. Baseline corticosterone concentrations were negatively related to both total fat scores and body mass. All birds lost body mass regardless of fast length but birds fasted for 24h lost the most. Fat scores declined in the 6 and 24h groups, and no measureable changes were detected in pectoralis muscle profile. Locomotor activity was increased over the entire period in which food was removed regardless of fasting duration. Together this suggests that reduced food availability is responsible, at least in part, for the rapid elevation both baseline corticosterone under any duration of fast and stress-induced concentrations during short-term fasts.

Entanglement is a costly life-history stage in large whales.

Individuals store energy to balance deficits in natural cycles; however, unnatural events can also lead to unbalanced energy budgets. Entanglement in fishing gear is one example of an unnatural but relatively common circumstance that imposes energetic demands of a similar order of magnitude and duration of life-history events such as migration and pregnancy in large whales. We present two complementary bioenergetic approaches to estimate the energy associated with entanglement in North Atlantic right whales, and compare these estimates to the natural energetic life history of individual whales. Differences in measured blubber thicknesses and estimated blubber volumes between normal and entangled, emaciated whales indicate between 7.4 × 1010 J and 1.2 × 1011 J of energy are consumed during the course to death of a lethal entanglement. Increased thrust power requirements to overcome drag forces suggest that when entangled, whales require 3.95 × 109 to 4.08 × 1010 J more energy to swim. Individuals who died from their entanglements performed significantly more work (energy expenditure × time) than those that survived; entanglement duration is therefore critical in determining whales' survival. Significant sublethal energetic impacts also occur, especially in reproductive females. Drag from fishing gear contributes up to 8% of the 4-year female reproductive energy budget, delaying time of energetic equilibrium (to restore energy lost by a particular entanglement) for reproduction by months to years. In certain populations, chronic entanglement in fishing gear can be viewed as a costly unnatural life-history stage, rather than a rare or short-term incident.

Annual Hematocrit Profiles in Two Subspecies of White-Crowned Sparrow: A Migrant and a Resident Comparison.

Hematocrit is an easily measured parameter that can be used to assess changes in oxygen carrying capacity necessitated by fluctuations in metabolic demands. Most hematocrit studies draw conclusions from changes in hematocrit that occur over a small sampling interval without an understanding of the variation that exists across the annual cycle. White-crowned sparrows provide an excellent model system due to the existence of a resident subspecies (Zonotrichia leucophrys nuttalli) that serves as a natural control for a migrant subspecies (Zonotrichia leucophrys gambelii). Comparing these two subspecies allows for the investigation of adaptive physiological changes at each life-history stage (i.e., migration, breeding, molt, etc.) in response to changing metabolic demands. Of particular interest, this subspecies comparison, by both calendar month and life-history stage, allows for the separation of adaptive increases in hematocrit due to migration from the natural seasonal variation in hematocrit. Hematocrit levels for males and females ranged throughout the year between 42%-47% and 40%-47% in the resident and between 45%-58% and 45%-56% in the migrant. In both subspecies, hematocrit levels were elevated during the breeding season compared to the nonbreeding season, and levels were reduced in females during egg laying. When grouped by life-history stage, hematocrit levels were always higher in the migrant compared to the resident. During the months in which migration occurred, hematocrit levels were 10%-12% higher in the migrant compared to the resident subspecies. These data suggest differential regulation of hematocrit between the two subspecies that may be attributed to phenotypic plasticity or genetic differences.

Low female stress hormone levels are predicted by same- or opposite-sex sociality depending on season in wild Assamese macaques.

The social environment can have a powerful impact on an individual's stress response and thus affect health and biological fitness. Positive social interactions are particularly important for females of species living in complex societies, e.g. humans and non-human primates. Existing studies have mainly focussed on the effect of same-sex social interaction on the stress response, rather than both same- and opposite-sex social interaction simultaneously. However, consideration of both may be crucial since females may have different 'social needs' across different life-history stages. Applying the conceptual framework of allostasis, we tested the hypothesis that female allostatic load (measured through faecal glucocorticoid levels [fGCs]), of wild seasonally breeding Assamese macaques (Macaca assamensis), would increase if their social needs were not maintained in accordance with season. We found significant seasonal differences in same- and opposite-sex sociality which, depending on season, predicted female fGCs. In the mating season, females which spent more time close to males and more frequently groomed with them exhibited lower fGCs. In the non-mating season, when female-male interaction was infrequent, positive female-female sociality predicted lower fGCs. Our results support the hypothesis that same- and opposite-sex sociopositive interactions, specific to certain life-history stages, can mediate fGCs. We interpret this as a consequence of the positive direct and/or indirect effects of social contact in accordance with interactions pertaining to a given life-history stage, which are likely to impact positively upon fitness.

Polysaccharide structure of tetrasporic red seaweed Tichocarpus crinitus.

Sulfated polysaccharide isolated from tetrasporic plants of Tichocarpus crinitus was investigated. The polysaccharide was isolated by two methods: with water extraction at 80 °C (HT) and with a mild alkaline extraction (AE). The extracted polysaccharides were presented by non-gelling ones only, while galactose and 3,6-AG were the main monosaccharides, at the same time amount of 3,6-AG in AE polysaccharides was the similar to that of HT. According to methods of spectroscopy and mass spectrometry, the polysaccharide from tetrasporic T. crinitus contains main blocks of 1,3-linked ß-D-galactopyranosyl-2,4-disulfates and 1,4-linked 3,6-anhydro-α-D-galactopyranosyl while 6-sulfated 4-linked galactopyranosyl resudies are randomly distributed along the polysaccharide chain. The alkaline treatment of HT polysaccharide results in obtaining polysaccharide with regular structure that composed of alternating 1,3-linked ß-D-galactopyranosyl-2,4-disulfates and 1,4-linked 3,6-anhydro-α-D-galactopyranosyl residues. Native polysaccharide (HT) possessed both high anticoagulant and antiplatelet activity measured by fibrin clotting and platelet aggregation induced by collagen. This activity could be connected with peculiar chemical structure of HT polysaccharide which has high sulfation degree and contains also 3,6-anhydrogalactose in the polymer chain.

SELECTION OF INTERNAL CONTROL GENE FOR EXPRESSION STUDIES IN PORPHYRA HAITANENSIS (RHODOPHYTA) AT DIFFERENT LIFE-HISTORY STAGES(1).

Accurate gene quantification depends on the use of an appropriate internal control gene, which should be verified before its use for normalizing data. Housekeeping genes, which are expressed at relatively constant levels, are generally regarded as candidate internal control genes. To determine the ideal internal control for gene expression profiles for Porphyra haitanensis T. J. Chang et B. F. Zheng (Bangiales, Rhodophyta) at different life-history stages, we used absolute quantification to assess the expression levels of six housekeeping genes (18S ribosomal RNA, 30S ribosomal protein, glyceraldehyde-3-phosphate dehydrogenase, elongation factor 3, alpha-tubulin, and beta-tubulin) at the sporophyte and gametophyte stages. Housekeeping genes were selected by comparing the differences of observed copy numbers in sporophytes and in gametophytes. TubB (beta-tubulin) was found to be the optimal internal control gene, because it showed the smallest difference of gene expression. Compared with TubB, other housekeeping genes had greater variation of expression to different degrees.