Guodong Han

The Impact of Yangtze River Discharge, Ocean Currents and Historical Events on the Biogeographic Pattern of Cellana toreuma along the China Coast

Aim Genetic data were used to measure the phylogeographic distribution of the limpet, Cellana toreuma along the China coast in order to acsertain impacts of historic events, ocean currents and especially freshwater discharge from the Yangtze River on the connectivity of intertidal species with limited larval dispersal capability. Methodology/Principal Findings Genetic variation in 15 populations of C. toreuma (nu200a=u200a418), ranging from the Yellow Sea (YS), East China Sea (ECS) and South China Sea (SCS), were determined from partial mitochondrial cytochrome c oxidase subunit I gene. Genetic diversity and divergence based on haplotype frequencies were analyzed using CONTRIB, and AMOVA was used to examine genetic population structure. Historic demographic expansions were evaluated from both neutrality tests and mismatch distribution tests. Among the 30 haplotypes identified, a dominant haplotype No. 1 (H1) existed in all the populations, and a relatively abundant private haplotype (H2) in YS. Pairwise FST values between YS and the other two groups were relatively high and the percentage of variation among groups was 10.9%. Conclusions The high nucleotide and gene diversity in the YS, with large pairwise genetic distances and relatively high percentages of variation among groups, suggests that this group was relatively isolated from ECS and SCS. This is likely driven by historic events, ocean currents, and demographic expansion. We propose that freshwater discharge from the Yangtze River, which may act as physical barrier limiting the southward dispersal of larvae from northern populations, is especially important in determining the separation of the YS group from the rest of the Chinese populations of C. toreuma.

Metabolic energy sensors (AMPK and SIRT1), protein carbonylation, and cardiac failure as biomarkers of thermal stress in an intertidal limpet: linking energetic allocation with environmental temperature during aerial emersion

SUMMARY The effects of heat stress on organisms are manifested at the levels of organ function, metabolic activity, protein stability and gene expression. Here, we examined effects of high temperature on the intertidal limpet Cellana toreuma to determine how the temperatures at which (1) organ failure (cardiac function), (2) irreversible protein damage (carbonylation) and (3) expression of genes encoding proteins involved in molecular chaperoning (hsp70 and hsp90) and metabolic regulation (ampk and sirt1) occur compare with field temperatures, which commonly exceed 30°C and can reach 46°C. Heart failure, indexed by the Arrhenius break temperature, occurred at 34.3°C. Protein carbonylation rose significantly at 38°C. Genes for heat shock proteins HSP70 (hsp70) and HSP90 (hsp90), for two subunits of AMP-activated protein kinase (AMPK) (ampkα and ampkβ) and for histone/protein deacetylase SIRT1 (sirt1) all showed increased expression at 30°C. Temperatures of maximal expression differed among genes, as did temperatures at which upregulation ceased. Expression patterns for ampk and sirt1 indicate that heat stress influenced cellular energy homeostasis; above ~30°C, upregulation of ATP-generating pathways is suggested by elevated expression of genes for ampk; an altered balance between reliance on carbohydrate and lipid fuels is indicated by changes in expression of sirt1. These results show that C. toreuma commonly experiences temperatures that induce expression of genes associated with the stress response (hsp70 and hsp90) and regulation of energy metabolism (ampk and sirt1). At high temperatures, there is likely to be a shift away from anabolic processes such as growth to catabolic processes, to provide energy for coping with stress-induced damage, notably to proteins.

The importance of thermal history: costs and benefits of heat exposure in a tropical, rocky shore oyster

ABSTRACT Although thermal performance is widely recognised to be pivotal in determining species’ distributions, assessment of this performance is often based on laboratory-acclimated individuals, neglecting their proximate thermal history. The thermal history of a species sums the evolutionary history and, importantly, the thermal events recently experienced by individuals, including short-term acclimation to environmental variations. Thermal history is perhaps of greatest importance for species inhabiting thermally challenging environments and therefore assumed to be living close to their thermal limits, such as in the tropics. To test the importance of thermal history, the responses of the tropical oyster Isognomon nucleus to short-term differences in thermal environments were investigated. Critical and lethal temperatures and oxygen consumption were improved in oysters that previously experienced elevated air temperatures, and were associated with an enhanced heat shock response, indicating that recent thermal history affects physiological performance as well as inducing short-term acclimation to acute conditions. These responses were, however, associated with trade-offs in feeding activity, with oysters that experienced elevated temperatures showing reduced energy gain. Recent thermal history, therefore, seems to rapidly invoke physiological mechanisms that enhance survival of short-term thermal challenge but also longer term climatic changes and consequently needs to be incorporated into assessments of species’ thermal performances. Summary: Thermal responses and organismal vulnerability to climate largely depend on the proximate thermal history of individuals as exposure to acute heat events can enhance survival and thermal performance.

Stress modulation of cellular metabolic sensors: interaction of stress from temperature and rainfall on the intertidal limpet Cellana toreuma

In the natural environment, organisms are exposed to large variations in physical conditions. Quantifying such physiological responses is, however, often performed in laboratory acclimation studies, in which usually only a single factor is varied. In contrast, field acclimatization may expose organisms to concurrent changes in several environmental variables. The interactions of these factors may have strong effects on organismal function. In particular, rare events that occur stochastically and have relatively short duration may have strong effects. The present experiments studied levels of expression of several genes associated with cellular stress and metabolic regulation in a field population of limpet Cellana toreuma that encountered a wide range of temperatures plus periodic rain events. Physiological responses to these variable conditions were quantified by measuring levels of mRNA of genes encoding heat‐shock proteins (Hsps) and metabolic sensors (AMPKs and Sirtuin 1). Our results reveal high ratios of individuals in upregulation group of stress‐related gene expression at high temperature and rainy days, indicating the occurrence of stress from both prevailing high summer temperatures and occasional rainfall during periods of emersion. At high temperature, stress due to exposure to rainfall may be more challenging than heat stress alone. The highly variable physiological performances of limpets in their natural habitats indicate the possible differences in capability for physiological regulation among individuals. Our results emphasize the importance of studies of field acclimatization in unravelling the effects of environmental change on organisms, notably in the context of multiple changes in abiotic factors that are accompanying global change.

Temperature relations of aerial and aquatic physiological performance in a mid‐intertidal limpet Cellana toreuma: adaptation to rapid changes in thermal stress during emersion

The physiological performance of a mid-intertidal limpet Cellana toreuma was determined to study the physiological adaptation of intertidal animals to rapid changes and extreme temperatures during emersion. The relationship between the Arrhenius breakpoint temperature (ABT) and in situ operative body temperature was studied to predict the possible impact of climate change on the species. The temperature coefficient (Q10) of emersed animals was higher than that of submersed animals and the ratio of aerial: aquatic heart rate rose with increasing temperature. The ABTs of submersed and emersed animals were 30.2 and 34.2°C, respectively. The heart rate and levels of molecular biomarkers (hsps, ampkα, ampkβ and sirt1 mRNA) were determined in 48 h simulated semi-diurnal tides. There were no obvious changes of heart rate and gene expression during the transition between emersion and submersion at room temperature, although expressions of hsp70 and hsp90 were induced significantly after thermal stress. These results indicate that C. toreuma can effectively utilize atmospheric oxygen, and the higher Q10 and ABT of emersed animals are adaptations to the rapid change and extreme thermal stress during emersion. However, the in situ operative body temperature frequently exceeds the aerial ABT of C. toreuma, indicating the occurrence of large-scale mortality of C. toreuma in summer, and this species should be sensitive to increasing temperature in the scenario of climate change.

Temporal patterns of cardiac performance and genes encoding heat shock proteins and metabolic sensors of an intertidal limpet Cellana toreuma during sublethal heat stress

Intertidal invertebrates develop effective physiological adaptations to cope with the rapidly changing thermal environment in the intertidal zone. In the present study, the temporal patterns of heart rate, protein carbonyl groups, and genes encoding heat shock proteins (hsp70 and hsp90) and metabolic sensors (ampkα, ampkβ and sirt1) were measured to study the effect of sublethal heat stress on the cardiac function, oxidative stress, heat shock response and cellular metabolism of an intertidal limpet Cellana toreuma. All the physiological parameters are sensitive to temperature and duration of heat stress. Spearman correlation analysis revealed that the correlations between heart rate and levels of heat shock proteins mRNA and metabolic sensors mRNA were statistically significant. These results further suggest that cardiac function plays crucial roles in cellular energy metabolism and heat shock responses. The significant increase of protein carbonyl groups at 34°C after 4h exposure indicated that the failure of cardiac function and the increase of anaerobic metabolism partly leads to the increase of protein carbonyl groups. Generally, the physiological responses to heat stress are sensitive to temperature and are energy-consumptive, as indicated by the upregulation of metabolic sensors mRNA. However, the upregulation of heat shock proteins and metabolic sensors at the post-transcriptional level and related functions need to be confirmed in further experiments.

Anaerobic metabolism and thermal tolerance: the importance of opine pathways on survival of a gastropod after cardiac dysfunction.

Organisms on rocky shores are frequently exposed to high temperatures, which cause impairment of cardiac function and retard cellular oxygen delivery. However, some gastropods can survive at several degrees Celsius higher than their Arrhenius break temperature of cardiac function (ABT), indicating the importance of anaerobic metabolism for their thermal tolerance. We measured the global molecular responses to heat stress in limpet Cellana toreuma using 454 GS-FLX to investigate the variations of genes involved in anaerobic metabolism at high temperatures. Next, the gene expression levels of 4 anaerobic enzymes and activity of alanopine dehydrogenase (AlDH), which is involved in opine pathway, were measured in response to elevated temperature. A total of 19 heat shock proteins (HSPs) were determined using real-time PCR at different temperatures. At high temperatures, the extensive upregulation of HSP genes was an effective but energetically expensive form of protection to prevent thermal damage. The upregulation of hypoxia-inducible factor 1 alpha mRNA indicated the condition of cellular hypoxia and the high gene expression and enzyme activity of AlDH suggested that opine pathway was the main anaerobic pathway. These results implied that anaerobic metabolism was enhanced to provide energy in the face of thermal stress. Our findings highlight the ecological significance of the anaerobic metabolism of gastropods to thermal adaptation. For predicting the ecological impact of global warming on the distribution of gastropods, the role of anaerobic pathways should be evaluated.

High thermal stress responses of Echinolittorina snails at their range edge predict population vulnerability to future warming

Populations at the edge of their species distribution ranges are typically living at the physiological extreme of the environmental conditions they can tolerate. As a species response to global change is likely to be largely determined by its physiological performance, subsequent changes in environmental conditions can profoundly influence populations at range edges, resulting in range extensions or retractions. To understand the differential physiological performance among populations at their distribution range edge and center, we measured levels of mRNA for heat shock protein 70 (hsp70) as an indicator of temperature sensitivity in two high-shore littorinid snails, Echinolittorina malaccana and E. radiata, between 1°N to 36°N along the NW Pacific coast. These Echinolittorina snails are extremely heat-tolerant and frequently experience environmental temperatures in excess of 55u202f°C when emersed. It was assumed that animals exhibiting high temperature sensitivity will synthesize higher levels of mRNA, which will thus lead to higher energetic costs for thermal defense. Populations showed significant geographic variation in temperature sensitivity along their range. Snails at the northern range edge of E. malaccana and southern range edge of E. radiata exhibited higher levels of hsp70 expression than individuals collected from populations at the center of their respective ranges. The high levels of hsp70 mRNA in populations at the edge of a species distribution range may serve as an adaptive response to locally stressful thermal environments, suggesting populations at the edge of their distribution range are potentially more sensitive to future global warming.